Antibody formulations having optimized aggregation and fragmentation profiles

[0001] This application claims the benefit under 35 U.S.C. § 119(e) of U.S. Provisional Patent Application No. 60/693,603, filed on Jun. 23, 2005, and U.S. Provisional Patent Application No. 60/699,614, filed on Jul. 15, 2005, each of which is incorporated by reference herein in its entirety.

[0002] The present invention provides methods of optimizing the production and purification of antibody formulations that immunospecifically bind to antigens of interest and are suitable for parenteral administration to a subject, which formulations exhibit increased stability due to reduced degradation and aggregation of the antibody component on long term storage. Such methods provide formulations that offer multiple advantages over formulations produced by non-optimized methods including less stringent or more readily available transportation/storage conditions, and less frequent dosing or smaller dosage amounts in the therapeutic, prophylactic and diagnostic use of such formulations. The invention further provides methods of utilizing the formulations of the present invention. In a specific embodiment, the invention provides methods of optimizing the production and purification of antibody formulations that immunospecifically bind to RSV antigens, which formulations exhibit increased stability due to reduced degradation and aggregation of the antibody component on long term storage. Such formulations may be used in the diagnostic, therapeutic or prophylactic treatment of RSV infections.

[0003] Respiratory infections are common infections of the upper respiratory tract (e.g., nose, ears, sinuses, and throat) and lower respiratory tract (e.g., trachea, bronchial tubes, and lungs). Symptoms of upper respiratory infection include runny or stuffy nose, irritability, restlessness, poor appetite, decreased activity level, coughing, and fever. Viral upper respiratory infections cause and/or are associated with sore throats, colds, croup, and the flu. Clinical manifestations of a lower respiratory infection include shallow coughing that produces sputum in the lungs, fever, and difficulty breathing.

[0004] Respiratory syncytial virus (RSV) is one of the leading causes of respiratory disease worldwide. In the United States, it is responsible for tens of thousands of hospitalizations and thousands of deaths per year (see Black, C. P., Resp. Care 2003 48(3):209-31 for a recent review of the biology and management of RSV). Infants and children are most at risk for serious RSV infections which migrate to the lower respiratory system, resulting in pneumonia or bronchiolitis. In fact, 80% of childhood bronchiolitis cases and 50% of infant pneumonias are attributable to RSV. The virus is so ubiquitous and highly contagious that almost all children have been infected by two years of age. Although infection does not produce lasting immunity, reinfections tend to be less severe so that in older children and healthy adults RSV manifests itself as a cold or flu-like illness affecting the upper and/or lower respiratory system, without progressing to serious lower respiratory tract involvement. However, RSV infections can become serious in elderly or immunocompromised adults. (Evans, A. S., eds., 1989, Viral Infections of Humans. Epidemiology and Control, 3^rd ed., Plenum Medical Book, New York at pages 525-544; Falsey, A. R., 1991, Infect. Control Hosp. Epidemiol. 12:602-608; and Garvie et al., 1980, Br. Med. J. 281:1253-1254; Hertz et al., 1989, Medicine 68:269-281).

[0005] At present, there is no vaccine against RSV, nor is there any effective treatment. Recent clinical data has failed to support the early promise of the antiviral agent ribavirin, which is the only drug approved for treatment of RSV infection (Black, C. P., Resp. Care 2003 48(3):209-31). Consequently, the American Academy of Pediatrics issued new guidelines suggesting that use of ribavirin be restricted to only the most severe cases (Committee on Infectious Disease, American Academy of Pediatrics. 1996. Pediatrics 97:137-140; Randolph, A. G., and E. E. Wang., 1996, Arch. Pediatr. Adolesc. Med. 150:942-947).

[0006] While a vaccine or effective treatment have proven elusive, some success has been achieved in the area of prevention for infants at high risk of serious upper and/or lower respiratory tract RSV infection. In particular, there are two immunoglobulin-based therapies approved to protect high-risk infants from serious lower respiratory tract RSV infection, RSV-IGIV (RSV-immunoglobulin intravenous, also known as RespiGam™) and palivizumab (SYNAGIS®). However, neither RSV-IGIV nor palivizumab has been approved for use other than as a prophylactic agent for lower respiratory tract RSV infections.

[0007] RSV is easily spread by physical contact with contaminated secretions. The virus can survive for at least half an hour on hands and for hours on countertops and used tissues. The highly contagious nature of RSV is evident from the risk factors associated with contracting serious infections. One of the greatest risk factors is hospitalization, where in some cases in excess of 50% of the staff on pediatric wards were found to be infected (Black, C. P., Resp. Care 2003 48(3):209-31). Up to 20% of these adult infections are asymptomatic but still produce substantial shedding of the virus. Other risk factors include attendance at day care centers, crowded living conditions, and the presence of school-age siblings in the home. Importantly, an agent that is effective at clearing the virus from the upper and/or lower respiratory tract is likely to be effective in preventing its transmission. Thus, one promising approach to preventing serious RSV infections is the development of therapies to clear or block the virus from the upper and/or lower respiratory tract.

[0008] Although RSV-IVIG and palivizumab represent significant advances in the prevention of lower respiratory tract RSV infections, neither has demonstrated efficacy at permissible doses against the virus in the upper respiratory tract. In fact, RSV-IVIG failed to clear nasal RSV when administered as a nasal spray in amounts that were effective to clear pulmonary RSV in every animal of the treatment group (Prince et al., U.S. Pat. No. 4,800,078, issued Jan. 24, 1989). The interperitoneal route of administration also failed to clear RSV from the upper respiratory tract with the same efficacy as the lower respiratory tract. It has recently been noted that the immune response elicited by upper respiratory tract infections differs from that induced by lower respiratory infections (van Benten I. J. et al., J. Med. Virol. 2003 October; 71(2):290-7). Thus, a need exists for the prevention and treatment of upper and/or lower respiratory tract RSV infections.

[0009] Otitis media is an infection or inflammation of the middle ear. This inflammation often begins when infections that cause sore throats, colds, or other respiratory or breathing problems spread to the middle ear. These can be viral or bacterial infections. RSV is the principal virus that has been correlated with otitis media. Seventy-five percent of children experience at least one episode of otitis media by their third birthday. Almost half of these children will have three or more ear infections during their first 3 years. It is estimated that medical costs and lost wages because of otitis media amount to $5 billion a year in the United States (Gates G A, 1996, Cost-effectiveness considerations in otitis media treatment. Otolaryngol Head Neck Sur. 114 (4): 525-530). Although otitis media is primarily a disease of infants and young children, it can also affect adults.

[0010] Otitis media not only causes severe pain but may result in serious complications if it is not treated. An untreated infection can travel from the middle ear to the nearby parts of the head, including the brain. Although the hearing loss caused by otitis media is usually temporary, untreated otitis media may lead to permanent hearing impairment. Persistent fluid in the middle ear and chronic otitis media can reduce a child's hearing at a time that is critical for speech and language development. Children who have early hearing impairment from frequent ear infections are likely to have speech and language disabilities.

[0011] Although many physicians recommend the use of antibiotics for the treatment of ear infections, antibiotic resistance has become an important problem in effective treatment of the disease. Further, new therapies are needed to prevent or treat viral infections that are associated with otitis media, particularly RSV.

[0012] About 12 million people in the U.S. have asthma and it is the leading cause of hospitalization for children. The Merck Manual of Diagnosis and Therapy (17th ed., 1999).

[0013] Asthma is an inflammatory disease of the lung that is characterized by airway hyperresponsiveness (“AHR”), bronchoconstriction (i.e., wheezing), eosinophilic inflammation, mucus hypersecretion, subepithelial fibrosis, and elevated IgE levels. Asthmatic attacks can be triggered by environmental triggers (e.g. acarids, insects, animals (e.g., cats, dogs, rabbits, mice, rats, hamsters, guinea pigs, mice, rats, and birds), fungi, air pollutants (e.g., tobacco smoke), irritant gases, fumes, vapors, aerosols, chemicals, or pollen), exercise, or cold air. The cause(s) of asthma is unknown. However, it has been speculated that family history of asthma (London et al., 2001, Epidemiology 12(5):577-83), early exposure to allergens, such as dust mites, tobacco smoke, and cockroaches (Melen et al., 2001, 56(7):646-52), and respiratory infections (Wenzel et al., 2002, Am J Med, 112(8):672-33 and Lin et al., 2001, J Microbiol Immuno Infect, 34(4):259-64), such as RSV, may increase the risk of developing asthma. A review of asthma, including risk factors, animal models, and inflammatory markers can be found in O'Byrne and Postma (1999), Am. J. Crit. Care. Med. 159:S41-S66, which is incorporated herein by reference in its entirety.

[0014] Current therapies are mainly aimed at managing asthma and include the administration of β-adrenergic drugs (e.g. epinephrine and isoproterenol), theophylline, anticholinergic drugs (e.g., atropine and ipratorpium bromide), corticosteroids, and leukotriene inhibitors. These therapies are associated with side effects such as drug interactions, dry mouth, blurred vision, growth suppression in children, and osteoporosis in menopausal women. Cromolyn and nedocromil are administered prophylatically to inhibit mediator release from inflammatory cells, reduce airway hyperresponsiveness, and block responses to allergens. However, there are no current therapies available that prevent the development of asthma in subjects at increased risk of developing asthma. Thus, new therapies with fewer side effects and better prophylactic and/or therapeutic efficacy are needed for asthma.

[0015] Reactive airway disease is a broader (and often times synonymous) characterization for asthma-like symptoms, and is generally characterized by chronic cough, sputum production, wheezing or dyspenea.

[0016] Wheezing (also known as sibilant rhonchi) is generally characterized by a noise made by air flowing through narrowed breathing tubes, especially the smaller, tight airways located deep within the lung. It is a common symptom of RSV infection, and secondary RSV conditions such as asthma and brochiolitis. The clinical importance of wheezing is that it is an indicator of airway narrowing, and it may indicate difficulty breathing.

[0017] Wheezing is most obvious when exhaling (breathing out), but may be present during either inspiration (breathing in) or exhalation. Wheezing most often comes from the small bronchial tubes (breathing tubes deep in the chest), but it may originate if larger airways are obstructed.

[0018] Citation or discussion of a reference herein shall not be construed as an admission that such is prior art to the present invention.

[0019] The present invention is based upon the inventors' use of sensitive analytical techniques, such as analytical ultracentrifugation (AUC), size exclusion chromatography (SEC), Liquid Chromatography Mass Spectrometry (LC-MS) or particle counter analysis to analyze the fragmentation and aggregation profiles of formulations of full-length IgG1 monoclonal antibodies, particularly those that have been recombinantly expressed in myeloma cells, such as, but not limited to, NS0 cells. Thus, the present invention provides antibody formulations having fragmentation and aggregation profiles that are improved (i.e., have decreased total fragmentation and/or aggregation or have decreased amounts of certain types of fragments or aggregates or have reduced rates of aggregation or fragmentation) as compared to prior antibody formulations.

[0020] In a particular embodiment, the invention provides an antibody formulation comprising a full length IgG₁ antibody, preferably specific for a therapeutic or prophylactic target, wherein no more than 0.5% of the total protein fraction (in certain embodiments, however, at least 0.1% of the total protein fraction or is below detectable levels) of said formulation comprises (or, in other embodiments, consists of as impurities or as fragments to detectable levels) antibody type I fragments. In other embodiments, no more than 0.5% of the total protein fraction (and, in certain embodiments, at least 0.1% of the total protein fraction or is below detectable levels) of said formulation comprises (or, in other embodiments, consists of as impurities or as fragments to detectable levels) antibody type I fragments and antibody type II fragments. Preferably, the antibody type I fragments comprise one or more C-terminal portions of the heavy chain of the antibody, which heavy chain C-terminal portion has a molecular weight of about 25.6 kD, about 25.7 kD, about 25.8 kD, about 26.0 kD, or about 26.1 kD as determined by Liquid Chromatography Mass Spectrometry (LC-MS) analysis of samples of the antibody that have been deglycosylated, reduced and alkylated. Moreover, the antibody type II fragments comprise one or more N-terminal portions of the heavy chain of the antibody, which heavy chain N-terminal portion has a molecular weight of about 24.4 kD, about 24.6 kD, about 24.7 kD, about 24.9 kD, or about 25.1 kD as determined by LC-MS analysis of samples of the antibody that have been deglycosylated, reduced and alkylated. In addition, the antibody type I fragments may comprise one or more C-terminal portions of the heavy chain, which heavy chain C-terminal portion comprises amino acid residues 223-449 of the IgG1 heavy chain (according to Kabat numbering), amino acid residues 224-449 of the IgG1 heavy chain, amino acid residues 225-449 of the IgG1 heavy chain, amino acid residues 226-449 of the IgG1 heavy chain, amino acid residues 227-449 of the IgG1 heavy chain, amino acid residues 228-449 of the IgG1 heavy chain and amino acid residues 229-449 of the IgG1 heavy chain and the antibody type II fragments comprise one or more heavy chain N-terminal portions which comprises amino acid residues 1-222 of the IgG1 heavy chain, amino acid residues 1-223 of the IgG1 heavy chain, amino acid residues 1-224 of the IgG1 heavy chain, amino acid residues 1-225 of the IgG1 heavy chain, amino acid residues 1-226 of the IgG1 heavy chain, amino acid residues 1-227 of the IgG1 heavy chain or amino acid residues 1-228 of the IgG1 heavy chain. In certain embodiments, the antibody formulation does not contain detectable levels of any other types of fragments. In certain embodiments, the antibody formulation contains one, two, three, four, five, six or seven of the type I fragments and/or contains one, two, three, four, five, six or seven of the type II fragments.

[0021] In particular embodiments, the formulations of the invention comprise (or consists of as the aggregate fraction) a particle profile of less than about 3.4 E+5 particles/ml of diameter 2-4 μm, less than about 4.0 E+4 particles/ml of diameter 4-10 μm, less than about 4.2 E+3 particles/ml of diameter 10-20 μm, less than about 5.0 E+2 particles/ml of diameter 20-30 μm, less than about 7.5 E+1 particles/ml of diameter 30-40 μm, and less than about 9.4 particles/ml of diameter 40-60 μm as determined by a particle multisizer. In certain embodiments, the formulation contains no detectable particles greater than 40 μm, or greater than 30 μm. In other embodiments, the formulations of the invention have a turbidity value of a degassed sample of said formulation of about 6.4 NTU (in certain embodiments from 4-8 NTU, in other embodiments less than 10 NTU, less than 8 NTU, less than 7 NTU, or less than 6.5 NTU).

[0022] The antibody formulations of the invention may likewise have a combination of one or more of the above parameters of fragmentation and aggregation.

[0023] The antibody formulations of the invention are preferably at least 10 mg/ml antibody, more preferably, 15 mg/ml, 25 mg/ml, 50 mg/ml, 75 mg/ml, 100 mg/ml, 150 mg/ml or 200 mg/ml. The antibody in the antibody formulations of the invention may be any antibody that has a therapeutic, prophylactic or diagnostic utility. In preferred embodiments, the antibody in the formulations of the invention is specific for RSV and, in a specific embodiment, is not palivizumab. In more specific and preferred embodiments, the anti-RSV antigen binds to the F protein of RSV, and, in particular embodiments, the RSV antigen comprises or even consists of the F protein epitope NSELLSLINDMPITNDQKKLMSNN (SEQ ID NO:337). In other embodiments, the antibody is one of the antibodies listed in Table 2, preferably is the antibody A4B4L1FR-S28R or competes for binding with one of the antibodies listed in Table 2, preferably A4B4L1FR-S28R.

[0024] The antibody formulations of the invention preferably maintain improved aggregation and fragmentation profiles upon storage, for example, for extended periods (for example, but not limited to 6 months, 1 year, 2 years, 3 years or 5 years) at room temperature or 4° C. or for periods (such as, but not limited to 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 6 months or 1 year) at elevated temperatures such as 38° C.-42° C. Such formulations may be at pH 5-7, preferably at pH 6.0. Thus, in a particular embodiment, an antibody formulation of the invention comprising a full length IgG₁ antibody, upon storage at 38-42° C., pH 6.0 for 1 month, 6 months, 9 months or 14 months, comprises or, alternatively consists (other than the full length IgG1 antibody or as the fragment fraction) one or more antibody type I fragments. In another particular embodiment, an antibody formulation of the invention comprising a full length IgG₁ antibody, upon storage at 38-42° C., pH 6.0 for 1 month, 6 months, 9 months or 14 months, comprises or, alternatively consists (other than the full length IgG1 antibody or as the fragment fraction) one or more antibody type I fragments and one or more antibody type II fragments. Upon storage, the level of fragments as a percentage of the total amount of protein is preferably less than 0.5% and, in certain embodiments is at least 0.1% or is below detectable levels of fragments.

[0025] Additionally, during storage, such formulations preferably exhibit constant aggregation and fragmentation rates at temperatures, such as, but not limited to, 0-4° C., 10-15°, 20-24° C. room temperature, or elevated temperatures 38-42° C., and extended periods, such as, but not limited to, two weeks, one month, six months, one year, three years or five years. In certain embodiments, the antibody formulation does not contain detectable levels of any other types of fragments. Thus, in a particular embodiment, an antibody formulation of the invention comprising a full length IgG1 will increase in aggregate percentage relative to total protein, by 0.2%/month-0.35%/month at 20-24° C. and preferably by not more than 0.02%/month at 4° C. In a further embodiment, an antibody formulation of the invention comprising a full length IgG1 will not increase in fragment percentage, relative to total protein, by more than 0.015%/month-0.03%/month at 20-24° C. and preferably by not more than 0.00%/month at 4° C. In certain embodiments, the antibody formulation contains one, two, three, four, five or six or the type I fragments and/or contains one, two, three, four, five, six or seven of the type II fragments.

[0026] In particular embodiments, after storage, the formulations of the invention comprise (or consists of as the aggregate fraction) a particle profile of less than about 3.4 E+5 particles/ml of diameter 2-4 μm, less than about 4.0 E+4 particles/ml of diameter 4-10 μm, less than about 4.2 E+3 particles/ml of diameter 10-20 μm, less than about 5.0 E+2 particles/ml of diameter 20-30 μm, less than about 7.5 E+1 particles/ml of diameter 30-40 μm, and less than about 9.4 particles/ml of diameter 40-60 μm as determined by a particle multisizer. In certain embodiments, the formulation contains no detectable particles greater than 40 μm, or greater than 30 μm. In other embodiments, the formulations of the invention, after storage, have a turbidity value of a degassed sample of said formulation of about 6.4 NTU (in certain embodiments from 4-8 NTU, in other embodiments less than 10 NTU, less than 8 NTU, less than 7 NTU, or less than 6.5 NTU).

[0027] The antibody formulations of the invention, after storage, may likewise have a combination of one or more of the above parameters of fragmentation and aggregation.

[0028] Other aspects of the invention provide for methods of optimizing a particular antibody formulation for the fragmentation and aggregation parameters set forth above. Such methods comprise production, purification and formulation of the antibody and monitoring at one or more steps, or of the final formulation, for the levels of fragmentation and/or aggregation using methods such as, but not limited to AUC, SEC, LC-MS or particle multisizing, and then varying one or more parameters of one or more steps of the production, purification and/or formulation process or the formulation itself and evaluating whether varying the parameter reduces the level of fragmentation and/or aggregation. By such screening and monitoring steps, the method of the invention may be used to optimize an antibody formulation. Such parameters include, the temperature at which one or more steps is carried out, the reduction in or elimination of freeze/thaw cycles of the antibody, introduction of filtration steps, such as ultrafiltration, addition or change in one or more column chromatography steps, change in pH, etc.

[0029] The invention provides an antibody comprising a Fab fragment, which immunospecifically binds to an RSV antigen (e.g., the F protein epitope NSELLSLINDMPITNDQKKLMSNN (SEQ ID NO:337)), wherein the Tm of the Fab fragment is at least about 87° C., and wherein said antibody is not any of palivizumab, AFFF, P12f2, P12f4, P11d4, Ale9, A12a6, A13c4, A17d4, A4B4, A8c7, 1X-493L1FR, H3-3F4, M3H9, Y10H6, DG, AFFF(1), 6H8, L1-7E5, L2-15B10, A13a11, A1h5, A4B4(1), A4B4L1FR-S28R (motavizumab), A4B4-F52S, A17d4(1), A3e2, A14a4, A16b4, A17b5, A17f5, and A17h4. In a specific embodiment, the Fab in such an antibody is different from the Fab of palivizumab. In another embodiment, such an antibody comprises a VH or VL domain that is different from the VH or VL domain of palivizumab. In preferred embodiment, the Tm of the Fab fragment is at least about 90° C. or at least about 93° C. In another preferred embodiment, the pI of the antibody is between about 8.5 to 9.5 or between about 9.0 to 9.5.

[0030] In another specific embodiment, the antibody comprises a VH domain of the antibody A4B4L1FR-S28R (SEQ ID NO:48). In still another embodiment, the antibody comprises a VL domain of the antibody A4B4L1FR-S28R (SEQ ID NO:11). In still another embodiment, said Fab is the Fab of antibody A4B4L1FR-S28R.

[0031] The invention also provides an antibody formulation comprising the above described antibody, said formulation having a viscosity of less than 10.00 cP at any temperature in the range of 1 to 26° C.

[0032] The invention also provides an antibody formulation comprising the above described antibody, said formulation having an aggregration rate of less than 15% per day at any temperature in the range of 38 to 42° C.

[0033] The invention also provides a method of preventing, treating, or ameliorating one or more symptoms associated with a RSV infection in a subject, e.g., otitis media, asthma, and wheezing, said method comprising administering a prophylactically or therapeutically effective amount of an antibody formulation comprising such antibody. In one embodiment, the formulation is administered parenterally, intramuscularly, intravenously, subcutaneously or intranasally.

[0034] The invention also provides an antibody formulation comprising a full length IgG₁ antibody, which immunospecifically binds to an RSV antigen, said formulation having a viscosity of less than 10.00 cP at any temperature in the range of 1 to 26° C. The invention also provides an antibody formulation comprising any such antibody, said formulation having an aggregration rate of less than 15% per day at any temperature in the range of 38 to 42° C. In one embodiment, the antibody is not palivizumab. In another embodiment, the antibody is not any of AFFF, P12f2, P12f4, P11d4, Ale9, A12a6, A13c4, A17d4, A4B4, A8c7, 1X-493L1FR, H3-3F4, M3H9, Y10H6, DG, AFFF(1), 6H8, L1-7E5, L2-15B10, A13a11, A1h5, A4B4(1), A4B4L1FR-S28R (motavizumab), A4B4-F52S, A17d4(1), A3e2, A14a4, A16b4, A17b5, A17f5, and A17h4.

[0035] In the context of a polypeptide, the term “analog” as used herein refers to a polypeptide that possesses a similar or identical function as a RSV polypeptide, a fragment of a RSV polypeptide, or an antibody but does not necessarily comprise a similar or identical amino acid sequence of a RSV polypeptide, a fragment of a RSV polypeptide, or an antibody, or possess a similar or identical structure of a RSV polypeptide, a fragment of a RSV polypeptide, or an antibody. A polypeptide that has a similar amino acid sequence refers to a polypeptide that satisfies at least one of the following: (a) a polypeptide having an amino acid sequence that is at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or at least 99% identical to the amino acid sequence of a RSV polypeptide, a fragment of a RSV polypeptide, or an antibody described herein; (b) a polypeptide encoded by a nucleotide sequence that hybridizes under stringent conditions to a nucleotide sequence encoding a RSV polypeptide, a fragment of a RSV polypeptide, or an antibody described herein of at least 5 amino acid residues, at least 10 amino acid residues, at least 15 amino acid residues, at least 20 amino acid residues, at least 25 amino acid residues, at least 40 amino acid residues, at least 50 amino acid residues, at least 60 amino residues, at least 70 amino acid residues, at least 80 amino acid residues, at least 90 amino acid residues, at least 100 amino acid residues, at least 125 amino acid residues, or at least 150 amino acid residues; and (c) a polypeptide encoded by a nucleotide sequence that is at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or at least 99% identical to the nucleotide sequence encoding a RSV polypeptide, a fragment of a RSV polypeptide, or an antibody described herein. A polypeptide with similar structure to a RSV polypeptide, a fragment of a RSV polypeptide, or an antibody described herein refers to a polypeptide that has a similar secondary, tertiary or quaternary structure of a RSV polypeptide, a fragment of a RSV, or an antibody described herein. The structure of a polypeptide can determined by methods known to those skilled in the art, including but not limited to, X-ray crystallography, nuclear magnetic resonance, and crystallographic electron microscopy.

[0036] To determine the percent identity of two amino acid sequences or of two nucleic acid sequences, the sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in the sequence of a first amino acid or nucleic acid sequence for optimal alignment with a second amino acid or nucleic acid sequence). The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, then the molecules are identical at that position. The percent identity between the two sequences is a function of the number of identical positions shared by the sequences (i.e., % identity=number of identical overlapping positions/total number of positions×100%). In one embodiment, the two sequences are the same length.

[0037] The determination of percent identity between two sequences can also be accomplished using a mathematical algorithm. A preferred, non limiting example of a mathematical algorithm utilized for the comparison of two sequences is the algorithm of Karlin and Altschul, 1990, Proc. Natl. Acad. Sci. U.S.A. 87:2264 2268, modified as in Karlin and Altschul, 1993, Proc. Natl. Acad. Sci. U.S.A. 90:5873 5877. Such an algorithm is incorporated into the NBLAST and XBLAST programs of Altschul et al., 1990, J. Mol. Biol. 215:403. BLAST nucleotide searches can be performed with the NBLAST nucleotide program parameters set, e.g., for score=100, wordlength=12 to obtain nucleotide sequences homologous to a nucleic acid molecules of the present invention. BLAST protein searches can be performed with the XBLAST program parameters set, e.g., to score 50, wordlength=3 to obtain amino acid sequences homologous to a protein molecule of the present invention. To obtain gapped alignments for comparison purposes, Gapped BLAST can be utilized as described in Altschul et al., 1997, Nucleic Acids Res. 25:3389 3402. Alternatively, PSI BLAST can be used to perform an iterated search which detects distant relationships between molecules (Id.). When utilizing BLAST, Gapped BLAST, and PSI Blast programs, the default parameters of the respective programs (e.g., of XBLAST and NBLAST) can be used (see, e.g., http://www.ncbi.nlm.nih.gov). Another preferred, non limiting example of a mathematical algorithm utilized for the comparison of sequences is the algorithm of Myers and Miller, 1988, CABIOS 4:11 17. Such an algorithm is incorporated in the ALIGN program (version 2.0) which is part of the GCG sequence alignment software package. When utilizing the ALIGN program for comparing amino acid sequences, a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4 can be used.

[0038] The percent identity between two sequences can be determined using techniques similar to those described above, with or without allowing gaps. In calculating percent identity, typically only exact matches are counted.

[0039] The terms “antibodies that immunospecifically bind to a RSV antigen” and analogous terms as used herein refers to antibodies that specifically bind to a RSV polypeptide or a fragment of a RSV polypeptide and do not non-specifically bind to other polypeptides. Antibodies that immunospecifically bind to a RSV polypeptide or fragment thereof may have cross-reactivity with other antigens. Preferably, antibodies that immunospecifically bind to a RSV polypeptide or fragment thereof do not cross-react with other antigens. Antibodies that immunospecifically bind to a RSV polypeptide can be identified, for example, by immunoassays or other techniques known to those of skill in the art.

[0040] Antibodies of the invention include, but are not limited to, synthetic antibodies, monoclonal antibodies, recombinantly produced antibodies, multispecific antibodies (including bi-specific antibodies), human antibodies, humanized antibodies, chimeric antibodies, intrabodies, single-chain Fvs (scFv) (e.g., including monospecific and bi-specific, etc.), Fab fragments, F(ab′) fragments, disulfide-linked Fvs (sdFv), anti-idiotypic (anti-Id) antibodies, and epitope-binding fragments of any of the above. In particular, antibodies of the present invention include immunoglobulin molecules and immunologically active portions of immunoglobulin molecules, i.e., molecules that contain an antigen-binding site that immunospecifically binds to a RSV antigen (preferably, a RSV F antigen) (e.g., one or more complementarity determining regions (CDRs) of an anti-RSV antibody). The antibodies of the invention can be of any type (e.g., IgG, IgE, IgM, IgD, IgA and IgY), class (e.g., IgG₁, IgG₂, IgG₃, IgG₄, IgA₁, and IgA₂) or a subclass of immunoglobulin molecule.

[0041] As used herein, the term “analogue” in the context of a non-proteinaceous analog refers to a second organic or inorganic molecule which possess a similar or identical function as a first organic or inorganic molecule and is structurally similar to the first organic or inorganic molecule.

[0042] The term “antibody fragment” as used herein refers to a fragment of an antibody that immunospecifically binds to a RSV antigen. Antibody fragments may be generated by any technique known to one of skill in the art and by proteolytic or non-proteolytic cleavage. For example, Fab and F(ab′)₂ fragments may be produced by proteolytic cleavage of immunoglobulin molecules, using enzymes such as papain (to produce Fab fragments) or pepsin (to produce F(ab′)₂ fragments). F(ab′)₂ fragments contain the complete light chain, and the variable region, the CH1 region and the hinge region of the heavy chain. Antibody fragments can be also produced by recombinant DNA technologies. Antibody fragments may be one or more complementarity determining regions (CDRs) of antibodies.

[0043] The term “antibody type I fragment” as used herein refers to a multimeric protein comprising a full length antibody light chain, a full length antibody heavy chain and a C-terminal portion of an antibody heavy chain that, in human IgG₁ immunoglobulins, has an N-terminus at cysteine 223, aspartic acid 224, lysine 225, threonine 226, histidine 227, threonine 228 or cysteine 229 and a C-terminus at lysine 449. Amino acid numbering for the constant domain is given according to the Kabat EU numbering scheme (Kabat, E. A., T. T. Wu, H. M. Perry, K. S. Gottesman, and Foeller. 1991. Sequences of Proteins of Immunological Interest, U.S. Public Health Service, National Institutes of Health, Washington, D.C., which is incorporated herein by reference), unless otherwise indicated. In a specific embodiment, the full length antibody light chain, full length antibody heavy chain and C-terminal portion of an antibody heavy chain are linked by disulfide bonds as depicted in FIG. 14. In another specific embodiment, the type I fragment is capable of immunospecifically binding to an antigen of interest.

[0044] The term “antibody type II fragment” as used herein refers to a peptide, polypeptide, or protein comprising an antibody light chain and an N-terminal portion of an antibody heavy chain that, in human IgG₁ immunoglobulins, has a C-terminus at serine 222, cysteine 223, aspartic acid 224, lysine 225, threonine 226, histidine 227 or threonine 228 and an N-terminus at glycine 1. In a specific embodiment, the full length antibody light chain and N-terminal portion of an antibody heavy chain are linked by disulfide bonds as depicted in FIG. 14. In another specific embodiment, the type II fragment is capable of immunospecifically binding to an antigen of interest.

[0045] In the context of a polypeptide, the term “derivative” as used herein refers to a polypeptide that comprises an amino acid sequence of a RSV polypeptide, a fragment of a RSV polypeptide, or an antibody that immunospecifically binds to a RSV polypeptide which has been altered by the introduction of amino acid residue substitutions, deletions or additions. The term “derivative” as used herein also refers to a RSV polypeptide, a fragment of a RSV polypeptide, or an antibody that immunospecifically binds to a RSV polypeptide which has been modified, i.e., by the covalent attachment of any type of molecule to the polypeptide. For example, but not by way of limitation, a RSV polypeptide, a fragment of a RSV polypeptide, or an antibody may be modified, e.g., by glycosylation, acetylation, pegylation, phosphorylation, amidation, derivatization by known protecting/blocking groups, proteolytic cleavage, linkage to a cellular ligand or other protein, etc. A derivative of a RSV polypeptide, a fragment of a RSV polypeptide, or an antibody may be modified by chemical modifications using techniques known to those of skill in the art, including, but not limited to specific chemical cleavage, acetylation, formylation, metabolic synthesis of tunicamycin, etc. Further, a derivative of a RSV polypeptide, a fragment of a RSV polypeptide, or an antibody may contain one or more non-classical amino acids. A polypeptide derivative possesses a similar or identical function as a RSV polypeptide, a fragment of a RSV polypeptide, or an antibody described herein.

[0046] As used herein, the term “derivative” in the context of a non-proteinaceous derivative refers to a second organic or inorganic molecule that is formed based upon the structure of a first organic or inorganic molecule. A derivative of an organic molecule includes, but is not limited to, a molecule modified, e.g., by the addition or deletion of a hydroxyl, methyl, ethyl, carboxyl or amine group. An organic molecule may also be esterified, alkylated and/or phosphorylated.

[0047] The term “effective amount” as used herein refers to the amount of a therapy (e.g., an antibody of the invention) which is sufficient to reduce and/or ameliorate the severity and/or duration of a disease or disorder. For example, the “effective amount” of an anti RSV antibody is that which is sufficient to reduce and/or ameliorate the severity and/or duration of an upper and/or lower respiratory tract RSV infection, otitis media, and/or a symptom or respiratory condition relating thereto (including, but not limited to, asthma, wheezing, RAD, or a combination thereof), prevent the advancement or progression of the upper and/or lower respiratory tract RSV infection, otitis media and/or a symptom or respiratory condition relating thereto (e.g., prevent the progression of an upper respiratory tract RSV infection to a lower respiratory tract RSV infection), prevent the recurrence, development, or onset of an upper and/or lower respiratory tract RSV infection, otitis media, and/or a symptom or respiratory condition relating thereto (including, but not limited to, asthma, wheezing, RAD, or a combination thereof), and/or enhance/improve the prophylactic or therapeutic effect(s) of another therapy (e.g., a therapy other than an antibody of the invention). Non-limiting examples of effective amounts of an antibody of the invention are provided in Section 5.3, infra. With respect to the treatment of a RSV infection, a therapeutically effective amount refers to the amount of a therapeutic agent sufficient to reduce or inhibit the replication of a virus, inhibit or reduce the infection of cell with the virus, inhibit or reduce the production of the viral particles, inhibit or reduce the release of viral particles, inhibit or reduce the spread of the virus to other tissues or subjects, or ameliorate one or more symptoms associated with the infection. In a specific embodiment, a therapeutically effective amount of a therapeutic agent reduces one or more of the following steps of a RSV life cycle: the docking of the virus particle to a cell, the introduction of viral genetic information into a cell, the expression of viral proteins, the production of new virus particles and the release of virus particles from a cell by at least 5%, preferably at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 100%. In another specific embodiment, a therapeutically effective amount of a therapeutic agent reduces the replication, multiplication or spread of a virus by at least 5%, preferably at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 100%.

[0048] The term “effective neutralizing titer” of an anti-RSV antibody as used herein refers to the amount of antibody which corresponds to the amount present in the serum of animals (human or cotton rat) that has been shown to be either clinically efficacious (in humans) or to reduce virus by 99% in, for example, cotton rats. The 99% reduction is defined by a specific challenge of, e.g., 10³ pfu, 10⁴ pfu, 10⁵ pfu, 10⁶ pfu, 10⁷ pfu, 10⁸ pfu, or 10⁹ pfu of RSV.

[0049] The term “elderly” as used herein refers to a human subject who is age 65 or older.

[0050] The term “epitopes” as used herein refers to fragments of a polypeptide having antigenic or immunogenic activity in an animal, preferably a mammal, and most preferably in a human. An epitope having immunogenic activity is a fragment of a polypeptide that elicits an antibody response in an animal. An epitope having antigenic activity is a fragment of a polypeptide to which an antibody immunospecifically binds as determined by any method well known in the art, for example, by the immunoassays described herein. Antigenic epitopes need not necessarily be immunogenic.

[0051] The term “excipients” as used herein refers to inert substances which are commonly used as a diluent, vehicle, preservatives, binders, or stabilizing agent for drugs and includes, but not limited to, proteins (e.g., serum albumin, etc.), amino acids (e.g., aspartic acid, glutamic acid, lysine, arginine, glycine, histidine, etc.), fatty acids and phospholipids (e.g., alkyl sulfonates, caprylate, etc.), surfactants (e.g., SDS, polysorbate, nonionic surfactant, etc.), saccharides (e.g., sucrose, maltose, trehalose, etc.) and polyols (e.g., mannitol, sorbitol, etc.). Also see Remington's Pharmaceutical Sciences (by Joseph P. Remington, 18th ed., Mack Publishing Co., Easton, Pa.), which is hereby incorporated in its entirety.

[0052] The term “fragment” as used herein refers to a peptide or polypeptide comprising an amino acid sequence of at least 5 contiguous amino acid residues, at least 10 contiguous amino acid residues, at least 15 contiguous amino acid residues, at least 20 contiguous amino acid residues, at least 25 contiguous amino acid residues, at least 40 contiguous amino acid residues, at least 50 contiguous amino acid residues, at least 60 contiguous amino residues, at least 70 contiguous amino acid residues, at least 80 contiguous amino acid residues, at least 90 contiguous amino acid residues, at least contiguous 100 amino acid residues, at least 125 contiguous amino acid residues, at least 150 contiguous amino acid residues, at least 175 contiguous amino acid residues, at least 200 contiguous amino acid residues, or at least 250 contiguous amino acid residues of the amino acid sequence of a polypeptide or an antibody that immunospecifically binds to a polypeptide. In a specific embodiment, a fragment of a polypeptide or an antibody of that immunospecifically binds to an antigen retains at least 1, at least 2, or at least 3 functions of the polypeptide or antibody.

[0053] The term “fusion protein” as used herein refers to a polypeptide that comprises an amino acid sequence of an antibody and an amino acid sequence of a heterologous polypeptide or protein (i.e., a polypeptide or protein not normally a part of the antibody (e.g., a non-anti-RSV antigen antibody)).

[0054] The terms “high concentration” and “concentrated antibody” as used herein refer to a concentration of 50 mg/ml or higher, preferably 95 mg/ml or higher of an antibody or antigen-binding fragment thereof in an antibody formulation.

[0055] The term “high potency” as used herein refers to antibodies that exhibit high potency as determined in various assays for biological activity (e.g., neutralization of RSV) such as those described herein. For example, high potency antibodies of the invention have an IC₅₀ value less than 5 nM, less than 4 nM, less than 3 nM, less than 2 nM, less than 1.75 nM, less than 1.5 nM, less than 1.25 nM, less than 1 nM, less than 0.75 nM, less than 0.5 nM, less than 0.25 nM, less than 0.1 nM, less than 0.05 nM, less than 0.025 nM, or less than 0.01 nM, as measured by a microneutralization assay described herein. Further, high potency anti-RSV antibodies of the invention result in at least a 75%, preferably at least a 95% and more preferably a 99% lower RSV titer in a cotton rat 5 days after challenge with 10⁵ pfu relative to a cotton rat not administered said antibodies. In certain embodiments of the invention, high potency anti-RSV antibodies of the present invention exhibit a high affinity and/or high avidity for one or more RSV antigens (e.g., antibodies having an affinity of at least 2×10⁸ M⁻¹, preferably at least 2.5×10⁸ M⁻¹, at least 5×10⁸ M⁻¹, at least 10⁹ M⁻¹, at least 5×10⁹ M⁻¹, at least 10¹⁰ M⁻¹, at least 5×10¹⁰ M⁻¹, at least 10¹¹ M⁻¹, at least 5×10¹¹ M⁻¹, or at least 10¹² M⁻¹, or at least 5×10¹² M⁻¹ for one or more RSV antigens).

[0056] The term “host” as used herein refers to an animal, preferably a mammal, and most preferably a human.

[0057] The term “host cell” as used herein refers to the particular subject cell transfected with a nucleic acid molecule and the progeny or potential progeny of such a cell. Progeny of such a cell may not be identical to the parent cell transfected with the nucleic acid molecule due to mutations or environmental influences that may occur in succeeding generations or integration of the nucleic acid molecule into the host cell genome.

[0058] The term “human infant” as used herein refers to a human less than 24 months, preferably less than 16 months, less than 12 months, less than 6 months, less than 3 months, less than 2 months, or less than 1 month of age.

[0059] The term “human infant born prematurely” as used herein refers to a human born at less than 40 weeks gestational age, preferably less than 35 weeks gestational age, who is less than 6 months old, preferably less than 3 months old, more preferably less than 2 months old, and most preferably less than 1 month old.

[0060] As used herein, the term “in combination” refers to the use of more than one therapy. The use of the term “in combination” does not restrict the order in which therapies are administered to a subject with an infection. A first therapy can be administered before (e.g., 1 minute, 45 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks), concurrently, or after (e.g., 1 minute, 45 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks) the administration of a second therapy to a subject which had, has, or is susceptible to a disease or disorder. Any additional therapy can be administered in any order with the other additional therapies. In certain embodiments, the antibodies of the invention can be administered in combination with one or more non-antibody therapies. Non-limiting examples of therapies that can be administered in combination with an antibody of the invention include analgesic agents, anesthetic agents, antibiotics, or immunomodulatory agents.

[0061] As used herein, the term “infection” refers to all stages of RSV's life cycle in a host (including, but not limited to the invasion by and replication of RSV in a cell or body tissue), and the pathological state resulting from the invasion by and replication of a RSV. The invasion by and multiplication of a RSV includes, but is not limited to, the following steps: the docking of the RSV particle to a cell, the introduction of viral genetic information into a cell, the expression of RSV proteins, the production of new RSV particles and the release of RSV particles from a cell.

[0062] The term “inorganic salt” as used herein refers to any compounds containing no carbon that result from replacement of part or all of the acid hydrogen or an acid by a metal or a group acting like a metal and are often used as a tonicity adjusting compound in pharmaceutical compositions and preparations of biological materials. The most common inorganic salts are NaCl, KCl, NaH₂PO₄, etc.

[0063] An “isolated” or “purified” antibody is substantially free of cellular material or other contaminating proteins from the cell or tissue source from which the protein is derived, or substantially free of chemical precursors or other chemicals when chemically synthesized. The language “substantially free of cellular material” includes preparations of an antibody in which the antibody is separated from cellular components of the cells from which it is isolated or recombinantly produced. Thus, an antibody that is substantially free of cellular material includes preparations of antibody having less than about 30%, 20%, 10%, or 5% (by dry weight) of heterologous protein (also referred to herein as a “contaminating protein”). When the antibody is recombinantly produced, it is also preferably substantially free of culture medium, i.e., culture medium represents less than about 20%, 10%, or 5% of the volume of the protein preparation. When the antibody is produced by chemical synthesis, it is preferably substantially free of chemical precursors or other chemicals, i.e., it is separated from chemical precursors or other chemicals which are involved in the synthesis of the protein. Accordingly such preparations of the antibody have less than about 30%, 20%, 10%, 5% (by dry weight) of chemical precursors or compounds other than the antibody of interest. In a preferred embodiment, antibodies of the invention are isolated or purified.

[0064] An “isolated” nucleic acid molecule is one which is separated from other nucleic acid molecules which are present in the natural source of the nucleic acid molecule. Moreover, an “isolated” nucleic acid molecule, such as a cDNA molecule, can be substantially free of other cellular material, or culture medium when produced by recombinant techniques, or substantially free of chemical precursors or other chemicals when chemically synthesized. In a specific embodiment, a nucleic acid molecule(s) encoding an antibody of the invention is isolated or purified.

[0065] The phrase “low to undetectable levels of aggregation” as used herein refers to samples containing no more than 5%, no more than 4%, no more than 3%, no more than 2%, no more than 1% and most preferably no more than 0.5% aggregation by weight of protein as measured by high performance size exclusion chromatography (HPSEC) or by a multi-sizer.

[0066] The term “low to undetectable levels of fragmentation” as used herein refers to samples containing equal to or more than 95%, 98%, 99%, 99.5% or 99.9% of the total protein, for example, as determined by AUC or LC-MS.

[0067] The term “lower respiratory” tract refers to the major passages and structures of the lower respiratory tract including the windpipe (trachea) and the lungs, including the bronchi, bronchioles, and alveoli of the lungs.

[0068] As used herein, the term “low tolerance” refers to a state in which the patient suffers from side effects from a therapy so that the patient does not benefit from and/or will not continue therapy because of the adverse effects and/or the harm from side effects outweighs the benefit of the therapy.

[0069] As used herein, the terms “manage”, “managing” and “management” refer to the beneficial effects that a subject derives from a therapy (e.g., a prophylactic or therapeutic agent), which does not result in a cure of the infection. In certain embodiments, a subject is administered one or more therapies (e.g., prophylactic or therapeutic agents) to “manage” a infection, one or more symptoms thereof, or a respiratory condition associated with, potentiated by, or potentiating a RSV infection, so as to prevent the progression or worsening of the infection.

[0070] The terms “non-responsive” and “refractory” as used herein describe patients treated with a currently available therapy (such as but not limited to, a prophylactic or therapeutic agent) for a RSV infection, one or more symptoms thereof, or a respiratory condition associated with, potentiated by, or potentiating a RSV infection, which is not clinically adequate to relieve one or more symptoms associated with the infection. Typically, such patients suffer from severe, persistently active infection and require additional therapy to ameliorate the symptoms associated with their infection or respiratory condition.

[0071] As used herein, the terms “nucleic acids” and “nucleotide sequences” include DNA molecules (e.g., cDNA or genomic DNA), RNA molecules (e.g., mRNA), combinations of DNA and RNA molecules or hybrid DNA/RNA molecules, and analogues of DNA or RNA molecules. Such analogues can be generated using, for example, nucleotide analogues, which include, but are not limited to, inosine or tritylated bases. Such analogues can also comprise DNA or RNA molecules comprising modified backbones that lend beneficial attributes to the molecules such as, for example, nuclease resistance or an increased ability to cross cellular membranes. The nucleic acids or nucleotide sequences can be single-stranded, double-stranded, may contain both single-stranded and double-stranded portions, and may contain triple-stranded portions, but preferably is double-stranded DNA.

[0072] The term “pharmaceutically acceptable” as used herein means being approved by a regulatory agency of the Federal or a state government, or listed in the U.S. Pharmacopia, European Pharmacopia or other generally recognized pharmacopia for use in animals, and more particularly in humans.

[0073] The term “polyol” as used herein refers to a sugar that contains many —OH groups compared to a normal saccharide.

[0074] As used herein, the terms “prevent,” “preventing,” and “prevention” refer to the prevention or inhibition of the development or onset of a disease or disorder, such as an upper and/or lower respiratory tract RSV infection, otitis media or a respiratory condition related thereto in a subject, the prevention or inhibition of the progression of an upper respiratory tract RSV infection to a lower respiratory tract RSV infection, otitis media or a respiratory condition related thereto resulting from the administration of a therapy (e.g., a prophylactic or therapeutic agent), the prevention of a symptom of an upper and/or lower tract RSV infection, otitis media or a respiratory condition related thereto, or the administration of a combination of therapies (e.g., a combination of prophylactic or therapeutic agents).

[0075] As used herein, the term “prophylactic agent” refers to any agent that can prevent the recurrence, spread or onset of a disease or disorder, such as an upper and/or lower respiratory tract RSV infection, otitis media, or a symptom or respiratory condition relating thereto (including, but not limited to, asthma, wheezing, RAD, or a combination thereof), and/or prevent the progression of an upper respiratory tract RSV infection to a lower respiratory tract RSV infection or otitis media. In certain embodiments, the term “prophylactic agent” refers to an antibody of the invention. In certain other embodiments, the term “prophylactic agent” refers to an agent other than an antibody of the invention. Preferably, a prophylactic agent is an agent which is known to be useful to or has been or is currently being used to prevent or impede the onset, development, progression and/or severity of a RSV infection (preferably an upper and/or lower respiratory tract RSV infection) otitis media, and/or a symptom or respiratory condition related thereto.

[0076] In certain embodiments of the invention, a “prophylactically effective serum titer” is the serum titer in a subject, preferably a human, that reduces the incidence of an upper and/or lower respiratory tract RSV infection, otitis media and/or a symptom or respiratory condition related thereto in said subject. In some embodiments, the prophylactically effective serum titer prevents the progression of an upper respiratory tract RSV infection to a lower respiratory tract RSV infection or otitis media. Preferably, the prophylactically effective serum titer reduces the incidence of RSV infections in humans with the greatest probability of complications resulting from RSV infection (e.g., a human with cystic fibrosis, bronchopulmonary dysplasia, congenital heart disease, congenital immunodeficiency or acquired immunodeficiency, a human who has had a bone marrow transplant, a human infant, or an elderly human). In certain other embodiments of the invention, a “prophylactically effective serum titer” is the serum titer in a cotton rat that results in a RSV titer 5 days after challenge with 10⁵ pfu that is 99% lower than the RSV titer 5 days after challenge with 10⁵ pfu of RSV in a cotton rat not administered an antibody that immunospecifically binds to a RSV antigen.

[0077] As used herein, the term “refractory” refers to an upper and/or lower respiratory tract RSV infection, otitis media or a respiratory condition related thereto that is not responsive to one or more therapies (e.g., currently available therapies). In a certain embodiment, an upper and/or lower respiratory tract RSV infection, otitis media or a respiratory condition related thereto is refractory to a therapy means that at least some significant portion of the symptoms associated with said upper and/or lower respiratory tract RSV infection, otitis media or a respiratory condition related thereto are not eliminated or lessened by that therapy. The determination of whether an upper and/or lower respiratory tract RSV infection, otitis media or a respiratory condition related thereto is refractory can be made either in vivo or in vitro by any method known in the art for assaying the effectiveness of therapy for the infection, otitis media or the respiratory condition related thereto.

[0078] The term “RSV antigen” refers to a RSV polypeptide to which an antibody immunospecifically binds. A RSV antigen also refers to an analog or derivative of a RSV polypeptide or fragment thereof to which an antibody immunospecifically binds.

[0079] The term “serum titer” as used herein refers to an average serum titer in a population of least 10, preferably at least 20, and most preferably at least 40 subjects.

[0080] The term “saccharide” as used herein refers to a class of molecules that are derivatives of polyhydric alcohols. Saccharides are commonly referred to as carbohydrates and may contain different amounts of sugar (saccharide) units, e.g., monosaccharides, disaccharides and polysaccharides.

[0081] As used herein, the term “side effects” encompasses unwanted and adverse effects of a therapy (e.g., a prophylactic or therapeutic agent). Adverse effects are always unwanted, but unwanted effects are not necessarily adverse. An adverse effect from a therapy (e.g., a prophylactic or therapeutic agent) might be harmful or uncomfortable or risky. Examples of side effects include, but are not limited to, nausea, vomiting, anorexia, abdominal cramping, fever, pain, loss of body weight, dehydration, alopecia, dyspnea, insomnia, dizziness, mucositis, nerve and muscle effects, fatigue, dry mouth, and loss of appetite, rashes or swellings at the site of administration, flu-like symptoms such as fever, chills and fatigue, digestive tract problems and allergic reactions. Additional undesired effects experienced by patients are numerous and known in the art. Many are described in the Physician's Desk Reference (58^th ed., 2004).

[0082] The terms “stability” and “stable” as used herein in the context of a formulation comprising an antibody or antigen-binding fragment refer to the resistance of the antibody or antibody fragment in the formulation to thermal and chemical unfolding, aggregation, degradation or fragmentation under given manufacture, preparation, transportation and storage conditions. The “stable” formulations of the invention retain biological activity equal to or more than 80%, 85%, 90%, 95%, 98%, 99%, 99.5%, or 99.9% under given manufacture, preparation, transportation and storage conditions. The stability of the antibody or antibody fragment can be assessed by degrees of aggregation, degradation or fragmentation or levels of particular fragments (e.g., Fragment Type I or Fragment Type II) or types or sizes of aggregates by methods known to those skilled in the art, including but not limited to reduced AUC, SEC, LC-MS, particle multisizer Capillary Gel Electrophoresis (rCGE), Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) and HPSEC, compared to a reference, for example, a commercially available lyophilized palivizumab reconstituted to 100 mg/ml in 50 mM histidine/3.2 mM glycine buffer with 6% mannitol at pH 6.0. The reference regularly gives a single peak (≧97% area) by HPSEC. The overall stability of a formulation comprising an antibody or fragment thereof that immunospecifically binds to a RSV antigen can be assessed by various immunological assays including, for example, ELISA and radioimmunoassay, using the specific epitope of RSV.

[0083] As used herein, the terms “subject” and “patient” are used interchangeably. As used herein, a subject is preferably a mammal such as a non-primate (e.g., cows, pigs, horses, cats, dogs, rats, etc.) and a primate (e.g., monkey and human), most preferably a human. In one embodiment, the subject is a mammal, preferably a human, with an upper and/or lower respiratory tract RSV infection or otitis media. In another embodiment, the subject is a mammal, preferably a human, at risk of developing an upper and/or lower respiratory tract RSV infection or otitis media (e.g., an immunocompromised or immunosuppressed mammal, or a genetically predisposed mammal). In one embodiment, the subject is a human with a respiratory condition (including, but not limited to asthma, wheezing or RAD) that stems from, is caused by or associated with a RSV infection.

[0084] As used herein, the term “palivizumab standard reference” or analogous terms refer to commercially available lyophilized palivizumab, as described in the Physicians' Desk Reference, 56^th edition, 2002. Reconstituted palivizumab may contain, e.g., the following excipients: 47 mM histidine, 3.0 mM glycine and 5.6% manitol and the active ingredient, the antibody, at a concentration of 100 milligrams per ml solution.

[0085] As used herein, the terms “subject” and “patient” are used interchangeably. As used herein, the terms “subject” and “subjects” refer to an animal, preferably a mammal including a non-primate (e.g., a cow, pig, horse, cat, dog, rat, and mouse) and a non-primate (e.g., a monkey such as a cynomolgous monkey and a human), and more preferably a human.

[0086] The term “substantially free of surfactant” as used herein refers to a formulation of an antibody or fragment thereof that immunospecifically binds to a RSV antigen, said formulation containing less than 0.0005%, less than 0.0003%, or less than 0.0001% of surfactants and/or less than 0.0005%, less than 0.0003%, or less than 0.0001% of surfactants.

[0087] The term “substantially free of salt” as used herein refers to a formulation of an antibody or fragment thereof that immunospecifically binds to a RSV antigen, said formulation containing less than 0.0005%, less than 0.0003%, or less than 0.0001% of inorganic salts.

[0088] The term “surfactant” as used herein refers to organic substances having amphipathic structures; namely, they are composed of groups of opposing solubility tendencies, typically an oil-soluble hydrocarbon chain and a water-soluble ionic group. Surfactants can be classified, depending on the charge of the surface-active moiety, into anionic, cationic, and nonionic surfactants. Surfactants are often used as wetting, emulsifying, solubilizing, and dispersing agents for various pharmaceutical compositions and preparations of biological materials.

[0089] The term “synergistic” as used herein refers to a combination of therapies (e.g., use of prophylactic or therapeutic agents) which is more effective than the additive effects of any two or more single therapy. For example, a synergistic effect of a combination of prophylactic or therapeutic agents permits the use of lower dosages of one or more of the agents and/or less frequent administration of said agents to a subject with a RSV infection. The ability to utilize lower dosages of prophylactic or therapeutic therapies and/or to administer said therapies less frequently reduces the toxicity associated with the administration of said therapies to a subject without reducing the efficacy of said therapies in the prevention, management or treatment of a RSV infection. In addition, a synergistic effect can result in improved efficacy of therapies in the prevention or treatment of a RSV infection. Finally, synergistic effect of a combination of therapies (e.g., prophylactic or therapeutic agents) may avoid or reduce adverse or unwanted side effects associated with the use of any single therapy.

[0090] As used herein, the term “therapeutic agent” refers to any agent that can be used in the treatment, management, prevention or amelioration of a disease or disorder, for example, an upper and/or lower respiratory tract RSV infection, otitis media or a respiratory condition related thereto. In certain embodiments, the term “therapeutic agent” refers to an antibody of the invention. In certain other embodiments, the term “therapeutic agent” refers to an agent other than an antibody of the invention. Preferably, a therapeutic agent is an agent which is known to be useful for, or has been or is currently being used for the prevention, treatment, management or amelioration of a RSV infection (i.e., an upper and/or lower respiratory tract RSV infection), otitis media, or one or more symptoms or respiratory conditions related thereto.

[0091] In certain embodiments of the invention, a “therapeutically effective serum titer” is the serum titer in a subject, preferably a human, that reduces the severity, the duration and/or the symptoms associated with a RSV infection in said subject. Preferably, the therapeutically effective serum titer reduces the severity, the duration and/or the number symptoms associated with upper and/or lower respiratory tract RSV infections in humans with the greatest probability of complications resulting from the infection (e.g., a human with cystic fibrosis, bronchopulmonary dysplasia, congenital heart disease, congenital immunodeficiency or acquired immunodeficiency, a human who has had a bone marrow transplant, a human infant, or an elderly human). In certain other embodiments of the invention, a “therapeutically effective serum titer” is the serum titer in a cotton rat that results in a RSV titer 5 days after challenge with 10⁵ pfu that is 99% lower than the RSV titer 5 days after challenge with 10⁵ pfu of RSV in a cotton rat not administered an antibody that immunospecifically binds to a RSV antigen.

[0092] As used herein, the term “therapy” refers to any protocol, method and/or agent that can be used in the prevention, treatment or management of a disease or disorder, such as an RSV infection (i.e., an upper and/or lower respiratory tract RSV infection), otitis media, or a symptom or respiratory condition relating thereto (including, but not limited to, asthma, wheezing, RAD, or a combination thereof). In certain embodiments, the terms “therapies” and “therapy” refer to a biological therapy, supportive therapy, and/or other therapies useful in the treatment, management, prevention and/or amelioration of a RSV infection (i.e., an upper and/or lower respiratory tract RSV infection), otitis media, or a symptom or respiratory condition relating thereto (including, but not limited to, asthma, wheezing, RAD, or a combination thereof) known to one of skill in the art such as medical personnel.

[0093] As used herein, the terms “treat,” “treatment” and “treating” refer to the reduction or amelioration of the progression, severity, and/or duration of a disease or disorder, such as an upper and/or lower respiratory tract RSV infection, otitis media, or a symptom or respiratory condition related thereto (such as asthma, wheezing, RAD, or a combination thereof) resulting from the administration of one or more therapies (including, but not limited to, the administration of one or more prophylactic or therapeutic agents). In specific embodiments, such terms refer to the reduction or inhibition of the replication of RSV, the inhibition or reduction in the spread of RSV to other tissues or subjects (e.g., the spread to the lower respiratory tract), the inhibition or reduction of infection of a cell with a RSV, or the amelioration of one or more symptoms associated with an upper and/or lower respiratory tract RSV infection or otitis media.

[0094] The term “upper and/or lower respiratory” tract refers to the major passages and structures of the upper and/or lower respiratory tract including the nose or nostrils, nasal cavity, mouth, throat (pharynx), and voice box (larynx).

[0095] FIG. 1 is a schematic diagram showing an outline for preparing purified antibodies that immunospecifically bind to RSV antigen.

[0096] FIG. 2 is a schematic diagram showing an outline for preparing purified antibodies that immunospecifically bind to RSV antigen.

[0097] FIG. 3A-3B show the amino acid sequences of the (A) light chain variable region and (B) heavy chain variable region of a monoclonal antibody that binds to a RSV antigen, the potency of which can be increased by methods described herein or in Applicants' copending application Ser. Nos. 60/168,426 and 60/186,252 and U.S. Pat. No. 6,656,467. For reference purposes, this is the amino acid sequence of the palivizumab antibody disclosed in Johnson et al., 1997, J. Infect. Dis. 176:1215-1224 and U.S. Pat. No. 5,824,307. Here, the CDR regions are underlined while non-underlined residues form the framework (FR) regions of the variable regions of the antibody. In this antibody, the CDRs are derived from a mouse antibody while the framework regions are derived from a human antibody. The constant regions (not shown) are also derived from a human antibody.

[0098] FIG. 4A-4B show the (A) light chain variable region and (B) heavy light chain variable region for an antibody sequence. CDR regions are underlined, and the non-underlined residues form the framework of the variable regions of the antibody. This sequence differs from the sequence disclosed in FIGS. 1A-1B in the first 4 residues of VH CDR1 of the light chain, residue 103 of the light chain FR4 and residue 112 of the heavy chain FR4. For reference purposes, these VL and VH sequences are identical to the VL and VH domains of IX-493L1FR (see Table 2).

[0099] FIG. 5A-5B show the nucleotide and translated amino acid sequence of the A4B4L1FR-S28R (A) VH domain and (B) VL domain. CDR sequences are underlined. Where palivizumab differs from A4B4L1FR-S28R, the palivizumab amino acid is shown below the motavizumab sequence. Residues that were introduced on the IX-493L1FR template (see also FIG. 2) are indicated in bold.

[0100] FIG. 6A-6C. Quantitation of aggregates, fragments and monomers of A4B4L1FR-S28R during storage at (♦) 2-8° C., (□) 20-24° C. and (▴) 38-42° C.; as determined by SEC with UV detection. (A) Percent Aggregates; (B) Percent Fragments and (C) Percent Purity (monomers).

[0101] FIG. 7. Plot of Aggregation and fragmentation rates of A4B4L1FR-S28R based on the SEC data of FIGS. 19A-19C; (♦) rate of aggregation, (▪) rate of fragmentation.

[0102] FIG. 8. SEC profile of A4B4L1FR-S28R formulated in 25 mM histidine-HCl, pH 6.0 after storage at 38-42° C. for one month.

[0103] FIG. 9. Comparison of AUC and SEC analysis of A4B4L1FR-S28R at initial, 9-month and 14-month time points. All samples were formulated in 25 mM histidine-HCl, pH 6.0 and, for the 9 and 14 moth points, stored at 38-42° C. (A) AUC; (B) SEC.

[0104] FIG. 10. Comparison of antibody sample concentration dependence of signal/noise ratio for AUC analysis.

[0105] FIG. 11. AUC analysis of A4B4L1FR-S28R formulated in 25 mM histidine-HCl, pH 6.0 and stored at 38-42° C. over the course of 5 days.

[0106] FIG. 12. LC-MS analysis of deglycosylated, reduced and alkylated antibody type I fragment. Sample collected from SEC of A4B4L1FR-S28R formulated in 25 mM histidine-HCl, pH 6.0 and stored at 38-42° C. for 1 month.

[0107] FIG. 13. LC-MS analysis of deglycosylated, reduced and alkylated antibody type II fragment. Sample collected from SEC of A4B4L1FR-S28R formulated in 25 mM histidine-HCl, pH 6.0 and stored at 38-42° C. for 1 month.

[0108] FIG. 14A-14B is a diagram showing the characteristic fragmentation pattern of A4B4L1FR-S28R, forming antibody type I and antibody type II fragments. (A) Cleavage sites within the hinge region of the antibody heavy chain. Bold arrows indicate preferred or predominant cleavage sites. (B) Schematic showing characteristics of antibody type I and antibody type II fragments. an outline for preparing purified antibodies that immunospecifically bind to RSV antigen. The antibody type I fragment comprises a full length antibody light chain, a full length antibody heavy chain and a C-terminal portion of an antibody heavy chain that, in human IgG₁ immunoglobulins, has an N-terminus at cysteine 223, aspartic acid 224, lysine 225, threonine 226, histidine 227, threonine 228 or cysteine 229. The antibody type II fragment comprises an antibody light chain and an N-terminal portion of an antibody heavy chain that, in human IgG₁ immunoglobulins, has a C-terminus at serine 222, cysteine 223, aspartic acid 224, lysine 225, threonine 226, histidine 227 or threonine 228.

[0109] FIG. 15. Chromatograms of Lys-C digested aggregates, monomers and fragments collected from SEC of A4B4L1FR-S28R formulated in 25 mM histidine-HCl, pH 6.0 and stored at 38-42° C. for 1 month. The arrows point to the low level disulfide bond scrambling peaks.

[0110] FIG. 16. Chromatograms of Lys-C digested aggregates with and without reduction. Samples were collected from SEC of A4B4L1FR-S28R formulated in 25 mM histidine-HCl, pH 6.0 and stored at 38-42° C. for 1 month. The arrows point to the low level disulfide bond scrambling peaks.

[0111] FIG. 17 summarizes the results of a RSV microneutralization assay using the anti-RSV antibodies A4B4L1FR-S28R and palivizumab, comparing the ability of both antibodies to inhibit the in vitro replication of RSV (Long) in the assay.

[0112] FIG. 18 summarizes the results of a RSV microneutralization assay demonstrating the ability of A4B4L1FR-S28R to inhibit the in vitro replication of RSV (Long) in the microneutralization assay.

[0113] FIG. 19 DSC thermograms of the full length palivizumab (top panel) and an overlay of the thermograms obtained from purified Fab and Fc fragments of palivizumab (bottom panel). Two discrete peaks are seen for the Fc domain at approximately 68° C. and 83° C. A single peak is seen for the Fab fragment at approximately 87° C.

[0114] FIG. 20 plot of the Tm and pI values of palivizumab and motavizumab.

[0115] FIG. 21 plot of the viscosity of a 100 mg/ml solution of palivizumab and motavizumab at a range of temperatures from about 2 to 25° C.

[0116] FIG. 22 plot of the aggregation rates of palivizumab and motavizumab against the Fab Tm for each antibody.

[0117] The present invention provides methods for preparing formulations of antibodies, or derivatives, analogues, or fragments thereof that immunospecifically bind to a an antigen of interest. Such antibodies may be purified according to any method known in the art for purification of antibodies. FIGS. 1 and 2 are schematic diagrams showing alternate outlines for preparing purified antibodies. In one embodiment, the methods for preparing liquid formulations of the present invention comprise: concentrating a fraction containing the purified antibody or a fragment to a final antibody or fragment concentration of from about 15 mg/ml, about 20 mg/ml, about 30 mg/ml, about 40 mg/ml, about 50 mg/ml, about 60 mg/ml, about 70 mg/ml, about 80 mg/ml, about 90 mg/ml, about 100 mg/ml, about 110 mg/ml, about 125 mg/ml, about 150 mg/ml, about 200 mg/ml, about 250 mg/ml, or about 300 mg/ml using a semipermeable membrane with an appropriate molecular weight (MW) cutoff (e.g., 30 kD cutoff for whole antibody molecules and F(ab′)₂ fragments; and 10 kD cutoff for antibody fragments, such as Fab fragments) and difiltrating the concentrated antibody fraction into the formulation buffer using the same membrane. Antibodies are preferably expressed in myeloma cells, more preferably murine myeloma cells, most preferably NSO cells.

[0118] In the embodiment outlined by FIG. 1, conditioned medium containing antibody or a fragment thereof that immunospecifically binds to an antigen of interest is subjected to CUNO filtration and the filtered antibody is subjected to HS50 cation exchange chromatography. The fraction from the HS50 cation exchange chromatography is then subjected to rProtein A affinity chromatography followed by low pH treatment. Following low pH treatment, the antibody fraction is subject to super Q 650 anion exchange chromatography and then nanofiltration. The fraction of the antibody obtained after nanofiltration is then subjected to diafiltration to concentrate the antibody fraction into the formulation buffer using the same membrane.

[0119] Using the embodiment of FIG. 2, conditioned medium containing antibody or a fragment thereof that immunospecifically binds to an antigen of interest is subjected to CUNO filtration and the filtered antibody is subjected to Fractogel® S cation exchange chromatography. The fraction from the cation exchange chromatography is then subjected to super Q anion chromatography. followed by nanofiltration with a Planova® 20 N nanofilter. The antibody fraction recovered after nanofiltration is then subjected to low pH treatment followed by hydroxyapatite (HA) chromatography. The fraction of the antibody obtained after HA chromatography is then subjected to diafiltration to concentrate the antibody fraction into the formulation buffer using the same membrane.

[0120] The formulation buffer of the present invention preferably comprises histidine at a concentration ranging from about 1 mM to about 100 mM, about 10 mM to about 50 mM, about 20 mM to about 30 mM, or about 23 mM to about 27 mM. Preferably, the formulation buffer of the present invention comprises histidine at a concentration of about 25 mM. The formulations may further comprise glycine at a concentration of less than 100 mM, less than 50 mM, less than 3.0 mM, less than 2.0 mM, or less than 1.8 mM. Preferably, the formulations comprise glycine at a concentration of 1.6 mM. The amount of glycine in the formulation should not cause a significant buffering in order to avoid antibody precipitation at its isoelectric point. The pH of the formulation may range from about 5.0 to about 7.0, preferably about 5.5 to about 6.5, more preferably about 5.8 to about 6.2, and most preferably about 6.0. To obtain an appropriate pH for a particular antibody, it is preferable that histidine (and glycine, if added) is first dissolved in water to obtain a buffer solution with higher pH than the desired pH and then the pH is brought down to the desired level by adding HCl. This way, the formation of inorganic salts (e.g., formation of NaCl when, for example, histidine hydrochloride is used as histidine and pH is raised to a desired level by adding NaOH) can be avoided.

[0121] The formulations of the present invention can be prepared as unit dosage forms by preparing a vial containing an aliquot of the liquid formulation for a one-time use. For example, a unit dosage per vial may contain 1 ml, 2 ml, 3 ml, 4 ml, 5 ml, 6 ml, 7 ml, 8 ml, 9 ml, 10 ml, 15 ml, or 20 ml of different concentrations of an antibody or a fragment thereof that immunospecifically binds to the antigen of interest ranging from about 15 mg/ml to about 300 mg/ml. If necessary, these preparations can be adjusted to a desired concentration by adding a sterile diluent to each vial.

[0122] The formulations of the present invention may be sterilized by various sterilization methods, including sterile filtration, radiation, etc. In a most preferred embodiment, the difiltrated antibody formulation is filter-sterilized with a presterilized 0.22-micron filter. In specific embodiments, sterilized liquid formulations of the present invention may be administered to a subject to prevent, treat, manage or ameliorate a RSV infection, one or more symptoms thereof, or a respiratory condition associated with, potentiated by, potentiating a RSV infection.

[0123] The formulations of the invention comprise labeled antibodies, derivatives and analogues thereof, that immunospecifically bind to an antigen of interest and can be used for diagnostic purposes to detect, diagnose, or monitor a disorder associated with and/or characterized by the presence or said antigen. In a specific embodiment, the formulations of the invention comprise labeled antibodies, derivatives and analogues thereof, that immunospecifically bind to a RSV antigen and can be used for diagnostic purposes to detect, diagnose, or monitor a RSV infection.

[0124] The invention encompasses both liquid and lyophilized forms of the formulations. Methods to produce lyophilized forms of liquid formulations are well-characterized in the art. In one embodiment, the ingredients of formulation of the invention are supplied either separately or mixed together in unit dosage form, for example, as a dry lyophilized powder or water free concentrate in a hermetically sealed container such as an ampoule or sachette indicating the quantity of active agent. Where the composition is to be administered by infusion, it can be dispensed with an infusion bottle containing sterile pharmaceutical grade water or saline. Where the composition is administered by injection, an ampoule of sterile water for injection or saline can be provided so that the ingredients may be mixed prior to administration.

[0125] The compositions of the invention can be formulated as neutral or salt forms. Pharmaceutically acceptable salts include those formed with anions such as those derived from hydrochloric, phosphoric, acetic, oxalic, tartaric acids, etc., and those formed with cations such as those derived from sodium, potassium, ammonium, calcium, ferric hydroxides, isopropylamine, triethylamine, 2-ethylamino ethanol, histidine, procaine, etc.

[0126] The formulation of the invention can be further processed into an oral or non-oral dosage form, for immediate or extended release. The formulation can additionally comprise inactive ingredients ordinarily used in pharmaceutical preparation such as diluents, fillers, disintegrants, sweeteners, lubricants and flavors. The formulation may also be processed for intravenous administration, either by bolus injection or sustained drip, or for release from an implanted capsule. A typical formulation for intravenous administration utilizes physiological saline as a diluent.

[0127] The invention provides formulations comprising antibodies of the invention for use in diagnosing, detecting, or monitoring a disorder, in preventing, treating, managing, or ameliorating of a disorder or one or more symptoms thereof, and/or in research. In a specific embodiment, the formulation of the invention comprises one or more antibodies. In another embodiment, the formulation of the invention comprises one or more antibodies and one or more prophylactic or therapeutic agents other than antibodies. Preferably, the prophylactic or therapeutic agents known to be useful for or having been or currently being used in the prevention, treatment, management, or amelioration of a disorder or one or more symptoms thereof. In accordance with these embodiments, the composition may further comprise of a carrier, diluent or excipient.

[0128] The formulations of the present invention provide antibody formulations which are substantially free of surfactant, inorganic salts, and/or other excipients and yet exhibit high stability during long periods of storage. In a specific embodiment, such antibody formulations are homogeneous. The formulations of the present invention comprise histidine at concentrations between 1 and 100 mM and an antibody which immunospecifically binds to a antigen of interest at concentrations of about 15 mg/ml to about 300 mg/ml. In one embodiment, the formulations of the invention do not comprise other ingredients except for water or suitable solvents. In a specific embodiment, the antibody immunospecifically binds to an RSV antigen and in preferred embodiments is not palivizumab or a fragment thereof.

[0129] In one embodiment, the antibody of the formulation of the invention is an antibody or antibody fragment conjugated to another moiety, including, but not limited to, a heterologous polypeptide, another antibody or another fragment, a marker sequence, a diagnostic agent, a therapeutic agent, a radioactive metal ion, a polymer, albumin, and a solid support. In another embodiment, formulations of the invention comprise two or more antibodies, or fragments thereof that immunospecifically binds to an antigen of interest. In a specific embodiment, formulations of the invention comprise two or more antibodies, or fragments thereof, that immunospecifically binds to a RSV antigen, wherein at least one of the antibodies or antibody fragments is not palivizumab or a fragment thereof.

[0130] The concentration of an antibody or a fragment thereof which is included in the formulations of the invention is at least 15 mg/ml, at least 20 mg/ml, at least 25 mg/ml, at least 30 mg/ml, at least 35 mg/ml, at least 40 mg/ml, at least 45 mg/ml, at least 50 mg/ml, at least 55 mg/ml, at least 60 mg/ml, at least 65 mg/ml, at least 70 mg/ml, at least 75 mg/ml, at least 80 mg/ml, at least 85 mg/ml, at least 90 mg/ml, at least 95 mg/ml, at least 100 mg/ml, at least 105 mg/ml, at least 110 mg/ml, at least 115 mg/ml, at least 120 mg/ml, at least 125 mg/ml, at least 130 mg/ml, at least 135 mg/ml, at least 140 mg/ml, at least 150 mg/ml, at least 200 mg/ml, at least 250 mg/ml, or at least 300 mg/ml.

[0131] The concentration of histidine which is included in the formulations of the invention ranges from about 1 mM to about 100 mM, about 10 mM to about 50 mM, about 20 mM to about 30 mM, or about 23 mM to about 27 mM, and is most preferably about 25 mM. Histidine can be in the form of L-histidine, D-histidine, or a mixture thereof, but L-histidine is the most preferable. Histidine can also be in the form of hydrates. Histidine may be used in a form of pharmaceutically acceptable salt, such as hydrochloride (e.g., monohydrochloride and dihydrochloride), hydrobromide, sulfate, acetate, etc. The purity of histidine should be at least 98%, preferably at least 99%, and most preferably at least 99.5%.

[0132] The pH of the formulation should not be equal to the isoelectric point of the particular antibody to be used in the formulation and may range from about 5.0 to about 7, preferably about 5.5 to about 6.5, more preferably about 5.8 to about 6.2, and most preferably about 6.0.

[0133] In addition to histidine and an antibody or a fragment thereof, the formulations of the present invention may further comprise glycine at a concentration of less than 100 mM, less than 50 mM, less than 3.0 mM, less than 2.0 mM, or less than 1.8 mM, and most preferably 1.6 mM. The amount of glycine in the formulation should not cause a significant buffering effect so that antibody precipitation at its isoelectric point can be avoided. Glycine may be also used in a form of pharmaceutically acceptable salt, such as hydrochloride, hydrobromide, sulfate, acetate, etc. The purity of glycine should be at least 98%, preferably at least 99%, and most preferably 99.5%. In a specific embodiment, glycine is included in the formulations of the present invention.

[0134] Optionally, the formulations of the present invention may further comprise other excipients, such as saccharides (e.g., sucrose, mannose, trehalose, etc.) and polyols (e.g., mannitol, sorbitol, etc.). In one embodiment, the other excipient is a saccharide. In a specific embodiment, the saccharide is sucrose, which is at a concentration ranging from between about 1% to about 20%, preferably about 5% to about 15%, and more preferably about 8% to 10%. In another embodiment, the other excipient is a polyol. Preferably, however, the liquid formulations of the present invention do not contain mannitol. In a specific embodiment, the polyol is polysorbate (e.g., Tween 20), which is at a concentration ranging from between about 0.001% to about 1%, preferably, about 0.01 to about 0.1.

[0135] The formulations of the present invention exhibit stability at the temperature ranges of 38° C.-42° C. for at least 60 days and, in some embodiments, at least 120 days, of 20° C.-24° C. for at least 1 year, of 2° C.-8° C. (in particular, at 4° C.) for at least 3 years, at least 4 years, or at least 5 years and at −20° C. for at least 3 years, at least 4 years, or at least 5 years, as assessed by AUC, LC-MS, size exclusion chromatography (SEC) or high performance size exclusion chromatography (HPSEC) or particle multisizer. Namely, the formulations of the present invention have low to undetectable levels of aggregation and/or fragmentation, as defined herein, after the storage for the defined periods as set forth above. Preferably, no more than 5%, no more than 4%, no more than 3%, no more than 2%, no more than 1%, and most preferably no more than 0.5% (but in certain embodiments, at least 0.1%) of the antibody or antibody fragment forms an aggregate or fragment (particularly of fragment I or fragment II) as measured by AUC, LC-MS, SEC or HPSEC, after the storage for the defined periods as set forth above. Furthermore, formulations of the present invention exhibit almost no loss in biological activities of the antibody or antibody fragment during the prolonged storage under the condition described above, as assessed by various immunological assays including, but not limited to, enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay to measure the ability of the antibody or antibody fragment to immunospecifically bind to an antigen of interest, and by a C3a/C4a assay to measure the complement activating ability of the antibody. The formulations of the present invention retain after the storage for the above-defined periods more than 80%, more than 85%, more than 90%, more than 95%, more than 98%, more than 99%, or more than 99.5% of the initial biological activities of the formulation prior to the storage.

[0136] The formulations of the present invention can be prepared as unit dosage forms. For example, a unit dosage per vial may contain 1 ml, 2 ml, 3 ml, 4 ml, 5 ml, 6 ml, 7 ml, 8 ml, 9 ml, 10 ml, 15 ml, or 20 ml of different concentrations of an antibody or a fragment thereof that immunospecifically binds to a RSV antigen ranging from about 15 mg/ml to about 300 mg/ml. If necessary, these preparations can be adjusted to a desired concentration by adding a sterile diluent to each vial.

[0137] The invention encompasses stable liquid formulations comprising a single antibody or fragment thereof that immunospecifically binds to an antigen of interest. In a specific embodiment, the invention encompasses stable liquid formulations comprising a single antibody or fragment thereof that immunospecifically binds to a RSV antigen, with the proviso that said antibody is not palivizumab. The invention also encompasses stable liquid formulations comprising two or more antibodies or fragments thereof that immunospecifically bind to a RSV antigen. In one embodiment, a stable liquid formulation of the invention comprises two or more antibodies or fragments thereof that immunospecifically bind to a RSV antigen, wherein one of the antibodies or antibody fragments is not palivizumab or a fragment thereof.

[0138] The antibodies useful in the present invention include, but are not limited to, monoclonal antibodies, synthetic antibodies, multispecific antibodies (including bi-specific antibodies), human antibodies, humanized antibodies, chimeric antibodies, single-chain Fvs (scFv) (including bi-specific scFvs), single chain antibodies, Fab fragments, F(ab′) fragments, disulfide-linked Fvs (sdFv), and epitope-binding fragments of any of the above. In particular, antibodies of the present invention include immunoglobulin molecules and immunologically active portions of immunoglobulin molecules, i.e., molecules that contain an antigen binding site that immunospecifically binds to an antigen. The immunoglobulin molecules of the invention can be of any type (e.g., IgG, IgE, IgM, IgD, IgA and IgY), class (e.g., IgG₁, IgG₂, IgG₃, IgG₄, IgA₁, and IgA₂) or subclass of immunoglobulin molecule. Preferably, the antibodies of the invention are IgG, more preferably, IgG₁.

[0139] The antibodies useful in the present invention may be from any animal origin including birds and mammals (e.g., human, murine, donkey, sheep, rabbit, goat, guinea pig, camel, horse, or chicken). Preferably, the antibodies are human or humanized monoclonal antibodies. As used herein, “human” antibodies include antibodies having the amino acid sequence of a human immunoglobulin and include antibodies isolated from human immunoglobulin libraries or from mice or other animal that express antibodies from human genes.

[0140] The antibodies useful in the present invention may be monospecific, bispecific, trispecific or of greater multispecificity. Multispecific antibodies may immunospecifically bind to different epitopes of a polypeptide or may immunospecifically bind to both a polypeptide as well a heterologous epitope, such as a heterologous polypeptide or solid support material. See, e.g., International Publication Nos. WO 93/17715, WO 92/08802, WO 91/00360, and WO 92/05793; Tutt, et al., 1991, J. Immunol. 147:60-69; U.S. Pat. Nos. 4,474,893, 4,714,681, 4,925,648, 5,573,920, and 5,601,819; and Kostelny et al., 1992, J. Immunol. 148:1547-1553.

[0141] The antibodies useful in the present invention include derivatives of the antibodies. Standard techniques known to those of skill in the art can be used to introduce mutations in the nucleotide sequence encoding an antibody to be used with the methods of the invention, including, for example, site-directed mutagenesis and PCR-mediated mutagenesis which result in amino acid substitutions. Preferably, the derivatives include less than 25 amino acid substitutions, less than 20 amino acid substitutions, less than 15 amino acid substitutions, less than 10 amino acid substitutions, less than 5 amino acid substitutions, less than 4 amino acid substitutions, less than 3 amino acid substitutions, or less than 2 amino acid substitutions relative to the original molecule. In a preferred embodiment, the derivatives have conservative amino acid substitutions are made at one or more predicted non-essential amino acid residues. A “conservative amino acid substitution” is one in which the amino acid residue is replaced with an amino acid residue having a side chain with a similar charge. Families of amino acid residues having side chains with similar charges have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Alternatively, mutations can be introduced randomly along all or part of the coding sequence, such as by saturation mutagenesis, and the resultant mutants can be screened for biological activity to identify mutants that retain activity. Following mutagenesis, the encoded protein can be expressed and the activity of the protein can be determined.

[0142] The antibodies useful in the present invention include derivatives that are modified, i.e, by the covalent attachment of any type of molecule to the antibody such that covalent attachment. For example, but not by way of limitation, the antibody derivatives include antibodies that have been modified, e.g., by glycosylation, acetylation, pegylation, phosphorylation, amidation, derivatization by known protecting/blocking groups, proteolytic cleavage, linkage to a cellular ligand or other protein, etc. Any of numerous chemical modifications may be carried out by known techniques, including, but not limited to specific chemical cleavage, acetylation, formylation, synthesis in the presence of tunicamycin, etc. Additionally, the derivative may contain one or more non-classical amino acids.

[0143] Antibodies useful in the present invention or fragments thereof can also comprise a framework region known to those of skill in the art. In certain embodiments, one or more framework regions, preferably, all of the framework regions, of an antibody to be used in the methods of the invention or fragment thereof are human. In certain other embodiments of the invention, the fragment region of an antibody of the invention or fragment thereof is humanized. In certain embodiments, the antibody to be used with the methods of the invention is a synthetic antibody, a monoclonal antibody, an intrabody, a chimeric antibody, a human antibody, a humanized chimeric antibody, a humanized antibody, a glycosylated antibody, a multispecific antibody, a human antibody, a single-chain antibody, or a bispecific antibody.

[0144] In certain embodiments of the invention, the antibodies useful in the present invention have half-lives in a mammal, preferably a human, of greater than 12 hours, greater than 1 day, greater than 3 days, greater than 6 days, greater than 10 days, greater than 15 days, greater than 20 days, greater than 25 days, greater than 30 days, greater than 35 days, greater than 40 days, greater than 45 days, greater than 2 months, greater than 3 months, greater than 4 months, or greater than 5 months. Antibodies or antigen-binding fragments thereof having increased in vivo half-lives can be generated by techniques known to those of skill in the art. For example, antibodies or antigen-binding fragments thereof with increased in vivo half-lives can be generated by modifying (e.g., substituting, deleting or adding) amino acid residues identified as involved in the interaction between the Fc domain and the FcRn receptor (see, e.g., PCT Publication No. WO 97/34631 and U.S. patent application Ser. No. 10/020,354, entitled “Molecules with Extended Half-Lives, Compositions and Uses Thereof”, filed Dec. 12, 2001, by Johnson et al., which are incorporated herein by reference in their entireties). Such antibodies or antigen-binding fragments thereof can be tested for binding activity to RSV antigens as well as for in vivo efficacy using methods known to those skilled in the art, for example, by immunoassays described herein.

[0145] Further, antibodies or antigen-binding fragments thereof with increased in vivo half-lives can be generated by attaching to said antibodies or antibody fragments polymer molecules such as high molecular weight polyethyleneglycol (PEG). PEG can be attached to said antibodies or antibody fragments with or without a multifunctional linker either through site-specific conjugation of the PEG to the N- or C-terminus of said antibodies or antibody fragments or via epsilon-amino groups present on lysine residues. Linear or branched polymer derivatization that results in minimal loss of biological activity will be used. The degree of conjugation will be closely monitored by SDS-PAGE and mass spectrometry to ensure proper conjugation of PEG molecules to the antibodies. Unreacted PEG can be separated from antibody-PEG conjugates by, e.g., size exclusion or ion-exchange chromatography. PEG-derivatizated antibodies or antigen-binding fragments thereof can be tested for binding activity to RSV antigens as well as for in vivo efficacy using methods known to those skilled in the art, for example, by immunoassays described herein.

[0146] The antibodies useful in the present invention can be single-chain antibodies. The design and construction of a single-chain antibody is described in Marasco et al, 1993, Proc Natl Acad Sci 90:7889-7893, which is incorporated herein by reference in its entirety.

[0147] In certain embodiments, the antibodies useful in the present invention bind to an intracellular epitope, i.e., are intrabodies. An intrabody comprises at least a portion of an antibody that is capable of immunospecifically binding an antigen and preferably does not contain sequences coding for its secretion. Such antibodies will bind its antigen intracellularly. In one embodiment, the intrabody comprises a single-chain Fv (“sFv”). sFv are antibody fragments comprising the V_H and V_L domains of antibody, wherein these domains are present in a single polypeptide chain. Generally, the Fv polypeptide further comprises a polypeptide linker between the V_H and V_L domains which enables the sFv to form the desired structure for antigen binding. For a review of sFv see Pluckthun in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds. Springer-Verlag, New York, pp. 269-315 (1994).

[0148] In a further embodiment, the intrabody preferably does not encode an operable secretory sequence and thus remains within the cell (see generally Marasco, W A, 1998, “Intrabodies: Basic Research and Clinical Gene Therapy Applications” Springer:New York).

[0149] The present invention also encompasses formulations comprising antibodies that are conjugated or fused to one or more moieties, including but not limited to, peptides, polypeptides, proteins, fusion proteins, nucleic acid molecules, small molecules, mimetic agents, synthetic drugs, inorganic molecules, and organic molecules.

[0150] The present invention encompasses formulations comprising antibodies that are recombinantly fused or chemically conjugated (including both covalent and non-covalent conjugations) to a heterologous protein or polypeptide (or fragment thereof, preferably to a polypeptide of at least 10, at least 20, at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, at least 90 or at least 100 amino acids) to generate fusion proteins. The fusion does not necessarily need to be direct, but may occur through linker sequences. For example, antibodies may be used to target heterologous polypeptides to particular cell types, either in vitro or in vivo, by fusing or conjugating the antibodies to antibodies specific for particular cell surface receptors. Antibodies fused or conjugated to heterologous polypeptides may also be used in in vitro immunoassays and purification methods using methods known in the art. See e.g., International publication No. WO 93/21232; European Patent No. EP 439,095; Naramura et al., 1994, Immunol. Lett. 39:91-99; U.S. Pat. No. 5,474,981; Gillies et al., 1992, PNAS 89:1428-1432; and Fell et al., 1991, J. Immunol. 146:2446-2452, which are incorporated by reference in their entireties.

[0151] The present invention further includes formulations comprising heterologous proteins, peptides or polypeptides fused or conjugated to antibody fragments. For example, the heterologous polypeptides may be fused or conjugated to a Fab fragment, Fd fragment, Fv fragment, F(ab)₂ fragment, a VH domain, a VL domain, a VH CDR, a VL CDR, or fragment thereof. Methods for fusing or conjugating polypeptides to antibody portions are well-known in the art. See, e.g., U.S. Pat. Nos. 5,336,603, 5,622,929, 5,359,046, 5,349,053, 5,447,851, and 5,112,946; European Patent Nos. EP 307,434 and EP 367,166; International publication Nos. WO 96/04388 and WO 91/06570; Ashkenazi et al., 1991, Proc. Natl. Acad. Sci. USA 88: 10535-10539; Zheng et al., 1995, J. Immunol. 154:5590-5600; and Vil et al., 1992, Proc. Natl. Acad. Sci. USA 89:11337-11341 (said references incorporated by reference in their entireties).

[0152] Additional fusion proteins may be generated through the techniques of gene-shuffling, motif-shuffling, exon-shuffling, and/or codon-shuffling (collectively referred to as “DNA shuffling”). DNA shuffling may be employed to alter the activities of antibodies of the invention or fragments thereof (e.g., antibodies or fragments thereof with higher affinities and lower dissociation rates). See, generally, U.S. Pat. Nos. 5,605,793; 5,811,238; 5,830,721; 5,834,252; and 5,837,458, and Patten et al., 1997, Curr. Opinion Biotechnol. 8:724-33; Harayama, 1998, Trends Biotechnol. 16(2):76-82; Hansson, et al., 1999, J. Mol. Biol. 287:265-76; and Lorenzo and Blasco, 1998, Biotechniques 24(2):308-313 (each of these patents and publications are hereby incorporated by reference in its entirety). Antibodies or fragments thereof, or the encoded antibodies or fragments thereof, may be altered by being subjected to random mutagenesis by error-prone PCR, random nucleotide insertion or other methods prior to recombination. One or more portions of a polynucleotide encoding an antibody or antibody fragment may be recombined with one or more components, motifs, sections, parts, domains, fragments, etc. of one or more heterologous molecules.

[0153] Moreover, the antibodies or fragments thereof can be fused to marker sequences, such as a peptide to facilitate purification. In embodiments, the marker amino acid sequence is a hexa-histidine peptide, such as the tag provided in a pQE vector (QIAGEN, Inc., 9259 Eton Avenue, Chatsworth, Calif., 91311), among others, many of which are commercially available. As described in Gentz et al., 1989, Proc. Natl. Acad. Sci. USA 86:821-824, for instance, hexa-histidine provides for convenient purification of the fusion protein. Other peptide tags useful for purification include, but are not limited to, the hemagglutinin “HA” tag, which corresponds to an epitope derived from the influenza hemagglutinin protein (Wilson et al., 1984, Cell 37:767) and the “flag” tag.

[0154] In other embodiments, antibodies useful in the present invention or fragments, analogs or derivatives thereof can be conjugated to a diagnostic or detectable agent. Such antibodies can be useful for monitoring or prognosing the development or progression of a disorder as part of a clinical testing procedure, such as determining the efficacy of a particular therapy. Such diagnosis and detection can be accomplished by coupling the antibody to detectable substances including, but not limited to various enzymes, such as but not limited to horseradish peroxidase, alkaline phosphatase, beta-galactosidase, or acetylcholinesterase; prosthetic groups, such as but not limited to streptavidin/biotin and avidin/biotin; fluorescent materials, such as but not limited to, umbelliferone, fluorescein, fluorescein isothiocynate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride or phycoerythrin; luminescent materials, such as but not limited to, luminol; bioluminescent materials, such as but not limited to, luciferase, luciferin, and aequorin; radioactive materials, such as but not limited to iodine (¹³¹I, ¹²⁵I, ¹²³I, ¹²¹I), carbon (¹⁴C), sulfur (³⁵S), tritium (³H), indium (¹¹⁵In, ¹¹³In, ¹¹²In, ¹¹¹In), and technetium (⁹⁹Tc), thallium (²⁰¹Ti), gallium (⁶⁸Ga, ⁶⁷Ga), palladium (¹⁰³Pd), molybdenum (⁹⁹Mo), xenon (¹³³Xe), fluorine (¹⁸F), ¹⁵³Sm, ¹⁷⁷Lu, ¹⁵⁹Gd, ¹⁴⁹ Pm, ¹⁴⁰La, ¹⁷⁵Yb, ¹⁶⁶Ho, ⁹⁰Y, ⁴⁷Sc, ¹⁸⁶Re, ¹⁸⁸Re, ¹⁴²Pr, ¹⁰⁵Rh, ⁹⁷Ru, ⁶⁸Ge, ⁵⁷Co, ⁶⁵Zn, ⁸⁵Sr, ³²P, ¹⁵³Gd, ⁶⁹Yb, ⁵¹Cr, ⁵⁴Mn, ⁷⁵Se, ¹¹³Sn, and ¹¹⁷Tin; positron emitting metals using various positron emission tomographies, noradioactive paramagnetic metal ions, and molecules that are radiolabelled or conjugated to specific radioisotopes.

[0155] The present invention further encompasses formulations comprising antibodies that are conjugated to a therapeutic moiety. An antibody or fragment thereof may be conjugated to a therapeutic moiety such as a cytotoxin, e.g., a cytostatic or cytocidal agent, a therapeutic agent or a radioactive metal ion, e.g., alpha-emitters. A cytotoxin or cytotoxic agent includes any agent that is detrimental to cells. Therapeutic moieties include, but are not limited to, antimetabolites (e.g., methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine, 5-fluorouracil decarbazine), alkylating agents (e.g., mechlorethamine, thioepa chlorambucil, melphalan, carmustine (BCNU) and lomustine (CCNU), cyclothosphamide, busulfan, dibromomannitol, streptozotocin, mitomycin C, and cisdichlorodiamine platinum (II) (DDP) cisplatin), anthracyclines (e.g., daunorubicin (formerly daunomycin) and doxorubicin), antibiotics (e.g., dactinomycin (formerly actinomycin), bleomycin, mithramycin, and anthramycin (AMC)), Auristatin molecules (e.g., auristatin PHE, bryostatin 1, and solastatin 10; see Woyke et al., Antimicrob. Agents Chemother. 46:3802-8 (2002), Woyke et al., Antimicrob. Agents Chemother. 45:3580-4 (2001), Mohammad et al., Anticancer Drugs 12:735-40 (2001), Wall et al., Biochem. Biophys. Res. Commun. 266:76-80 (1999), Mohammad et al., Int. J. Oncol. 15:367-72 (1999), all of which are incorporated herein by reference), hormones (e.g., glucocorticoids, progestins, androgens, and estrogens), DNA-repair enzyme inhibitors (e.g., etoposide or topotecan), kinase inhibitors (e.g., compound ST1571, imatinib mesylate (Kantaijian et al., Clin Cancer Res. 8(7):2167-76 (2002)), cytotoxic agents (e.g., paclitaxel, cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicin, doxorubicin, daunorubicin, dihydroxy anthracin dione, mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone, procaine, tetracaine, lidocaine, propranolol, and puromycin and analogs or homologs thereof) and those compounds disclosed in U.S. Pat. Nos. 6,245,759, 6,399,633, 6,383,790, 6,335,156, 6,271,242, 6,242,196, 6,218,410, 6,218,372, 6,057,300, 6,034,053, 5,985,877, 5,958,769, 5,925,376, 5,922,844, 5,911,995, 5,872,223, 5,863,904, 5,840,745, 5,728,868, 5,648,239, 5,587,459), farnesyl transferase inhibitors (e.g., R115777, BMS-214662, and those disclosed by, for example, U.S. Pat. Nos. 6,458,935, 6,451,812, 6,440,974, 6,436,960, 6,432,959, 6,420,387, 6,414,145, 6,410,541, 6,410,539, 6,403,581, 6,399,615, 6,387,905, 6,372,747, 6,369,034, 6,362,188, 6,342,765, 6,342,487, 6,300,501, 6,268,363, 6,265,422, 6,248,756, 6,239,140, 6,232,338, 6,228,865, 6,228,856, 6,225,322, 6,218,406, 6,211,193, 6,187,786, 6,169,096, 6,159,984, 6,143,766, 6,133,303, 6,127,366, 6,124,465, 6,124,295, 6,103,723, 6,093,737, 6,090,948, 6,080,870, 6,077,853, 6,071,935, 6,066,738, 6,063,930, 6,054,466, 6,051,582, 6,051,574, and 6,040,305), topoisomerase inhibitors (e.g., camptothecin; irinotecan; SN-38; topotecan; 9-aminocamptothecin; GG-211 (GI 147211); DX-895 If; IST-622; rubitecan; pyrazoloacridine; XR-5000; saintopin; UCE6; UCE1022; TAN-1518A; TAN-1518B; KT6006; KT6528; ED-110; NB-506; ED-110; NB-506; and rebeccamycin); bulgarein; DNA minor groove binders such as Hoescht dye 33342 and Hoechst dye 33258; nitidine; fagaronine; epiberberine; coralyne; beta-lapachone; BC-4-1; bisphosphonates (e.g., alendronate, cimadronte, clodronate, tiludronate, etidronate, ibandronate, neridronate, olpandronate, risedronate, piridronate, pamidronate, zolendronate) HMG-CoA reductase inhibitors, (e.g., lovastatin, simvastatin, atorvastatin, pravastatin, fluvastatin, statin, cerivastatin, lescol, lupitor, rosuvastatin and atorvastatin) and pharmaceutically acceptable salts, solvates, clathrates, and prodrugs thereof. See, e.g., Rothenberg, M. L., Annals of Oncology 8:837-855(1997); and Moreau, P., et al., J. Med. Chem. 41:1631-1640(1998)), antisense oligonucleotides (e.g., those disclosed in the U.S. Pat. Nos. 6,277,832, 5,998,596, 5,885,834, 5,734,033, and 5,618,709), immunomodulators (e.g., antibodies and cytokines), antibodies, and adenosine deaminase inhibitors (e.g., Fludarabine phosphate and 2-Chlorodeoxyadenosine).

[0156] Further, an antibody or fragment thereof may be conjugated to a therapeutic moiety or drug moiety that modifies a given biological response. Therapeutic moieties or drug moieties are not to be construed as limited to classical chemical therapeutic agents. For example, the drug moiety may be a protein or polypeptide possessing a desired biological activity. Such proteins may include, for example, a toxin such as abrin, ricin A, pseudomonas exotoxin, cholera toxin, or diphtheria toxin; a protein such as tumor necrosis factor, .alpha.-interferon, .beta.-interferon, nerve growth factor, platelet derived growth factor, tissue plasminogen activator, an apoptotic agent, e.g., TNFα, TNFβ, AIM I (see, International publication No. WO 97/33899), AIM II (see, International Publication No. WO 97/34911), Fas Ligand (Takahashi et al., 1994, J. Immunol., 6:1567-1574), and VEGI (see, International publication No. WO 99/23105), a thrombotic agent or an anti-angiogenic agent, e.g., angiostatin, endostatin or a component of the coagulation pathway (e.g., tissue factor); or, a biological response modifier such as, for example, a lymphokine (e.g., interleukin-1 (“IL-1”), interleukin-2 (“IL-2”), interleukin-6 (“IL-6”), granulocyte macrophage colony stimulating factor (“GM-CSF”), and granulocyte colony stimulating factor (“G-CSF”)), a growth factor (e.g., growth hormone (“GH”)), or a coagulation agent (e.g., calcium, vitamin K, tissue factors, such as but not limited to, Hageman factor (factor XII), high-molecular-weight kininogen (HMWK), prekallikrein (PK), coagulation proteins-factors II (prothrombin), factor V, XIIa, VIII, XIIIa, XI, XIa, IX, IXa, X, phospholipid. fibrinopeptides A and B from the α and β chains of fibrinogen, fibrin monomer).

[0157] Moreover, an antibody can be conjugated to therapeutic moieties such as a radioactive metal ion, such as alph-emiters such as ²¹³Bi or macrocyclic chelators useful for conjugating radiometal ions, including but not limited to, ¹³¹In, ¹³¹LU, ¹³¹Y, ¹³¹Ho, ¹³¹Sm, to polypeptides. In certain embodiments, the macrocyclic chelator is 1,4,7,10-tetraazacyclododecane-N,N′,N″,N′″-tetraacetic acid (DOTA) which can be attached to the antibody via a linker molecule. Such linker molecules are commonly known in the art and described in Denardo et al., 1998, Clin Cancer Res. 4(10):2483-90; Peterson et al., 1999, Bioconjug. Chem. 10(4):553-7; and Zimmerman et al., 1999, Nucl. Med. Biol. 26(8):943-50, each incorporated by reference in their entireties.

[0158] Techniques for conjugating therapeutic moieties to antibodies are well known, see, e.g., Arnon et al., “Monoclonal Antibodies For Immunotargeting Of Drugs In Cancer Therapy”, in Monoclonal Antibodies And Cancer Therapy, Reisfeld et al. (eds.), pp. 243-56 (Alan R. Liss, Inc. 1985); Hellstrom et al., “Antibodies For Drug Delivery”, in Controlled Drug Delivery (2nd Ed.), Robinson et al. (eds.), pp. 623-53 (Marcel Dekker, Inc. 1987); Thorpe, “Antibody Carriers Of Cytotoxic Agents In Cancer Therapy: A Review”, in Monoclonal Antibodies 84: Biological And Clinical Applications, Pinchera et al. (eds.), pp. 475-506 (1985); “Analysis, Results, And Future Prospective Of The Therapeutic Use Of Radiolabeled Antibody In Cancer Therapy”, in Monoclonal Antibodies For Cancer Detection And Therapy, Baldwin et al. (eds.), pp. 303-16 (Academic Press 1985), and Thorpe et al., 1982, Immunol. Rev. 62:119-58.

[0159] Alternatively, an antibody can be conjugated to a second antibody to form an antibody heteroconjugate as described by Segal in U.S. Pat. No. 4,676,980, which is incorporated herein by reference in its entirety.

[0160] The therapeutic moiety or drug conjugated to an antibody or fragment thereof should be chosen to achieve the desired prophylactic or therapeutic effect(s) for a particular disorder in a subject. A clinician or other medical personnel should consider the following when deciding on which therapeutic moiety or drug to conjugate to an antibody or fragment thereof: the nature of the disease, the severity of the disease, and the condition of the subject.

[0161] Antibodies may also be attached to solid supports, which are particularly useful for immunoassays or purification of the target antigen. Such solid supports include, but are not limited to, glass, cellulose, polyacrylamide, nylon, polystyrene, polyvinyl chloride or polypropylene.

[0162] In further embodiments, the present invention encompasses formulations comprising isolated antibodies or compositions comprising antibodies, wherein said antibodies specifically bind to one or more particular antigens. In certain embodiments, the antibody of the present invention specifically binds to an antigen of respiratory syncytial virus (RSV). In other embodiments, the antibody of the present invention specifically binds to an antigen of human metapneumovirus (hMPV). In some embodiments, the antibody is a humanized antibody that specifically binds to an antigen of hMPV. In certain embodiments, the antibody of the present invention specifically binds to integrin α_vβ₃. In some embodiments, the antibody is MEDI-522 (Vitaxin®). In certain embodiments, the antibody of the present invention specifically binds to CD2. In some embodiments, the antibody is siplizumab. In certain embodiments, the antibody of the present invention specifically binds to CD19. In some embodiments, the antibody is MT-103. In further embodiments, the antibody of the present invention specifically binds to EphA2. In some embodiments, the antibody is human or humanized EA2 or EA5. In certain embodiments, the antibody of the present invention specifically binds to EphA4. In some embodiments, the antibody is a humanized antibody that specifically binds to EphA4. In certain embodiments, the antibody of the present invention specifically binds to IL-9. In some embodiments, the antibody is a human or humanized antibody that specifically binds to IL-9. In some embodiments, the antibody is MEDI-528.

[0163] In some embodiments, the antibody is not palivizumab. In some embodiments, the antibody is not MEDI-522 (Vitaxin®). In some embodiments, the antibody is not siplizumab. In some embodiments, the antibody is not MT-103. In some embodiments, the antibody is not human or humanized EA2 or EA5. In some embodiments, the antibody is not MEDI-528.

[0164] The antibodies useful in the present invention may be high potency antibodies. High potency antibodies can be produced by genetically engineering appropriate antibody gene sequences and expressing the antibody sequences in a suitable host. The antibodies produced can be screened to identify antibodies with, e.g., high k_on values in a BIAcore assay.

[0165] In certain embodiments, the antibodies useful in the present invention have a high binding affinity for one or more antigens. In a specific embodiment, the antibodies of the present invention have an association rate constant or k_on rate (antibody (Ab)+antigen (Ag)^kno→Ab—Ag) of at least 10⁵ M⁻¹s⁻¹, at least 5×10⁵ M⁻¹s⁻¹, at least 10⁶ M⁻¹s⁻¹, at least 5×10⁶ M⁻¹s⁻¹, at least 10⁷ M⁻¹s⁻¹, at least 5×10⁷ M⁻¹s⁻¹, or at least 10⁸ M⁻¹s⁻¹. In a preferred embodiment, the antibodies of the present invention have a k_on of at least 2×10⁵ M⁻¹s⁻¹, at least 5×10⁵ M⁻¹s⁻¹, at least 10⁶ M⁻¹s⁻¹, at least 5×10⁶ M⁻¹s⁻¹, at least 10⁷ M⁻¹s⁻¹, at least 5×10⁷ M⁻¹s⁻¹, or at least 10⁸ M⁻¹s⁻¹.

[0166] In another embodiment, the antibodies of the present invention have a k_off rate (antibody (Ab)+antigen) of less than 10⁻¹ s⁻¹, less than 5×10⁻¹ s⁻¹, less than 10⁻² s⁻¹, less than 5 ×10⁻² s⁻¹, less than 10⁻³ s⁻¹, less than 5×10⁻³ s⁻¹, less than 10⁻⁴ s⁻¹, less than 5×10⁻⁴ s⁻¹, less than 10⁻⁵ s⁻¹, less than 5×10⁻⁵ s⁻¹, less than 10⁻⁶ s⁻¹, less than 5×10⁻⁶ s⁻¹, less than 10⁻⁷ s⁻¹, less than 5×10⁻⁷ s⁻¹, less than 10⁻⁸ s⁻¹, less than 5×10⁻⁸ s⁻¹, less than 10⁻⁹ s⁻¹, less than 5×10⁻⁹ s⁻¹, or less than 10⁻¹⁰ s⁻¹. In a preferred embodiment, the antibodies of the present invention have a k_on of less than 5×10⁻⁴ s⁻¹, less than 10⁻⁵ s⁻¹, less than 5×10⁻⁵ s⁻¹, less than 10⁻⁶ s⁻¹, less than 5×10⁻⁶ s⁻¹, less than 10⁻⁷ s⁻¹, less than 5×10⁻⁷ s⁻¹, less than 10⁻⁸ s⁻¹, less than 5×10⁻⁸ s⁻¹, less than 10⁻⁹ s⁻¹, less than 5×10⁻⁹ s⁻¹, or less than 10⁻¹⁰ s⁻¹.

[0167] In certain embodiments, the antibodies of the present invention have an affinity constant or K_a (k_on/k_off) of at least 10² M⁻¹, at least 5×10² M⁻¹, at least 10³ M⁻¹, at least 5×10³ M⁻¹, at least 10⁴ M⁻¹, at least 5×10⁴ M⁻¹, at least 10⁵ M⁻¹, at least 5×10⁵ M⁻¹, at least 10⁶ M⁻¹, at least 5×10⁶ M⁻¹, at least 10⁷ M⁻¹, at least 5×10⁷ M⁻¹, at least 10⁸ M⁻¹, at least 5×10⁸ M⁻¹, at least 10⁹ M⁻¹, at least 5×10⁹ M⁻¹, at least 10¹⁰ M⁻¹, at least 5×10¹⁰ M⁻¹, at least 10¹¹ M⁻¹, at least 5×10¹¹ M⁻¹, at least 10¹² M⁻¹, at least 5×10¹² M⁻¹, at least 10¹³ M⁻¹, at least 5×10¹³ M⁻¹, at least 10¹⁴ M⁻¹, at least 5×10¹⁴ M⁻¹, at least 10¹⁵ M⁻¹, or at least 5×10¹⁵ M⁻¹. The present invention also provides formulations comprising one or more antibodies which immunospecifically bind to an antigen with an affinity constant of at least 2×10⁸ M⁻¹, at least 2.5×10⁸ M⁻¹, at least 5×10⁸ M⁻¹, at least 10⁹ M⁻¹, at least 5×10⁹ M⁻¹, at least 10¹⁰ M⁻¹, at least 5×10⁻¹ M, at least 10¹¹ M⁻¹, at least 5×10¹¹ M⁻¹, at least 10¹² M⁻¹, at least 5×10¹² M, at least 10¹³ M⁻¹, at least 5×10¹³ M⁻¹, at least 10¹⁴ M⁻¹, at least 5×10¹⁴ M⁻¹, at least 10¹⁵ M⁻¹, or at least 5×10¹⁵ M⁻¹.

[0168] In yet another embodiment, the antibodies useful in the present invention have a dissociation constant or K_d (k_off/k_on) of less than 10⁻² M, less than 5×10⁻² M, less than 10⁻³ M, less than 5×10⁻³ M, less than 10⁻⁴ M, less than 5×10⁻⁴ M, less than 10⁻⁵ M, less than 5×10⁻⁵ M, less than 10⁻⁶ M, less than 5×10⁻⁶ M, less than 10⁻⁷ M, less than 5×10⁻⁷ M, less than 10⁻⁸ M, less than 5×10⁻⁸ M, less than 10⁻⁹ M, less than 5×10⁻⁹ M, less than 10⁻¹⁰ M, less than 5×10⁻¹⁰ M, less than 10⁻¹¹ M, less than 5×10⁻¹¹ M, less than 10⁻¹² M, less than 5×10⁻¹² M, less than 10⁻¹³ M, less than 5×10⁻¹³ M, less than 10⁻¹⁴ M, less than 5×10⁻¹⁴ M, less than 10⁻¹⁵ M, or less than 5×10⁻¹⁵ M.

[0169] In certain embodiments, the antibodies useful in the present invention have a median effective concentration (EC₅₀) of less than 0.01 nM, less than 0.025 nM, less than 0.05 nM, less than 0.1 nM, less than 0.25 nM, less than 0.5 nM, less than 0.75 nM, less than 1 nM, less than 1.25 nM, less than 1.5 nM, less than 1.75 nM, or less than 2 nM, in an in vitro microneutralization assay. The median effective concentration is the concentration of antibody or antibody fragments that neutralizes 50% of an antigen in an in vitro microneutralization assay. In a preferred embodiment, the antibodies of the present invention have an EC₅₀ of less than 0.01 nM, less than 0.025 nM, less than 0.05 nM, less than 0.1 nM, less than 0.25 nM, less than 0.5 nM, less than 0.75 nM, less than 1 nM, less than 1.25 nM, less than 1.5 nM, less than 1.75 nM, or less than 2 nM, in an in vitro microneutralization assay.

[0170] The present invention also provides antibodies that immunospecifically bind to an antigen of interest, the antibodies comprising derivatives of the VH domains, VH CDRs, VL domains, and VL CDRs described herein that immunospecifically bind to antigens of interest. Standard techniques known to those of skill in the art can be used to introduce mutations in the nucleotide sequence encoding a molecule of the invention, including, for example, site-directed mutagenesis and PCR-mediated mutagenesis which results in amino acid substitutions. Preferably, the derivatives include less than 25 amino acid substitutions, less than 20 amino acid substitutions, less than 15 amino acid substitutions, less than 10 amino acid substitutions, less than 5 amino acid substitutions, less than 4 amino acid substitutions, less than 3 amino acid substitutions, or less than 2 amino acid substitutions relative to the original molecule. In a preferred embodiment, the derivatives have conservative amino acid substitutions are made at one or more predicted non-essential amino acid residues. A “conservative amino acid substitution” is one in which the amino acid residue is replaced with an amino acid residue having a side chain with a similar charge. Families of amino acid residues having side chains with similar charges have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Alternatively, mutations can be introduced randomly along all or part of the coding sequence, such as by saturation mutagenesis, and the resultant mutants can be screened for biological activity to identify mutants that retain activity. Following mutagenesis, the encoded protein can be expressed and the activity of the protein can be determined.

[0171] It should be recognized that antibodies that immunospecifically bind to a RSV antigen are known in the art. For example, palivizumab is a humanized monoclonal antibody presently used for the prevention of RSV infection in pediatric patients. The present invention provides formulations of antibodies that immunospecifically bind to one or more RSV antigens. Preferably, the antibodies useful in the invention immunospecifically bind to one or more RSV antigens regardless of the strain of RSV. The present invention also provides antibodies that differentially or preferentially bind to RSV antigens from one strain of RSV versus another RSV strain. In a specific embodiment, the formulations comprise antibodies that immunospecifically bind to the RSV F glycoprotein, G glycoprotein or SH protein. In a preferred embodiment, the formulations comprise antibodies that immunospecifically bind to the RSV F glycoprotein. In another preferred embodiment, the formulations comprise antibodies that bind to the A, B, or C antigenic sites of the RSV F glycoprotein.

[0172] The formulations of the invention comprise antibodies that immunospecifically bind to a RSV antigen and have a dissociation constant (K_D) of less than 3000 pM, less than 2500 pM, less than 2000 pM, less than 1500 pM, less than 1000 pM, less than 750 pM, less than 500 pM, less than 250 pM, less than 200 pM, less than 150 pM, less than 100 pM, less than 75 pM as assessed using an described herein or known to one of skill in the art (e.g., a BIAcore assay). In a specific embodiment, formulations of the invention comprise antibodies that immunospecifically bind to a RSV antigen and have a dissociation constant (K_D) of between 25 to 3400 pM, 25 to 3000 pM, 25 to 2500 pM, 25 to 2000 pM, 25 to 1500 pM, 25 to 1000 pM, 25 to 750 pM, 25 to 500 pM, 25 to 250 pM, 25 to 100 pM, 25 to 75 pM, 25 to 50 pM as assessed using an described herein or known to one of skill in the art (e.g., a BIAcore assay). In another embodiment, formulations of the invention comprise antibodies that immunospecifically bind to a RSV antigen and have a dissociation constant (K_D) of 500 pM, preferably 100 pM, more preferably 75 pM and most preferably 50 pM as assessed using an described herein or known to one of skill in the art (e.g., a BIAcore assay).

[0173] The present invention provides formulations that comprise antibodies that have a median inhibitory concentration (IC₅₀) of less than 5 nM, less than 4 nM, less than 3 nM, less than 2 nM, less than 1.75 nM, less than 1.5 nM, less than 1.25 nM, less than 1 nM, less than 0.75 nM, less than 0.5 nM, less than 0.25 nM, less than 0.1 nM, less than 0.05 nM, less than 0.025 nM, or less than 0.01 nM, in an in vitro microneutralization assay. The IC₅₀ is the concentration of antibody that neutralizes 50% of the RSV in an in vitro microneutralization assay. In a preferred embodiment, antibody of the invention has an IC₅₀ of less than 5 nM, less than 4 nM, less than 3 nM, less than 2 nM, less than 1.75 nM, less than 1.5 nM, less than 1.25 nM, less than 1 nM, less than 0.75 nM, less than 0.5 nM, less than 0.25 nM, less than 0.1 nM, less than 0.05 nM, less than 0.025 nM, or less than 0.01 nM, in an in vitro microneutralization assay.

[0174] In a specific embodiment, the formulations of the invention comprise an antibody that has approximately 20-fold, 25-fold, 30-fold, 35-fold, 40-fold, 45-fold, 50-fold, 55-fold, 60-fold, 65-fold, 70-fold, 75-fold, 80-fold, 90-fold, 100-fold or higher affinity for a RSV F antigen than palivizumab or an antibody-binding fragment thereof as assessed by an assay known in the art or described herein (e.g., a BIAcore assay). In another embodiment, formulations of the invention comprise antibodies that have an approximately 1-fold, 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, or more higher K_a than palivizumab or an antigen-binding fragment thereof as assessed by an assay known in the art or described herein. In another embodiment, a formulation of the invention comprises an antibody that is approximately 1-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 11-fold 12-fold, 13-fold, 14-fold, 15-fold, 16-fold, 17-fold, 18-fold, 19-fold, or 20-fold or more potent than palivizumab or an antigen-binding fragment thereof in an in vitro microneutralization assay such as described herein. The amino acid sequence of palivizumab is disclosed, e.g., in Johnson et al., 1997, J. Infectious Disease 176:1215-1224, and U.S. Pat. No. 5,824,307, each of which is incorporated herein by reference in its entirety. In a specific embodiment, a formulation of the invention comprise an antibody that is not palivizumab or a fragment of palivizumab or an antigen-binding fragment of palivizumab, e.g., is not an antibody comprising a VH domain of SEQ ID NO:7 and/or a VL domain of SEQ ID NO:8.

[0175] The present invention provides antibodies that immunospecifically bind to one or more RSV antigens, said antibodies comprising the amino acid sequence of palivizumab with one or more amino acid residue substitutions in the variable light (VL) domain and/or variable heavy (VH) domain depicted in FIG. 3. The present invention also provides antibodies that immunospecifically bind to one or more RSV antigens, said antibodies comprising the amino acid sequence of palivizumab with one or more amino acid residue substitutions in one or more VL CDRs and/or one or more VH CDRs. In a specific embodiment, an antibody comprises the amino acid sequence of palivizumab with one or more amino acid residue substitutions of the amino acid residues indicated in bold face and underlining in Table 1. In another embodiment, an antibody comprises the amino sequence of palivizumab with one or more amino acid residue substitutions of the amino acid residues indicated in bold face and underlining in Table 1 and one or more amino acid residue substitutions of the framework regions of the variable domains of palivizumab (e.g., mutations in framework region 4 of the heavy and/or light variable domains). In accordance with these embodiments, the amino acid residue substitutions can be conservative or non-conservative. The antibody generated by introducing substitutions in the VH domain, VH CDRs, VL domain and/or VL CDRs of palivizumab can be tested in vitro and in vivo, for example, for its ability to bind to RSV F antigen, for its ability to neutralize RSV, or for its ability to prevent, treat or ameliorate an upper and/or lower respiratory tract RSV infection, otitis media, or a symptom or respiratory condition relating thereto (including, but not limited to, asthma, wheezing, RAD, or a combination thereof).

TABLE 1
CDR Sequences of palivizumab
	CDR	Sequence	SEQ ID NO:
	VH1	TSGMSVG	1
	VH2	DIWWDDKKDYNPSLKS	2
	VH3	SMITNWYFDV	3
	VL1	KCQLSVGYMH	4
	VL2	DTSKLAS	5
	VL3	FQGSGYPFT	6
	* Bold faced & underlined amino acid residues are preferred residues which should be substituted.

[0176] The formulations of the present invention also comprise those antibodies and antigen-binding fragments of the antibodies referenced in Table 2 and the Examples Section of the application. In a specific embodiment, a formulation of the present invention comprises antibody AFFF, P12f2, P12f4, P11d4, Ale9, A12a6, A13c4, A17d4, A4B4, A8c7, 1X-493L1FR, H3-3F4, M3H9, Y10H6, DG, AFFF(1), 6H8, L1-7E5, L2-15B10, A13a11, A1h5, A4B4(1), A4B4L1FR-S28R (motavizumab), A4B4-F52S, A17d4(1), A3e2, A14a4, A16b4, A17b5, A17f5, or A17h4. In another embodiment, a formulation of the present invention comprises an antigen-binding fragment (e.g., a Fab fragment of) AFFF, P12f2, P12f4, P11d4, Ale9, A12a6, A13c4, A17d4, A4B4, A8c7, 1X-493L1FR, H3-3F4, M3H9, Y10H6, DG, AFFF(1), 6H8, L1-7E5, L2-15B10, A13a11, A1h5, A4B4(1), A4B4L1FR-S28R (motavizumab), A4B4-F52S, A17d4(1), A3e2, A14a4, A16b4, A17b5, A17f5, or A17h4. In a preferred embodiment, a formulation of the present invention comprises antibody A4B4L1FR-S28R or an antigen-binding fragment thereof.

[0177] In some embodiments, AFFF, P12f2, P12f4, P11d4, Ale9, A12a6, A13c4, A17d4, A4B4, A8c7, 1X-493L1FR, H3-3F4, M3H9, Y10H6, DG, AFFF(1), 6H8, L1-7E5, L2-15B10, A13a11, A1h5, A4B4(1), A4B4L1FR-S28R, A4B4-F52S, A17d4(1), A3e2, A14a4, A16b4, A17b5, A17f5, and/or A17h4 comprise the framework region and constant regions of palivizumab (see FIG. 3). In preferred embodiments, AFFF, P12f2, P12f4, P11d4, Ale9, A12a6, A13c4, A17d4, A4B4, A8c7, 1X-493L1FR, H3-3F4, M3H9, Y10H6, DG, AFFF(1), 6H8, L1-7E5, L2-15B10, A13a11, A1h5, A4B4(1), A4B4L1FR-S28R (motavizumab), A4B4-F52S, A17d4(1), A3e2, A14a4, A16b4, A17b5, A17f5, and/or A17h4 comprise the framework region and constant regions of palivizumab with the exception that there is an amino acid substitution of an A105Q in the heavy chain framework 4 (FR4) (numbering used herein according to Kabat et al. (1991). Sequences of proteins of immunological interest. (U.S. Department of Health and Human Services, Washington, D.C.) 5^th ed.) (“Kabat numbering”) (i.e., position 112 in SEQ ID NO:7 (palivizumab VH domain)) and an L104V in the light chain FR4 (i.e., position 103 in SEQ ID NO:8 (palivizumab VL domain)). An example of antibodies comprising a framework with these VH and VL single mutations is shown in FIG. 4 (1X-493L1FR) and in FIG. 5 (A4B4L 1 FR-S28R).

[0178] In a specific embodiment, the present invention provides one or more antibodies that immunospecifically bind to one or more RSV F antigens, said antibodies comprising a VH chain and/or VL chain having the amino acid sequence of a VH chain and/or VL chain of AFFF, P12f2, P12f4, P11d4, Ale9, A12a6, A13c4, A17d4, A4B4, A8c7, 1X-493L1FR, H3-3F4, M3H9, Y10H6, DG, AFFF(1), 6H8, L1-7E5, L2-15B10, A13a11, A1h5, A4B4(1), A4B4L1FR-S28R, A4B4-F52S, A17d4(1), A3e2, A14a4, A16b4, A17b5, A17f5, or A17h4. In a preferred embodiment, an antibody of the invention immunospecifically binds to a RSV F antigen, and said antibody comprises a VH chain and/or a VL chain having the amino acid sequence of the VH and/or VL chain of A4B4L1FR-S28 (VH chain, SEQ ID NO:254; VL chain SEQ ID NO:255). In another embodiment, the present invention provides one or more antibodies that immunospecifically bind to one or more RSV antigens, said antibodies comprising a VH domain and/or VL domain having the amino acid sequence of a VH domain and/or VL domain of AFFF, P12f2, P12f4, P11d4, Ale9, A12a6, A13c4, A17d4, A4B4, A8c7, 1X-493L1FR, H3-3F4, M3H9, Y10H6, DG, AFFF(1), 6H8, L1-7E5, L2-15B10, A13a11, A1h5, A4B4(1), A4B4L1FR-S28R, A4B4-F52S, A17d4(1), A3e2, A14a4, A16b4, A17b5, A17f5, or A17h4. In a preferred embodiment, an antibody of the invention immunospecifically binds to a RSV F antigen, and said antibody comprises a VH domain and/or VL domain having the amino acid sequence of the VH domain and/or VL domain of A4B4L1FR-S28R (VH domain, SEQ ID NO:48; VL domain, SEQ ID NO:11).

[0179] In another embodiment, the present invention provides antibodies that immunospecifically bind to one or more RSV antigens, said antibodies comprising one, two, three, or more CDRs having the amino acid sequence of one, two, three, or more CDRs of AFFF, P12f2, P12f4, P11d4, Ale9, A 12a6, A13c4, A17d4, A4B4, A8c7, 1X-493L1FR, H3-3F4, M3H9, Y10H6, DG, AFFF(1), 6H8, L1-7E5, L2-15B10, A13a11, A1h5, A4B4(1), A4B4 L1FR-S28R (MEDI-524, motavizumab), A4B4-F52S, A17d4(1), A3e2, A14a4, A16b4, A17b5, A17f5, or A17h4. In a preferred embodiment, a formulation of the present invention comprises an antibody that immunospecifically binds to a RSV antigen, and said antibody comprises one, two, three, or more CDRs having the amino acid sequence of one, two, three, or more CDRs of A4B4L1FR-S28R. In yet another embodiment, the formulation of the present invention comprises an antibody that immunospecifically binds to one or more RSV F antigens, said antibodies comprising a combination of VH CDRs and/or VL CDRs having the amino acid sequence of VH CDRs and/or VL CDRs of AFFF, P12f2, P12f4, P11d4, Ale9, A12a6, A13c4, A17d4, A4B4, A8c7, 1X-493L1FR, H3-3F4, M3H9, Y10H6, DG, AFFF(1), 6H8, L1-7E5, L2-15B10, A13a11, A1h5, A4B4(1), A4B4L1FR-S28R (motavizumab), A4B4-F52S, A17d4(1), A3e2, A14a4, A16b4, A17b5, A17f5, and/or A17h4. In a preferred embodiment, a formulation of the present invention comprises an antibody that immunospecifically binds to a RSV F antigen and said antibody comprises a combination of VH CDRs and/or VL CDRs having the amino acid sequence of the VH CDRs and/or VL CDRs of A4B4L1FR-S28R.

[0180] The present invention provides antibodies that immunospecifically bind to one or more RSV antigens (e.g., RSV F antigen), said antibodies comprising a variable heavy (“VH”) chain having an amino acid sequence of any one of the VH chains listed in Table 2. In certain embodiments, the antibody is not palivizumab and/or the VH chain is not the VH chain of palivizumab.

[0181] The invention also provides antibodies that immunospecifically bind to one or more RSV antigens (e.g., RSV F antigen), said antibodies comprising a VH domain having an amino acid sequence of any one of the VH domains listed in Table 2. In certain embodiments of the invention, the antibody is not palivizumab and/or the VH domain is not the VH domain of palivizumab.

[0182] The present invention also provides antibodies that immunospecifically bind to one or more RSV antigens, said antibodies comprising a VH complementarity determining region (“CDR”) (e.g., VH CDR1, VH CDR2, and/or VH CDR3) having an amino acid sequence of any of the VH CDRs listed in Table 2 and/or Tables 3A-3C. In certain embodiments of the invention, an antibody comprising a VH CDR having an amino acid of any of one of the VH CDRs listed in Table 2 and/or Tables 3A-3C is not palivizumab. In some embodiments, the antibody or binding fragment thereof comprises one, two or three of the VH CDRs listed in Table 2 and/or Tables 3A-3C.

TABLE 2
Antibodies & Fragments Thereof
Antibody	VH	VH				VL	VL
Name	Chain	Domain	VH CDR1	VH CDR2	VH CDR3	Chain	Domain	VL CDR1	VL CDR2	VL CDR3
**pali-	SEQ ID	SEQ ID	TSGMSVG	DIWWDDKKDYN	SMITNWYFDV	SEQ ID	SEQ ID	KCQLSVGYMH	DTSKLAS	FQGSGYPFT
vizumab	NO:208	NO:7	(SEQ ID	PSLKS	(SEQ ID	NO:209	NO:8	(SEQ ID	(SEQ ID	(SEQ ID
			NO:1)		NO:3)			NO:4)	NO:5)	NO:6)
				(SEQ ID NO:2)
***AFFF	SEQ ID	SEQ ID	TAGMSVG	DIWWDDKKDYN	SMITNFYFDV	SEQ ID	SEQ ID	SASSSVGYMH	DTFKLAS	FQFSGYPFT
	NO:210	NO:9	(SEQ ID	PSLKS	(SEQ ID	NO:211	NO:13	(SEQ ID	(SEQ ID	(SEQ ID
			NO:10)	(SEQ ID NO:2)	NO:12)			NO:14)	NO:15)	NO:16)
***P12f2	SEQ ID	SEQ ID	TPGMSVG	DIWWDDKKHYN	DMIFNFYFDV	SEQ ID	SEQ ID	SLSSRVGYMH	DTFYLSS	FQGSGYPFT
	NO:212	NO:17	(SEQ ID	PSLKD	(SEQ ID	NO:213	NO:21	(SEQ ID	(SEQ ID	(SEQ ID
			NO:18)	(SEQ ID NO:19)	NO:20)			NO:22)	NO:23)	NO:6)
***P12f4	SEQ ID	SEQ ID	TPGMSVG	DIWWDGKKHYN	DMIFNFYFDV	SEQ ID	SEQ ID	SLSSRVGYMH	DTRGLPS	FQGSGYPFT
	NO:214	NO:24	(SEQ ID	PSLKD	(SEQ ID	NO:215	NO:26	(SEQ ID	(SEQ ID	(SEQ ID
			NO:18)	(SEQ ID NO:25)	NO:20)			NO:22)	NO:27)	NO:6)
***P11d4	SEQ ID	SEQ ID	TPGMSVG	DIWWDGKKHYN	DMIFNWYFDV	SEQ ID	SEQ ID	SPSSRVGYMH	DTMRLAS	FQGSGYPFT
	NO:216	NO:28	(SEQ ID	PSLKD	(SEQ ID	NO:217	NO:30	(SEQ ID	(SEQ ID	(SEQ ID
			NO:18)	(SEQ ID NO:25)	NO:29)			NO:31)	NO:32)	NO:6)
***Ale9	SEQ ID	SEQ ID	TAGMSVG	DIWWDGKKHYN	DMIFNWYFDV	SEQ ID	SEQ ID	SLSSRVGYMH	DTFKLSS	FQGSGYPFT
	NO:218	NO:33	(SEQ ID	PSLKD	(SEQ ID	NO:219	NO:34	(SEQ ID	(SEQ ID	(SEQ ID
			NO:10)	(SEQ ID NO:25)	NO:29)			NO:22)	NO:35)	NO:6)
***A12a6	SEQ ID	SEQ ID	TAGMSVG	DIWWDGKKDYN	DMIFNFYFDV	SEQ ID	SEQ ID	SASSRVGYMH	DTFKLSS	FQGSGYPFT
	NO:220	NO:36	(SEQ ID	PSLKD	(SEQ ID	NO:221	NO:38	(SEQ ID	(SEQ ID	(SEQ ID
			NO:10)	(SEQ ID NO:37)	NO:20)			NO:39)	NO:35)	NO:6)
***A13c4	SEQ ID	SEQ ID	TAGMSVG	DIWWDGKKSYN	DMIFNFYFDV	SEQ ID	SEQ ID	SLSSRVGYMH	DTMYQSS	FQGSGYPFT
	NO:222	NO:40	(SEQ ID	PSLKD	(SEQ ID	NO:223	NO:42	(SEQ ID	(SEQ ID	(SEQ ID
			NO:10)	(SEQ ID NO:41)	NO:20)			NO:22)	NO:43)	NO:6)
***A17d4	SEQ ID	SEQ ID	TAGMSVG	DIWWDDKKSYN	DMIFNFYFDV	SEQ ID	SEQ ID	LPSSRVGYMH	DTMYQSS	FQGSGYPFT
	NO:224	NO:44	(SEQ ID	PSLKD	(SEQ ID	NO:225	NO:46	(SEQ ID	(SEQ ID	(SEQ ID
			NO:10)	(SEQ ID NO:45)	NO:20)			NO:47)	NO:43)	NO:6)
***A4B4	SEQ ID	SEQ ID	TAGMSVG	DIWWDDKKHYN	DMIFNFYFDV	SEQ ID	SEQ ID	SASSRVGYMH	DTFFLDS	FQGSGYPFT
	NO:226	NO:48	(SEQ ID	PSLKD	(SEQ ID	NO:227	NO:49	(SEQ ID	(SEQ ID	(SEQ ID
			NO:10)	(SEQ ID NO:19)	NO:20)			NO:39)	NO:50)	NO:6)
****A8c7	SEQ ID	SEQ ID	TAGMSVG	DIWWDDKKSYN	DMIFNWYFDV	SEQ ID	SEQ ID	SPSSRVGYMH	DTRYQSS	FQGSGYPFT
	NO:228	NO:51	(SEQ ID	PSLKD	(SEQ ID	NO:229	NO:52	(SEQ ID	(SEQ ID	(SEQ ID
			NO:10)	(SEQ ID NO:45)	NO:29)			NO:31)	NO:53)	NO:6)
*1X-	SEQ ID	SEQ ID	TSGMSVG	DIWWDDKKDYN	SMITNWYFDV	SEQ ID	SEQ ID	SASSSVGYMH	DTSKLAS	FQGSGYPFT
493L1FR	NO:230	NO:343	(SEQ ID	PSLKS	(SEQ ID	NO:231	NO:54	(SEQ ID	(SEQ ID	(SEQ ID
			NO:1)	(SEQ ID NO:2)	NO:3)			NO:14)	NO:5)	NO:6)
*H3-3F4	SEQ ID	SEQ ID	TAGMSVG	DIWWDDKKDYN	DMIFNWYFDV	SEQ ID	SEQ ID	SASSSVGYMH	DTFKLAS	FQGSGYPFT
	NO:232	NO:55	(SEQ ID	PSLKS	(SEQ ID	NO:233	NO:56	(SEQ ID	(SEQ ID	(SEQ ID
			NO:10)	(SEQ ID NO:2)	NO:29)			NO:14)	NO:15)	NO:6)
*M3H9	SEQ ID	SEQ ID	TAGMSVG	DIWWDDKKDYN	DMIFNWYFDV	SEQ ID	SEQ ID	SASSSVGYMH	DTYKQTS	FQGSGYPFT
	NO:234	NO:55	(SEQ ID	PSLKS	(SEQ ID	NO:235	NO:70	(SEQ ID	(SEQ ID	(SEQ ID
			NO:10)	(SEQ ID NO:2)	NO:29)			NO:14)	NO:57)	NO:6)
*Y10H6	SEQ ID	SEQ ID	TAGMSVG	DIWWDDKKDYN	DMIFNWYFDV	SEQ ID	SEQ ID	SASSSVGYMH	DTRYLSS	FQGSGYPFT
	NO:236	NO:55	(SEQ ID	PSLKS	(SEQ ID	NO:237	NO:58	(SEQ ID	(SEQ ID	(SEQ ID
			NO:10)	(SEQ ID NO:2)	NO:29)			NO:14)	NO:59)	NO:6)
*DG	SEQ ID	SEQ ID	TAGMSVG	DIWWDDKKDYN	DMITNFYFDV	SEQ ID	SEQ ID	SASSSVGYMH	DTFKLAS	FQGSGYPFT
(aka	NO:238	NO:78	(SEQ ID	PSLKS	(SEQ ID	NO:239	NO:56	(SEQ ID	(SEQ ID	(SEQ ID
D95/G93)			NO:10)	(SEQ ID NO:2)	NO:79)			NO:14)	NO:15)	NO:6)
AFFF(1)	SEQ ID	SEQ ID	TAGMSVG	DIWWDDKKDYN	SMITNFYFDV	SEQ ID	SEQ ID	SASSSVGYMH	DTFKLAS	FQGSFYPFT
	NO:240	NO:9	(SEQ ID	PSLKS	(SEQ ID	NO:241	NO:60	(SEQ ID	(SEQ ID	(SEQ ID
			NO:10)	(SEQ ID NO:2)	NO:12)			NO:14)	NO:15)	NO:61)
*6H8	SEQ ID	SEQ ID	TAGMSVG	DIWWDDKKDYN	DMITNFYFDV	SEQ ID	SEQ ID	SASSSVGYMH	DTFKLTS	FQGSGYPFT
	NO:242	NO:78	(SEQ ID	PSLKS	(SEQ ID	NO:243	NO:62	(SEQ ID	(SEQ ID	(SEQ ID
			NO:10)	(SEQ ID NO:2)	NO:79)			NO:14)	NO:63)	NO:6)
*L1-7E5	SEQ ID	SEQ ID	TAGMSVG	DIWWDDKKDYN	DMITNFYFDV	SEQ ID	SEQ ID	SASSRVGYMH	DTFKLAS	FQGSGYPFT
	NO:244	NO:78	(SEQ ID	PSLKS	(SEQ ID	NO:245	NO:64	(SEQ ID	(SEQ ID	(SEQ ID
			NO:10)	(SEQ ID NO:2)	NO:79)			NO:39)	NO:15)	NO:6)
*L-15B10	SEQ ID	SEQ ID	TAGMSVG	DIWWDDKKDYN	DMITNFYFDV	SEQ ID	SEQ ID	SASSSVGYMH	DTFRLAS	FQGSGYPFT
	NO:246	NO:78	(SEQ ID	PSLKS	(SEQ ID	NO:247	NO:65	(SEQ ID	(SEQ ID	(SEQ ID
			NO:10)	(SEQ ID NO:2)	NO:79)			NO:14)	NO:66)	NO:6)
*A13a11	SEQ ID	SEQ ID	TAGMSVG	DIWWDDKKHYN	DMIFNWYFDV	SEQ ID	SEQ ID	SPSSRVGYMH	DTYRHSS	FQGSGYPFT
	NO:248	NO:67	(SEQ ID	PSLKD	(SEQ ID	NO:249	NO:68	(SEQ ID	(SEQ ID	(SEQ ID
			NO:10)	(SEQ ID NO:19)	NO:29)			NO:31)	NO:69)	NO:6)
*A1h5	SEQ ID	SEQ ID	TAGMSVG	DIWWDGKKHYN	DMIFNWYFDV	SEQ ID	SEQ ID	SLSSSVGYMH	DTFFHRS	FQGSGYPFT
	NO:250	NO:33	(SEQ ID	PSLKD	(SEQ ID	NO:251	NO:71	(SEQ ID	(SEQ ID	(SEQ ID
			NO:10)	(SEQ ID NO:25)	NO:29)			NO:72)	NO:73)	NO:6)
A4B4(1)	SEQ ID	SEQ ID	TAGMSVG	DIWWDDKKHYN	DMIFNFYFDV	SEQ ID	SEQ ID	SASSRVGYMH	DTLLLDS	FQGSGYPFT
	NO:252	NO:48	(SEQ ID	PSLKD	(SEQ ID	NO:253	NO:74	(SEQ ID	(SEQ ID	(SEQ ID
			NO:10)	(SEQ ID NO:19)	NO:20)			NO:39)	NO:75)	NO:6)
***A4B4L	SEQ ID	SEQ ID	TAGMSVG	DIWWDDKKHYN	DMIFNFYFDV	SEQ ID	SEQ ID	SASSRVGYMH	DTSKLAS	FQGSGYPFT
1FR-S28R	NO:254	NO:48	(SEQ ID	PSLKD	(SEQ ID	NO:255	NO:11	(SEQ ID	(SEQ ID	(SEQ ID
(aka			NO:10)	(SEQ ID NO:19)	NO:20)			NO:39)	NO:5)	NO:6)
mota-
vizumab)
***A4B4-	SEQ ID	SEQ ID	TAGMSVG	DIWWDDKKHYN	DMIFNFYFDV	SEQ ID	SEQ ID	SASSRVGYMH	DTSFLDS	FQGSGYPFT
F52S	NO:256	NO:48	(SEQ ID	PSLKD	(SEQ ID	NO:257	NO:76	(SEQ ID	(SEQ ID	(SEQ ID
			NO:10)	(SEQ ID NO:19)	NO:20)			NO:39)	NO:77)	NO:6)
***A17d4	SEQ ID	SEQ ID	TAGMSVG	DIWWDGKKSYN	DMIFNFYFDV	SEQ ID	SEQ ID	LPSSRVGYMH	DTMYQSS	FQGSGYPFT
(1)	NO:222	NO:40	(SEQ ID	PSLKD	(SEQ ID	NO:225	NO:46	(SEQ ID	(SEQ ID	(SEQ ID
			NO:10)	(SEQ ID NO:41)	NO:20)			NO:47)	NO:43)	NO:6)
***A3e2	SEQ ID	SEQ ID	TAGMSVG	DIWWGDKGHYN	DMIFNWYFDV	SEQ ID	SEQ ID	SASSSVGYMH	DTFYLHS	FQGSGYPFT
	NO:303	NO:304	(SEQ ID	PSLKD	(SEQ ID	NO:306	NO:307	(SEQ ID	(SEQ ID	(SEQ ID
			NO:10)	(SEQ ID NO:	NO:29)			NO:14)	NO:308	NO:6
				305)
***A14a4	SEQ ID	SEQ ID	TAGMSVG	DIWWDDKKSYN	DMITNWYFDV	SEQ ID	SEQ ID	LLSSRVGYMH	DTYYQTS	FQGSGYPFT
	NO:309	NO:310	(SEQ ID	PSLKD	(SEQ ID	NO:312	NO:313	(SEQ ID	(SEQ ID	(SEQ ID
			NO:10)	(SEQ ID NO:45)	NO:311)			NO:314)	NO:315)	NO:6)
***A16b4	SEQ ID	SEQ ID	TAGMSVG	DIWWDDKKHYN	DMIFNWYFDV	SEQ ID	SEQ ID	LLSSRVGYMH	DTMYQAS	FQGSGYPFT
	NO:316	NO:317	(SEQ ID	PSLKD	(SEQ ID	NO:318	NO:319	(SEQ ID	(SEQ ID	(SEQ ID
			NO:10)	(SEQ ID NO:19)	NO:29)			NO:320)	NO:321)	NO:6)
***A17b5	SEQ ID	SEQ ID	TAGMSVG	DIWWDDKKHYN	DMIFNWYFDV	SEQ ID	SEQ ID	SLSSRVGYMH	DTYYLPS	FQGSGYPFT
	NO:322	NO:323	(SEQ ID	PSLKD	(SEQ ID	NO:324	NO:325	(SEQ ID	(SEQ ID	(SEQ ID
			NO:10)	(SEQ ID NO:19)	NO:29)			NO:22)	NO:326)	NO:6)
***A17f5	SEQ ID	SEQ ID	TAGMSVG	DIWWDDKKDYN	DMIFNWYFDV	SEQ ID	SEQ ID	SLSSRVGYMH	DTFRHTS	FQGSGYPFT
	NO:327	NO:328	(SEQ ID	PSLKD	(SEQ ID	NO:330	NO:332	(SEQ ID	(SEQ ID	(SEQ ID
			NO:10	(SEQ ID NO:	NO:29)			NO:22)	NO:332)	NO:6)
				329)
***A17h4	SEQ ID	SEQ ID	TAGMSVG	DIWWDGKKHYN	DMIFNWYFDV	SEQ ID	SEQ ID	SPSSSVGYMH	DTYYLAS	FQGSGYPFT
	NO:218	NO:33	(SEQ ID	PSLKD	(SEQ ID	NO:333	NO:334	(SEQ ID	(SEQ ID	(SEQ ID
			NO:10)	(SEQ ID NO:25)	NO:29)			NO:335)	NO:336)	NO:6)
Bold faced & underlined amino acid residues are the residues which differ from the amino acid sequence in palivizumab;
Fab fragment produced*;
Monoclonal antibody produced**;
Fab fragment & monoclonal antibody produced***

[0183]

TABLE 3A
VH CDR1 Sequences
	SVG	(SEQ ID NO:1)
	SVG	(SEQ ID NO:10)
	SVG	(SEQ ID NO:18)
	Bold faced & underlined amino acid residues are the residues which differ from the amino acid sequence in palivizumab

[0184]

TABLE 3B
VH CDR2 Sequences
DDKKDYNPSLKS	(SEQ ID NO:2)	DGKKDYNPSLKS	(SEQ ID NO:100)
DDKKDYNPSLKD	(SEQ ID NO:86)	DGKKDYNPSLKD	(SEQ ID NO:103)
DDKKHYNPSLKS	(SEQ ID NO:82)	DGKKHYNPSLKS	(SEQ ID NO:106)
DDKKHYNPSLKD	(SEQ ID NO:19)	DGKKHYNPSLKD	(SEQ ID NO:25)
DDKKSYNPSLKS	(SEQ ID NO:109)	DGKKSYNPSLKS	(SEQ ID NO:114)
DDKKSYNPSLKD	(SEQ ID NO:111)	DGKKSYNPSLKD	(SEQ ID NO:41)
DDKGDYNPSLKS	(SEQ ID NO:384)	DGKGDYNPSLKS	(SEQ ID NO:390)
DDKGDYNPSLKD	(SEQ ID NO:385)	DGKGDYNPSLKD	(SEQ ID NO:391)
DDKGHYNPSLKS	(SEQ ID NO:386)	DGKGHYNPSLKS	(SEQ ID NO:392)
DDKGHYNPSLKD	(SEQ ID NO:387)	DGKGHYNPSLKD	(SEQ ID NO:393)
DDKGSYNPSLKS	(SEQ ID NO:388)	DGKGSYNPSLKS	(SEQ ID NO:394)
DDKGSYNPSLKD	(SEQ ID NO:389)	DGKGSYNPSLKD	(SEQ ID NO:395)
Bold faced & underlined amino acid residues are the residues which differ from the amino acid sequence in palivizumab

[0185]

TABLE 3C
VH CDR3 Sequences
	NWYFDV	(SEQ ID NO:3)	NWYFDV	(SEQ ID NO:83)
	NFYFDV	(SEQ ID NO:12)	NFYFDV	(SEQ ID NO:29)
	NWYFDV	(SEQ ID NO:94)	NWYFDV	(SEQ ID NO:79)
	NFYFDV	(SEQ ID NO:97)	NFYFDV	(SEQ ID NO:20)
	Bold faced & underlined amino acid residues are the residues which differ from the amino acid sequence in palivizumab

[0186]

TABLE 3D
VL CDR1 Sequences
KCQLSVGYMH	(SEQ ID NO:4)	SCQLSVGYMH	(SEQ ID NO:127)	LCQLSVGYMH	(SEQ ID NO:204)
RVGYMH	(SEQ ID NO:87)	RVGYMH	(SEQ ID NO:132)	RVGYMH	(SEQ ID NO:206)
KCQLFVGYMH	(SEQ ID NO:396)	SCQLFVGYMH	(SEQ ID NO:436)	LCQLFVGYMH	(SEQ ID NO:476)
KCQSSVGYMH	(SEQ ID NO:80)	SCQSSVGYMH	(SEQ ID NO:129)	LCQSSVGYMH	(SEQ ID NO:205)
KCQSRVGYMH	(SEQ ID NO:84)	SCQSRVGYMH	(SEQ ID NO:130)	LCQSRVGYMH	(SEQ ID NO:203)
KCQSFVGYMH	(SEQ ID NO:397)	SCQSFVGYMH	(SEQ ID NO:437)	LCQSFVGYMH	(SEQ ID NO:477)
KCQVSVGYMH	(SEQ ID NO:398)	SCQVSVGYMH	(SEQ ID NO:438)	LCQVSVGYMH	(SEQ ID NO:478)
KCQVRVGYMH	(SEQ ID NO:399)	SCQVRVGYMH	(SEQ ID NO:439)	LCQVRVGYMH	(SEQ ID NO:479)
KCQVFVGYMH	(SEQ ID NO:400)	SCQVFVGYMH	(SEQ ID NO:440)	LCQVFVGYMH	(SEQ ID NO:480)
KCSLSVGYMH	(SEQ ID NO:112)	SCSLSVGYMH	(SEQ ID NO:142)	LCSLSVGYMH	(SEQ ID NO:196)
KCSLRVGYMH	(SEQ ID NO:119)	SCSLRVGYMH	(SEQ ID NO:148)	LCSLRVGYMH	(SEQ ID NO:198)
KCSLFVGYMH	(SEQ ID NO:401)	SCSLFVGYMH	(SEQ ID NO:441)	LCSLFVGYMH	(SEQ ID NO:481)
KCSSSVGYMH	(SEQ ID NO:115)	SCSSSVGYMH	(SEQ ID NO:144)	LCSSSVGYMH	(SEQ ID NO:197)
KCSSRVGYMH	(SEQ ID NO:117)	SCSSRVGYMH	(SEQ ID NO:146)	LCSSRVGYMH	(SEQ ID NO:195)
KCSSFVGYMH	(SEQ ID NO:402)	SCSSFVGYMH	(SEQ ID NO:442)	LCSSFVGYMH	(SEQ ID NO:482)
KCSVSVGYMH	(SEQ ID NO:403)	SCSVSVGYMH	(SEQ ID NO:443)	LCSVSVGYMH	(SEQ ID NO:483)
KCSVRVGYMH	(SEQ ID NO:404)	SCSVRVGYMH	(SEQ ID NO:444)	LCSVRVGYMH	(SEQ ID NO:484)
KCSVFVGYMH	(SEQ ID NO:405)	SCSVFVGYMH	(SEQ ID NO:445)	LCSVFVGYMH	(SEQ ID NO:485)
KAQLSVGYMH	(SEQ ID NO:182)	SAQLSVGYMH	(SEQ ID NO:207)	LAQLSVGYMH	(SEQ ID NO:486)
KAQLRVGYMH	(SEQ ID NO:180)	SAQLRVGYMH	(SEQ ID NO:190)	LAQLRVGYMH	(SEQ ID NO:487)
KAQLFVGYMH	(SEQ ID NO:406)	SAQLFVGYMH	(SEQ ID NO:446)	LAQLFVGYMH	(SEQ ID NO:488)
KAQSSVGYMH	(SEQ ID NO:181)	SAQSSVGYMH	(SEQ ID NO:191)	LAQSSVGYMH	(SEQ ID NO:489)
KAQSRVGYMH	(SEQ ID NO:179)	SAQSRVGYMH	(SEQ ID NO:189)	LAQSRVGYMH	(SEQ ID NO:490)
KAQSFVGYMH	(SEQ ID NO:407)	SAQSFVGYMH	(SEQ ID NO:447)	LAQSFVGYMH	(SEQ ID NO:491)
KAQVSVGYMH	(SEQ ID NO:408)	SAQVSVGYMH	(SEQ ID NO:448)	LAQVSVGYMH	(SEQ ID NO:492)
KAQVRVGYMH	(SEQ ID NO:409)	SAQVRVGYMH	(SEQ ID NO:449)	LAQVRVGYMH	(SEQ ID NO:493)
KAQVFVGYMH	(SEQ ID NO:410)	SAQVFVGYMH	(SEQ ID NO:450)	LAQVFVGYMH	(SEQ ID NO:494)
KASLSVGYMH	(SEQ ID NO:186)	SASLSVGYMH	(SEQ ID NO:188)	LASLSVGYMH	(SEQ ID NO:495)
KASLRVGYMH	(SEQ ID NO:184)	SASLRVGYMH	(SEQ ID NO:187)	LASLRVGYMH	(SEQ ID NO:496)
KASLFVGYMH	(SEQ ID NO:411)	SASLFVGYMH	(SEQ ID NO:451)	LASLFVGYMH	(SEQ ID NO:497)
KASSSVGYMH	(SEQ ID NO:185)	SASSSVGYMH	(SEQ ID NO:14)	LASSSVGYMH	(SEQ ID NO:498)
KASSRVGYMH	(SEQ ID NO:183)	SASSRVGYMH	(SEQ ID NO:39)	LASSRVGYMH	(SEQ ID NO:499)
KASSFVGYMH	(SEQ ID NO:412)	SASSFVGYMH	(SEQ ID NO:452)	LASSFVGYMH	(SEQ ID NO:500)
KASVSVGYMH	(SEQ ID NO:413)	SASVSVGYMH	(SEQ ID NO:453)	LASVSVGYMH	(SEQ ID NO:501)
KASVRVGYMH	(SEQ ID NO:414)	SASVRVGYMH	(SEQ ID NO:454)	LASVRVGYMH	(SEQ ID NO:502)
KASVFVGYMH	(SEQ ID NO:415)	SASVFVGYMH	(SEQ ID NO:455)	LASVFVGYMH	(SEQ ID NO:503)
KLQLSVGYMH	(SEQ ID NO:89)	SLQLSVGYMH	(SEQ ID NO:134)	LLQLSVGYMH	(SEQ ID NO:504)
KLQLRVGYMH	(SEQ ID NO:98)	SLQLRVGYMH	(SEQ ID NO:140)	LLQLRVGYMH	(SEQ ID NO:505)
KLQLFVGYMH	(SEQ ID NO:416)	SLQLFVGYMH	(SEQ ID NO:456)	LLQLFVGYMH	(SEQ ID NO:506)
KLQSSVGYMH	(SEQ ID NO:92)	SLQSSVGYMH	(SEQ ID NO:136)	LLQSSVGYMH	(SEQ ID NO:507)
KLQSRVGYMH	(SEQ ID NO:95)	SLQSRVGYMH	(SEQ ID NO:138)	LLQSRVGYMH	(SEQ ID NO:508)
KLQSFVGYMH	(SEQ ID NO:417)	SLQSFVGYMH	(SEQ ID NO:457)	LLQSFVGYMH	(SEQ ID NO:509)
KLQVSVGYMH	(SEQ ID NO:418)	SLQVSVGYMH	(SEQ ID NO:458)	LLQVSVGYMH	(SEQ ID NO:510)
KLQVRVGYMH	(SEQ ID NO:419)	SLQVRVGYMH	(SEQ ID NO:459)	LLQVRVGYMH	(SEQ ID NO:511)
KLQVFVGYMH	(SEQ ID NO:420)	SLQVFVGYMH	(SEQ ID NO:460)	LLQVFVGYMH	(SEQ ID NO:512)
KLSLSVGYMH	(SEQ ID NO:101)	SLSLSVGYMH	(SEQ ID NO:120)	LLSLSVGYMH	(SEQ ID NO:513)
KLSLRVGYMH	(SEQ ID NO:110)	SLSLRVGYMH	(SEQ ID NO:125)	LLSLRVGYMH	(SEQ ID NO:514)
KLSLFVGYMH	(SEQ ID NO:421)	SLSLFVGYMH	(SEQ ID NO:461)	LLSLFVGYMH	(SEQ ID NO:515)
KLSSSVGYMH	(SEQ ID NO:104)	SLSSSVGYMH	(SEQ ID NO:122)	LLSSSVGYMH	(SEQ ID NO:516)
KLSSRVGYMH	(SEQ ID NO:107)	SLSSRVGYMH	(SEQ ID NO:22)	LLSSRVGYMH	(SEQ ID NO:517)
KLSSFVGYMH	(SEQ ID NO:422)	SLSSFVGYMH	(SEQ ID NO:462)	LLSSFVGYMH	(SEQ ID NO:518)
KLSVSVGYMH	(SEQ ID NO:423)	SLSVSVGYMH	(SEQ ID NO:463)	LLSVSVGYMH	(SEQ ID NO:519)
KLSVRVGYMH	(SEQ ID NO:424)	SLSVRVGYMH	(SEQ ID NO:464)	LLSVRVGYMH	(SEQ ID NO:520)
KLSVFVGYMH	(SEQ ID NO:425)	SLSVFVGYMH	(SEQ ID NO:465)	LLSVFVGYMH	(SEQ ID NO:521)
KPQLSVGYMH	(SEQ ID NO:163)	SPQLSVGYMH	(SEQ ID NO:177)	LPQLSVGYMH	(SEQ ID NO:200)
KPQLRVGYMH	(SEQ ID NO:159)	SPQLRVGYMH	(SEQ ID NO:173)	LPQLRVGYMH	(SEQ ID NO:202)
KPQLFVGYMH	(SEQ ID NO:426)	SPQLFVGYMH	(SEQ ID NO:466)	LPQLFVGYMH	(SEQ ID NO:522)
KPQSSVGYMH	(SEQ ID NO:161)	SPQSSVGYMH	(SEQ ID NO:176)	LPQSSVGYMH	(SEQ ID NO:201)
KPQSRVGYMH	(SEQ ID NO:157)	SPQSRVGYMH	(SEQ ID NO:171)	LPQSRVGYMH	(SEQ ID NO:199)
KPQSFVGYMH	(SEQ ID NO:427)	SPQSFVGYMH	(SEQ ID NO:467)	LPQSFVGYMH	(SEQ ID NO:523)
KPQVSVGYMH	(SEQ ID NO:428)	SPQVSVGYMH	(SEQ ID NO:468)	LPQVSVGYMH	(SEQ ID NO:524)
KPQVRVGYMH	(SEQ ID NO:429)	SPQVRVGYMH	(SEQ ID NO:469)	LPQVRVGYMH	(SEQ ID NO:525)
KPQVFVGYMH	(SEQ ID NO:430)	SPQVFVGYMH	(SEQ ID NO:470)	LPQVFVGYMH	(SEQ ID NO:526)
KPSLSVGYMH	(SEQ ID NO:155)	SPSLSVGYMH	(SEQ ID NO:169)	LPSLSVGYMH	(SEQ ID NO:192)
KPSLRVGYMH	(SEQ ID NO:152)	SPSee LRVGYMH	(SEQ ID NO:166)	LPSLRVGYMH	(SEQ ID NO:194)
KPSLFVGYMH	(SEQ ID NO:431)	SPSLFVGYMH	(SEQ ID NO:471)	LPSLFVGYMH	(SEQ ID NO:527)
KPSSSVGYMH	(SEQ ID NO:153)	SPSSSVGYMH	(SEQ ID NO:168)	LPSSSVGYMH	(SEQ ID NO:193)
KPSSRVGYMH	(SEQ ID NO:150)	SPSSRVGYMH	(SEQ ID NO:31)	LPSSRVGYMH	(SEQ ID NO:47)
KPSSFVGYMH	(SEQ ID NO:432)	SPSSFVGYMH	(SEQ ID NO:472)	LPSSFVGYMH	(SEQ ID NO:528)
KPSVSVGYMH	(SEQ ID NO:433)	SPSVSVGYMH	(SEQ ID NO:473)	LPSVSVGYMH	(SEQ ID NO:529)
KPSVRVGYMH	(SEQ ID NO:434)	SPSVRVGYMH	(SEQ ID NO:474)	LPSVRVGYMH	(SEQ ID NO:530)
KPSVFVGYMH	(SEQ ID NO:435)	SPSVFVGYMH	(SEQ ID NO:475)	LPSVFVGYMH	(SEQ ID NO:531)
Bold faced & underlined amino acid residues are the residues which differ from the amino acid sequence in palivizumab

[0187]

TABLE 3E
VL CDR2 Sequences
DTSKLAS (SEQ	DTFKLAS (SEQ	DTYKLAS (SEQ	DTRKLAS (SEQ	DTMKLAS (SEQ	DTKKLAS (SEQ	DTLKLAS (SEQ
ID NO:5)	ID NO:15)	ID NO:799)	ID NOS:	ID NOS:	ID NO:1211)	ID NO:135)
			113&174)	121&162)
DTSKLSS (SEQ	DTFKLSS (SEQ	DTYKLSS (SEQ	DTRKLSS (SEQ	DTMKLSS (SEQ	DTKKLSS (SEQ	DTLKLSS (SEQ
ID NO:165)	ID NO:96)	ID NO:800)	ID NO:175)	ID NO:164)	ID NO:1212)	ID NO:1355)
DTSKLKS (SEQ	DTFKLKS (SEQ	DTYKLKS (SEQ	DTRKLKS (SEQ	DTMKLKS (SEQ	DTKKLKS (SEQ	DTLKLKS (SEQ
ID NO:532)	ID NO:660)	ID NO:801)	ID NO:943)	ID NO:1076)	ID NO:1213)	ID NO:1356)
DTSKLRS (SEQ	DTFKLRS (SEQ	DTYKLRS (SEQ	DTRKLRS (SEQ	DTMKLRS (SEQ	DTKKLRS (SEQ	DTLKLRS (SEQ
ID NO:533)	ID NO:661)	ID NO:802)	ID NO:944)	ID NO:1077)	ID NO:1214)	ID NO:1357)
DTSKLHS (SEQ	DTFKLHS (SEQ	DTYKLHS (SEQ	DTRKLHS (SEQ	DTMKLHS (SEQ	DTKKLHS (SEQ	DTLKLHS (SEQ
ID NO:534)	ID NO:662)	ID NO:803)	ID NO:945)	ID NO:1078)	ID NO:1215)	ID NO:1358)
DTSKLPS (SEQ	DTFKLPS (SEQ	DTYKLPS (SEQ	DTRKLPS (SEQ	DTMKLPS (SEQ	DTKKLPS (SEQ	DTLKLPS (SEQ
ID NO:102)	ID NO:663)	ID NO:804)	ID NO:118)	ID NO:1079)	ID NO:1216)	ID NO:1359)
DTSKLTS (SEQ	DTFKLTS (SEQ	DTYKLTS (SEQ	DTRKLTS (SEQ	DTMKLTS (SEQ	DTKKLTS (SEQ	DTLKLTS (SEQ
ID NO:535)	ID NO:664)	ID NO:805)	ID NO:946)	ID NO:1080)	ID NO:1217)	ID NO:1360)
DTSKLDS (SEQ	DTFKLDS (SEQ	DTYKLDS (SEQ	DTRKLDS (SEQ	DTMKLDS (SEQ	DTKKLDS (SEQ	DTLKLDS(SEQ
ID NO:128)	ID NO:665)	ID NO:806)	ID NO:947)	ID NO:1081)	ID NO:1218)	ID NO:131)
DTSKHAS (SEQ	DTFKHAS (SEQ	DTYKHAS (SEQ	DTRKHAS (SEQ	DTMKHAS (SEQ	DTKKHAS (SEQ	DTLKHAS (SEQ
ID NO:536)	ID NO:666)	ID NO:807)	ID NO:948)	ID NO:1082)	ID NO:1219)	ID NO:1361)
DTSKHSS (SEQ	DTFKHSS (SEQ	DTYKHSS (SEQ	DTRKHSS (SEQ	DTMKHSS (SEQ	DTKKHSS (SEQ	DTLKHSS (SEQ
ID NO:537)	ID NO:667)	ID NO:808)	ID NO:949)	ID NO:1083)	ID NO:1220)	ID NO:1362)
DTSKHKS (SEQ	DTFKHKS (SEQ	DTYKHKS (SEQ	DTRKHKS (SEQ	DTMKHKS (SEQ	DTKKHKS (SEQ	DTLKHKS (SEQ
ID NO:538)	ID NO:668)	ID NO:809)	ID NO:950)	ID NO:1084)	ID NO:1221)	ID NO:1363)
DTSKHRS (SEQ	DTFKHRS(SEQ	DTYKHRS (SEQ	DTRKHRS (SEQ	DTMKHRS (SEQ	DTKKHRS (SEQ	DTLKHRS (SEQ
ID NO:539)	ID NO:669)	ID NO:810)	ID NO:951)	ID NO:1085)	ID NO:1222)	ID NO:1364)
DTSKHHS (SEQ	DTFKHHS (SEQ	DTYKHHS (SEQ	DTRKHHS (SEQ	DTMKHHS (SEQ	DTKKHHS (SEQ	DTLKHHS (SEQ
ID NO:540)	ID NO:670)	ID NO:811)	ID NO:952)	ID NO:1086)	ID NO:1223)	ID NO:1365)
DTSKHPS (SEQ	DTFKHPS (SEQ	DTYKHPS (SEQ	DTRKHPS (SEQ	DTMKHPS (SEQ	DTKKHPS (SEQ	DTLKHPS (SEQ
ID NO:541)	ID NO:671)	ID NO:812)	ID NO:953)	ID NO:1087)	ID NO:1224)	ID NO:1366)
DTSKHTS (SEQ	DTFKHTS (SEQ	DTYKHTS (SEQ	DTRKHTS (SEQ	DTMKHTS (SEQ	DTKKHTS (SEQ	DTLKHTS (SEQ
ID NO:542)	ID NO:672)	ID NO:813)	ID NO:954)	ID NO:1088)	ID NO:1225)	ID NO:1367)
DTSKHDS (SEQ	DTFKHDS (SEQ	DTYKHDS (SEQ	DTRKHDS (SEQ	DTMKHDS (SEQ	DTKKHDS (SEQ	DTLKHDS (SEQ
ID NO:543)	ID NO:673)	ID NO:814)	ID NO:955)	ID NO:1089)	ID NO:1226)	ID NO:1368)
DTSKQAS (SEQ	DTFKQAS (SEQ	DTYKQAS (SEQ	DTRKQAS (SEQ	DTMKQAS (SEQ	DTKKQAS (SEQ	DTLKQAS (SEQ
ID NO:139)	ID NO:674)	ID NO:815)	ID NO:170)	ID NO:154)	ID NO:1227)	ID NO:1369)
DTSKQSS (SEQ	DTFKQSS (SEQ	DTYKQSS (SEQ	DTRKQSS (SEQ	DTMKQSS (SEQ	DTKKQSS (SEQ	DTLKQSS (SEQ
ID NO:141)	ID NO:675)	ID NO:816)	ID NO:172)	ID NO:156)	ID NO:1228)	ID NO:1370)
DTSKQKS (SEQ	DTFKQKS (SEQ	DTYKQKS (SEQ	DTRKQKS (SEQ	DTMKQKS (SEQ	DTKKQKS (SEQ	DTLKQKS (SEQ
ID NO:544)	ID NO:676)	ID NO:817)	ID NO:956)	ID NO:1090)	ID NO:1229)	ID NO:1371)
DTSKQRS (SEQ	DTFKQRS (SEQ	DTYKQRS (SEQ	DTRKQRS (SEQ	DTMKQRS (SEQ	DTKKQRS (SEQ	DTLKQRS (SEQ
ID NO:545)	ID NO:677)	ID NO:818)	ID NO:957)	ID NO:1091)	ID NO:1230)	ID NO:1372)
DTSKQHS (SEQ	DTFKQHS (SEQ	DTYKQHS (SEQ	DTRKQHS (SEQ	DTMKQHS (SEQ	DTKKQHS (SEQ	DTLKQHS (SEQ
ID NO:546)	ID NO:678)	ID NO:819)	ID NO:958)	ID NO:1092)	ID NO:1231)	ID NO:1373)
DTSKQPS (SEQ	DTFKQPS (SEQ	DTYKQPS (SEQ	DTRKQPS (SEQ	DTMKQPS (SEQ	DTKKQPS (SEQ	DTLKQPS (SEQ
ID NO:547)	ID NO:679)	ID NO:820)	ID NO:959)	ID NO:1093)	ID NO:1232)	ID NO:1374)
DTSKQTS (SEQ	DTFKQTS (SEQ	DTYKQTS (SEQ	DTRKQTS (SEQ	DTMKQTS (SEQ	DTKKQTS (SEQ	DTLKQTS (SEQ
ID NO:548)	ID NO:680)	ID NO:821)	ID NO:960)	ID NO:1094)	ID NO:1233)	ID NO:1375)
DTSKQDS (SEQ	DTFKQDS (SEQ	DTYKQDS (SEQ	DTRKQDS (SEQ	DTMKQDS (SEQ	DTKKQDS (SEQ	DTLKQDS (SEQ
ID NO:549)	ID NO:681)	ID NO:822)	ID NO:961)	ID NO:1095)	ID NO:1234)	ID NO:1376)
DTSGLAS (SEQ	DTFGLAS (SEQ	DTYGLAS (SEQ	DTRGLAS (SEQ	DTMGLAS (SEQ	DTKGLAS (SEQ	DTLGLAS (SEQ
ID NO:105)	ID NO:682)	ID NO:823)	ID NO:116)	ID NO:1096)	ID NO:1235)	ID NO:1377)
DTSGLSS (SEQ	DTFGLSS (SEQ	DTYGLSS (SEQ	DTRGLSS (SEQ	DTMGLSS (SEQ	DTKGLSS (SEQ	DTLGLSS (SEQ
ID NO:550)	ID NO:683)	ID NO:824)	ID NO:962)	ID NO:1097)	ID NO:1236)	ID NO:1378)
DTSGLKS (SEQ	DTFGLKS (SEQ	DTYGLKS (SEQ	DTRGLKS (SEQ	DTMGLKS (SEQ	DTKGLKS (SEQ	DTLGLKS (SEQ
ID NO:551)	ID NO:684)	ID NO:825)	ID NO:963)	ID NO:1098)	ID NO:1237)	ID NO:1379)
DTSGLRS (SEQ	DTFGLRS (SEQ	DTYGLRS (SEQ	DTRGLRS (SEQ	DTMGLRS (SEQ	DTKGLRS (SEQ	DTLGLRS (SEQ
ID NO:552)	ID NO:685)	ID NO:826)	ID NO:964)	ID NO:1099)	ID NO:1238)	ID NO:1380)
DTSGLHS (SEQ	DTFGLHS (SEQ	DTYGLHS (SEQ	DTRGLHS (SEQ	DTMGLHS (SEQ	DTKGLHS (SEQ	DTLGLHS (SEQ
ID NO:553)	ID NO:686)	ID NO:827)	ID NO:965)	ID NO:1100)	ID NO:1239)	ID NO:1381)
DTSGLPS (SEQ	DTFGLPS (SEQ	DTYGLPS (SEQ	DTRGLPS (SEQ	DTMGLPS (SEQ	DTKGLPS (SEQ	DTLGLPS (SEQ
ID NO:108)	ID NO:687)	ID NO:828)	ID NO:27)	ID NO:1101)	ID NO:1240)	ID NO:1382)
DTSGLTS (SEQ	DTFGLTS (SEQ	DTYGLTS (SEQ	DTRGLTS (SEQ	DTMGLTS (SEQ	DTKGLTS (SEQ	DTLGLTS (SEQ
ID NO:554)	ID NO:688)	ID NO:829)	ID NO:966)	ID NO:1102)	ID NO:1241)	ID NO:1383)
DTSGLDS (SEQ	DTFGLDS (SEQ	DTYGLDS (SEQ	DTRGLDS (SEQ	DTMGLDS (SEQ	DTKGLDS (SEQ	DTLGLDS (SEQ
ID NO:555)	ID NO:689)	ID NO:830)	ID NO:967)	ID NO:1103)	ID NO:1242)	ID NO:1384)
DTSGHAS (SEQ	DTFGHAS (SEQ	DTYGHAS (SEQ	DTRGHAS (SEQ	DTMGHAS (SEQ	DTKGHAS (SEQ	DTLGHAS (SEQ
ID NO:556)	ID NO:690)	ID NO:831)	ID NO:968)	ID NO:1104)	ID NO:1243)	ID NO:1385)
DTSGHSS (SEQ	DTFGHSS (SEQ	DTYGHSS (SEQ	DTRGHSS (SEQ	DTMGHSS (SEQ	DTKGHSS (SEQ	DTLGHSS (SEQ
ID NO:557)	ID NO:691)	ID NO:832)	ID NO:969)	ID NO:1105)	ID NO:1244)	ID NO:1386)
DTSGHKS (SEQ	DTFGHKS (SEQ	DTYGHKS (SEQ	DTRGHKS (SEQ	DTMGHKS (SEQ	DTKGHKS (SEQ	DTLGHKS (SEQ
ID NO:558)	ID NO:692)	ID NO:833)	ID NO:970)	ID NO:1106)	ID NO:1245)	ID NO:1387)
DTSGHRS (SEQ	DTFGHRS (SEQ	DTYGHRS (SEQ	DTRGHRS (SEQ	DTMGHRS (SEQ	DTKGHRS (SEQ	DTLGHRS (SEQ
ID NO:559)	ID NO:693)	ID NO:834)	ID NO:971)	ID NO:1107)	ID NO:1246)	ID NO:1388)
DTSGHHS (SEQ	DTFGHHS (SEQ	DTYGHHS (SEQ	DTRGHHS (SEQ	DTMGHHS (SEQ	DTKGHHS (SEQ	DTLGHHS (SEQ
ID NO:560)	ID NO:694)	ID NO:835)	ID NO:972)	ID NO:1108)	ID NO:1247)	ID NO:1389)
DTSGHPS (SEQ	DTFGHPS (SEQ	DTYGHPS (SEQ	DTRGHPS (SEQ	DTMGHPS (SEQ	DTKGHPS (SEQ	DTLGHPS (SEQ
ID NO:561)	ID NO:695)	ID NO:836)	ID NO:973)	ID NO:1109)	ID NO:1248)	ID NO:1390)
DTSGHTS (SEQ	DTFGHTS (SEQ	DTYGHTS (SEQ	DTRGHTS (SEQ	DTMGHTS (SEQ	DTKGHTS (SEQ	DTLGHTS (SEQ
ID NO:562)	ID NO:696)	ID NO:837)	ID NO:974)	ID NO:1110)	ID NO:1249)	ID NO:1391)
DTSGHDS (SEQ	DTFGHDS (SEQ	DTYGHDS (SEQ	DTRGHDS (SEQ	DTMGHDS (SEQ	DTKGHDS (SEQ	DTLGHDS (SEQ
ID NO:563)	ID NO:697)	ID NO:838)	ID NO:975)	ID NO:1111)	ID NO:1250)	ID NO:1392)
DTSGQAS (SEQ	DTFGQAS (SEQ	DTYGQAS (SEQ	DTRGQAS (SEQ	DTMGQAS (SEQ	DTKGQAS (SEQ	DTLGQAS (SEQ
ID NO:564)	ID NO:698)	ID NO:839)	ID NO:976)	ID NO:1112)	ID NO:1251)	ID NO:1393)
DTSGQSS (SEQ	DTFGQSS (SEQ	DTYGQSS (SEQ	DTRGQSS (SEQ	DTMGQSS (SEQ	DTKGQSS (SEQ	DTLGQSS (SEQ
ID NO:565)	ID NO:699)	ID NO:840)	ID NO:977)	ID NO:1113)	ID NO:1252)	ID NO:1394)
DTSGQKS (SEQ	DTFGQKS (SEQ	DTYGQKS (SEQ	DTRGQKS (SEQ	DTMGQKS (SEQ	DTKGQKS (SEQ	DTLGQKS (SEQ
ID NO:566)	ID NO:700)	ID NO:841)	ID NO:978)	ID NO:1114)	ID NO:1253)	ID NO:1395)
DTSGQRS (SEQ	DTFGQRS (SEQ	DTYGQRS (SEQ	DTRGQRS (SEQ	DTMGQRS (SEQ	DTKGQRS (SEQ	DTLGQRS (SEQ
ID NO:567)	ID NO:701)	ID NO:842)	ID NO:979)	ID NO:1115)	ID NO:1254)	ID NO:1396)
DTSGQHS (SEQ	DTFGQHS (SEQ	DTYGQHS (SEQ	DTRGQHS (SEQ	DTMGQHS (SEQ	DTKGQHS (SEQ	DTLGQHS (SEQ
ID NO:568)	ID NO:702)	ID NO:843)	ID NO:980)	ID NO:1116)	ID NO:1255)	ID NO:1397)
DTSGQPS (SEQ	DTFGQPS (SEQ	DTYGQPS (SEQ	DTRGQPS (SEQ	DTMGQPS (SEQ	DTKGQPS (SEQ	DTLGQPS (SEQ
ID NO:569)	ID NO:703)	ID NO:844)	ID NO:981)	ID NO:1117)	ID NO:1256)	ID NO:1398)
DTSGQTS (SEQ	DTFGQTS (SEQ	DTYGQTS (SEQ	DTRGQTS (SEQ	DTMGQTS (SEQ	DTKGQTS (SEQ	DTLGQTS (SEQ
ID NO:570)	ID NO:704)	ID NO:845)	ID NO:982)	ID NO:1118)	ID NO:1257)	ID NO:1399)
DTSGQDS (SEQ	DTFGQDS (SEQ	DTYGQDS (SEQ	DTRGQDS (SEQ	DTMGQDS (SEQ	DTKGQDS (SEQ	DTLGQDS (SEQ
ID NO:571)	ID NO:705)	ID NO:846)	ID NO:983)	ID NO:1119)	ID NO:1258)	ID NO:1400)
DTSRLAS (SEQ	DTFRLAS (SEQ	DTYRLAS (SEQ	DTRRLAS (SEQ	DTMRLAS (SEQ	DTKRLAS (SEQ	DTLRLAS (SEQ
ID NO:123)	ID NO:706)	ID NO:847)	ID NO:984)	ID NO:32)	ID NO:1259)	ID NO:1401)
DTSRLSS (SEQ	DTFRLSS (SEQ	DTYRLSS (SEQ	DTRRLSS (SEQ	DTMRLSS (SEQ	DTKRLSS (SEQ	DTLRLSS (SEQ
ID NO:572)	ID NO:707)	ID NO:848)	ID NO:985)	ID NO:1120)	ID NO:1260)	ID NO:1402)
DTSRLKS (SEQ	DTFRLKS (SEQ	DTYRLKS (SEQ	DTRRLKS (SEQ	DTMRLKS (SEQ	DTKRLKS (SEQ	DTLRLKS (SEQ
ID NO:573)	ID NO:708)	ID NO:849)	ID NO:986)	ID NO:1121)	ID NO:1261)	ID NO:1403)
DTSRLRS (SEQ	DTFRLRS (SEQ	DTYRLRS (SEQ	DTRRLRS (SEQ	DTMRLRS (SEQ	DTKRLRS (SEQ	DTLRLRS (SEQ
ID NO:574)	ID NO:709)	ID NO:850)	ID NO:987)	ID NO:1122)	ID NO:1262)	ID NO:1404)
DTSRLHS (SEQ	DTFRLHS (SEQ	DTYRLHS (SEQ	DTRRLHS (SEQ	DTMRLHS (SEQ	DTKRLHS (SEQ	DTLRLHS (SEQ
ID NO:575)	ID NO:710)	ID NO:851)	ID NO:988)	ID NO:1123)	ID NO:1263)	ID NO:1405)
DTSRLPS (SEQ	DTFRLPS (SEQ	DTYRLPS (SEQ	DTRRLPS (SEQ	DTMRLPS (SEQ	DTKRLPS (SEQ	DTLRLPS (SEQ
ID NO:576)	ID NO:711)	ID NO:852)	ID NO:989)	ID NO:1124)	ID NO:1264)	ID NO:1406)
DTSRLTS (SEQ	DTFRLTS (SEQ	DTYRLTS (SEQ	DTRRLTS (SEQ	DTMRLTS (SEQ	DTKRLTS (SEQ	DTLRLTS (SEQ
ID NO:577)	ID NO:712)	ID NO:853)	ID NO:990)	ID NO:1125)	ID NO:1265)	ID NO:1407)
DTSRLDS (SEQ	DTFRLDS (SEQ	DTYRLDS (SEQ	DTRRLDS (SEQ	DTMRLDS (SEQ	DTKRLDS (SEQ	DTLRLDS (SEQ
ID NO:578)	ID NO:713)	ID NO:854)	ID NO:991)	ID NO:1126)	ID NO:1266)	ID NO:1408)
DTSRHAS (SEQ	DTFRHAS (SEQ	DTYRHAS (SEQ	DTRRHAS (SEQ	DTMRHAS (SEQ	DTKRHAS (SEQ	DTLRHAS (SEQ
ID NO:579)	ID NO:714)	ID NO:855)	ID NO:992)	ID NO:1127)	ID NO:1267)	ID NO:1409)
DTSRHSS (SEQ	DTFRHSS (SEQ	DTYRHSS (SEQ	DTRRHSS (SEQ	DTMRHSS (SEQ	DTKRHSS (SEQ	DTLRHSS (SEQ
ID NO:580)	ID NO:715)	ID NO:856)	ID NO:993)	ID NO:1128)	ID NO:1268)	ID NO:1410)
DTSRHKS (SEQ	DTFRHKS (SEQ	DTYRHKS (SEQ	DTRRHKS (SEQ	DTMRHKS (SEQ	DTKRHKS (SEQ	DTLRHKS (SEQ
ID NO:581)	ID NO:716)	ID NO:857)	ID NO:994)	ID NO:1129)	ID NO:1269)	ID NO:1411)
DTSRHRS (SEQ	DTFRHRS (SEQ	DTYRHRS (SEQ	DTRRHRS (SEQ	DTMRHRS (SEQ	DTKRHRS (SEQ	DTLRHRS (SEQ
ID NO:582)	ID NO:717)	ID NO:858)	ID NO:995)	ID NO:1130)	ID NO:1270)	ID NO:1412)
DTSRHHS (SEQ	DTFRHHS (SEQ	DTYRHHS (SEQ	DTRRHHS (SEQ	DTMRHHS (SEQ	DTKRHHS (SEQ	DTLRHHS (SEQ
ID NO:583)	ID NO:718)	ID NO:859)	ID NO:996)	ID NO:1131)	ID NO:1271)	ID NO:1413)
DTSRHPS (SEQ	DTFRHPS (SEQ	DTYRHPS (SEQ	DTRRHPS (SEQ	DTMRHPS (SEQ	DTKRHPS (SEQ	DTLRHPS (SEQ
ID NO:584)	ID NO:719)	ID NO:860)	ID NO:997)	ID NO:1132)	ID NO:1272)	ID NO:1414)
DTSRHTS (SEQ	DTFRHTS (SEQ	DTYRHTS (SEQ	DTRRHTS (SEQ	DTMRHTS (SEQ	DTKRHTS (SEQ	DTLRHTS (SEQ
ID NO:585)	ID NO:720)	ID NO:861)	ID NO:998)	ID NO:1133)	ID NO:1273)	ID NO:1415)
DTSRHDS (SEQ	DTFRHDS (SEQ	DTYRHDS (SEQ	DTRRHDS (SEQ	DTMRHDS (SEQ	DTKRHDS (SEQ	DTLRHDS (SEQ
ID NO:586)	ID NO:721)	ID NO:862)	ID NO:999)	ID NO:1134)	ID NO:1274)	ID NO:1416)
DTSRQAS (SEQ	DTFRQAS (SEQ	DTYRQAS (SEQ	DTRRQAS (SEQ	DTMRQAS (SEQ	DTKRQAS (SEQ	DTLRQAS (SEQ
ID NO:587)	ID NO:722)	ID NO:863)	ID NO:1000)	ID NO:1135)	ID NO:1275)	ID NO:1417)
DTSRQSS (SEQ	DTFRQSS (SEQ	DTYRQSS (SEQ	DTRRQSS (SEQ	DTMRQSS (SEQ	DTKRQSS (SEQ	DTLRQSS (SEQ
ID NO:588)	ID NO:723)	ID NO:864)	ID NO:1001)	ID NO:1136)	ID NO:1276)	ID NO:1418)
DTSRQKS (SEQ	DTFRQKS (SEQ	DTYRQKS (SEQ	DTRRQKS (SEQ	DTMRQKS (SEQ	DTKRQKS (SEQ	DTLRQKS (SEQ
ID NO:589)	ID NO:724)	ID NO:865)	ID NO:1002)	ID NO:1137)	ID NO:1277)	ID NO:1419)
DTSRQRS (SEQ	DTFRQRS (SEQ	DTYRQRS (SEQ	DTRRQRS (SEQ	DTMRQRS (SEQ	DTKRQRS (SEQ	DTLRQRS (SEQ
ID NO:590)	ID NO:725)	ID NO:866)	ID NO:1003)	ID NO:1138)	ID NO:1278)	ID NO:1420)
DTSRQHS (SEQ	DTFRQHS (SEQ	DTYRQHS (SEQ	DTRRQHS (SEQ	DTMRQHS (SEQ	DTKRQHS (SEQ	DTLRQHS (SEQ
ID NO:591)	ID NO:726)	ID NO:867)	ID NO:1004)	ID NO:1139)	ID NO:1279)	ID NO:1421)
DTSRQPS (SEQ	DTFRQPS (SEQ	DTYRQPS (SEQ	DTRRQPS (SEQ	DTMRQPS (SEQ	DTKRQPS (SEQ	DTLRQPS (SEQ
ID NO:592)	ID NO:727)	ID NO:868)	ID NO:1005)	ID NO:1140)	ID NO:1280)	ID NO:1422)
DTSRQTS (SEQ	DTFRQTS (SEQ	DTYRQTS (SEQ	DTRRQTS (SEQ	DTMRQTS (SEQ	DTKRQTS (SEQ	DTLRQTS (SEQ
ID NO:593)	ID NO:728)	ID NO:869)	ID NO:1006)	ID NO:1141)	ID NO:1281)	ID NQ:1423)
DTSRQDS (SEQ	DTFRQDS (SEQ	DTYRQDS (SEQ	DTRRQDS (SEQ	DTMRQDS (SEQ	DTKRQDS (SEQ	DTLRQDS (SEQ
ID NO:594)	ID NO:729)	ID NO:870)	ID NO:1007)	ID NO:1142)	ID NO:1282)	ID NO:1424)
DTSYLAS (SEQ	DTFYLAS (SEQ	DTYYLAS (SEQ	DTRYLAS (SEQ	DTMYLAS (SEQ	DTKYLAS (SEQ	DTLYLAS (SEQ
ID NOS:	ID NO:99)	ID NO:871)	ID NO:178)	ID NO:158)	ID NO:1283)	ID NO:1425)
81&143)
DTSYLSS (SEQ	DTFYLSS (SEQ	DTYYLSS (SEQ	DTRYLSS (SEQ	DTMYLSS (SEQ	DTKYLSS (SEQ	DTLYLSS (SEQ
ID NOS:	ID NO:90)	ID NO:872)	ID NO:59)	ID NO:160)	ID NO:1284)	ID NO:1426)
85&145)
DTSYLKS (SEQ	DTFYLKS (SEQ	DTYYLKS (SEQ	DTRYLKS (SEQ	DTMYLKS (SEQ	DTKYLKS (SEQ	DTLYLKS (SEQ
ID NO:595)	ID NO:730)	ID NO:873)	ID NO:1008)	ID NO:1143)	ID NO:1285)	ID NO:1427)
DTSYLRS (SEQ	DTFYLRS (SEQ	DTYYLRS (SEQ	DTRYLRS (SEQ	DTMYLRS (SEQ	DTKYLRS (SEQ	DTLYLRS (SEQ
ID NO:596)	ID NO:731)	ID NO:874)	ID NO:1009)	ID NO:1144)	ID NO:1286)	ID NO:1428)
DTSYLHS (SEQ	DTFYLHS (SEQ	DTYYLHS (SEQ	DTRYLHS (SEQ	DTMYLHS (SEQ	DTKYLHS (SEQ	DTLYLHS (SEQ
ID NO:597)	ID NO:732)	ID NO:875)	ID NO:1010)	ID NO:1145)	ID NO:1287)	ID NO:1429)
DTSYLPS (SEQ	DTFYLPS (SEQ	DTYYLPS (SEQ	DTRYLPS (SEQ	DTMYLPS (SEQ	DTKYLPS (SEQ	DTLYLPS (SEQ
ID NO:598)	ID NO:733)	ID NO:876)	ID NO:1011)	ID NO:1146)	ID NO:1288)	ID NO:1430)
DTSYLTS (SEQ	DTFYLTS (SEQ	DTYYLTS (SEQ	DTRYLTS (SEQ	DTMYLTS (SEQ	DTKYLTS (SEQ	DTLYLTS (SEQ
ID NO:599)	ID NO:734)	ID NO:877)	ID NO:1012)	ID NO:1147)	ID NO:1289)	ID NO:1431)
DTSYLDS (SEQ	DTFYLDS (SEQ	DTYYLDS (SEQ	DTRYLDS (SEQ	DTMYLDS (SEQ	DTKYLDS (SEQ	DTLYLDS (SEQ
ID NO:600)	ID NO:735)	ID NO:878)	ID NO:1013)	ID NO:1148)	ID NO:1290)	ID NO:1432)
DTSYHAS (SEQ	DTFYHAS (SEQ	DTYYHAS (SEQ	DTRYHAS (SEQ	DTMYHAS (SEQ	DTKYHAS (SEQ	DTLYHAS (SEQ
ID NO:601)	ID NO:736)	ID NO:879)	ID NO:1014)	ID NO:1149)	ID NO:1291)	ID NO:1433)
DTSYHSS (SEQ	DTFYHSS (SEQ	DTYYHSS (SEQ	DTRYHSS (SEQ	DTMYHSS (SEQ	DTKYHSS (SEQ	DTLYHSS (SEQ
ID NO:602)	ID NO:737)	ID NO:880)	ID NO:1015)	ID NO:1150)	ID NO:1292)	ID NO:1434)
DTSYHKS (SEQ	DTFYHKS (SEQ	DTYYHKS (SEQ	DTRYHKS (SEQ	DTMYHKS (SEQ	DTKYHKS (SEQ	DTLYHKS (SEQ
ID NO:603)	ID NO:738)	ID NO:881)	ID NO:1016)	ID NO:1151)	ID NO:1293)	ID NO:1435)
DTSYHRS (SEQ	DTFYHRS (SEQ	DTYYHRS (SEQ	DTRYHRS (SEQ	DTMYHRS (SEQ	DTKYHRS (SEQ	DTLYHRS (SEQ
ID NO:604)	ID NO:739)	ID NO:882)	ID NO:1017)	ID NO:1152)	ID NO:1294)	ID NO:1436)
DTSYHHS (SEQ	DTFYHHS (SEQ	DTYYHHS (SEQ	DTRYHHS (SEQ	DTMYHHS (SEQ	DTKYHHS (SEQ	DTLYHHS (SEQ
ID NO:605)	ID NO:740)	ID NO:883)	ID NO:1018)	ID NO:1153)	ID NO:1295)	ID NO:1437)
DTSYHPS (SEQ	DTFYHPS (SEQ	DTYYHPS (SEQ	DTRYHPS (SEQ	DTMYHPS (SEQ	DTKYHPS (SEQ	DTLYHPS (SEQ
ID NO:606)	ID NO:741)	ID NO:884)	ID NO:1019)	ID NO:1154)	ID NO:1296)	ID NO:1438)
DTSYHTS (SEQ	DTFYHTS (SEQ	DTYYHTS (SEQ	DTRYHTS (SEQ	DTMYHTS (SEQ	DTKYHTS (SEQ	DTLYHTS (SEQ
ID NO:607)	ID NO:742)	ID NO:885)	ID NO:1020)	ID NO:1155)	ID NO:1297)	ID NO:1439)
DTSYHDS (SEQ	DTFYHDS (SEQ	DTYYHDS (SEQ	DTRYHDS (SEQ	DTMYHDS (SEQ	DTKYHDS (SEQ	DTLYHDS (SEQ
ID NO:608)	ID NO:743)	ID NO:886)	ID NO:1021)	ID NO:1156)	ID NO:1298)	ID NO:1440)
DTSYQAS (SEQ	DTFYQAS (SEQ	DTYYQAS (SEQ	DTRYQAS (SEQ	DTMYQAS (SEQ	DTKYQAS (SEQ	DTLYQAS (SEQ
ID NO:147)	ID NO:744)	ID NO:887)	ID NO:167)	ID NO:151)	ID NO:1299)	ID NO:1441)
DTSYQSS (SEQ	DTFYQSS (SEQ	DTYYQSS (SEQ	DTRYQSS (SEQ	DTMYQSS (SEQ	DTKYQSS (SEQ	DTLYQSS (SEQ
ID NO:149)	ID NO:745)	ID NO:888)	ID NO:53)	ID NO:43)	ID NO:1300)	ID NO:1442)
DTSYQKS (SEQ	DTFYQKS (SEQ	DTYYQKS (SEQ	DTRYQKS (SEQ	DTMYQKS (SEQ	DTKYQKS (SEQ	DTLYQKS (SEQ
ID NO:609)	ID NO:746)	ID NO:889)	ID NO:1022)	ID NO:1157)	ID NO:1301)	ID NO:1443)
DTSYQRS (SEQ	DTFYQRS (SEQ	DTYYQRS (SEQ	DTRYQRS (SEQ	DTMYQRS (SEQ	DTKYQRS (SEQ	DTLYQRS (SEQ
ID NO:610)	ID NO:747)	ID NO:890)	ID NO:1023)	ID NO:1158)	ID NO:1302)	ID NO:1444)
DTSYQHS (SEQ	DTFYQHS (SEQ	DTYYQHS (SEQ	DTRYQHS (SEQ	DTMYQHS (SEQ	DTKYQHS (SEQ	DTLYQHS (SEQ
ID NO:611)	ID NO:748)	ID NO:891)	ID NO:1024)	ID NO:1159)	ID NO:1303)	ID NO:1445)
DTSYQPS (SEQ	DTFYQPS (SEQ	DTYYQPS (SEQ	DTRYQPS (SEQ	DTMYQPS (SEQ	DTKYQPS (SEQ	DTLYQPS (SEQ
ID NO:612)	ID NO:749)	ID NO:892)	ID NO:1025)	ID NO:1160)	ID NO:1304)	ID NO:1446)
DTSYQTS (SEQ	DTFYQTS (SEQ	DTYYQTS (SEQ	DTRYQTS (SEQ	DTMYQTS (SEQ	DTKYQTS (SEQ	DTLYQTS (SEQ
ID NO:613)	ID NO:750)	ID NO:893)	ID NO:1026)	ID NO:1161)	ID NO:1305)	ID NO:1447)
DTSYQDS (SEQ	DTFYQDS (SEQ	DTYYQDS (SEQ	DTRYQDS (SEQ	DTMYQDS (SEQ	DTKYQDS (SEQ	DTLYQDS (SEQ
ID NO:614)	ID NO:751)	ID NO:894)	ID NO:1027)	ID NO:1162)	ID NO:1306)	ID NO:1448)
DTSFLAS (SEQ	DTFFLAS (SEQ	DTYFLAS (SEQ	DTRFLAS (SEQ	DTMFLAS (SEQ	DTKFLAS (SEQ	DTLFLAS (SEQ
ID NO:615)	ID NO:752)	ID NO:895)	ID NO:1028)	ID NO:1163)	ID NO:1307)	ID NO:1449)
DTSFLSS (SEQ	DTFFLSS (SEQ	DTYFLSS (SEQ	DTRFLSS (SEQ	DTMFLSS (SEQ	DTKFLSS (SEQ	DTLFLSS (SEQ
ID NO:616)	ID NO:753)	ID NO:896)	ID NO:1029)	ID NO:1164)	ID NO:1308)	ID NO:1450)
DTSFLKS (SEQ	DTFFLKS (SEQ	DTYFLKS (SEQ	DTRFLKS (SEQ	DTMFLKS (SEQ	DTKFLKS (SEQ	DTLFLKS (SEQ
ID NO:617)	ID NO:754)	ID NO:897)	ID NO:1030)	ID NO:1165)	ID NO:1309)	ID NO:1451)
DTSFLRS (SEQ	DTFFLRS (SEQ	DTYFLRS (SEQ	DTRFLRS (SEQ	DTMFLRS (SEQ	DTKFLRS (SEQ	DTLFLRS (SEQ
ID NO:618)	ID NO:755)	ID NO:898)	ID NO:1031)	ID NO:1166)	ID NO:1310)	ID NO:1452)
DTSFLHS (SEQ	DTFFLHS (SEQ	DTYFLHS (SEQ	DTRFLHS (SEQ	DTMFLHS (SEQ	DTKFLHS (SEQ	DTLFLHS (SEQ
ID NO:619)	ID NO:756)	ID NO:899)	ID NO:1032)	ID NO:1167)	ID NO:1311)	ID NO:1453)
DTSFLPS (SEQ	DTFFLPS (SEQ	DTYFLPS (SEQ	DTRFLPS (SEQ	DTMFLPS (SEQ	DTKFLPS (SEQ	DTLFLPS (SEQ
ID NO:620)	ID NO:757)	ID NO:900)	ID NO:1033)	ID NO:1168)	ID NO:1312)	ID NO:1454)
DTSFLTS (SEQ	DTFFLTS (SEQ	DTYFLTS (SEQ	DTRFLTS (SEQ	DTMFLTS (SEQ	DTKFLTS (SEQ	DTLFLTS (SEQ
ID NO:621)	ID NO:758)	ID NO:901)	ID NO:1034)	ID NO:1169)	ID NO:1313)	ID NO:1455)
DTSFLDS (SEQ	DTFFLDS (SEQ	DTYFLDS (SEQ	DTRFLDS (SEQ	DTMFLDS (SEQ	DTKFLDS (SEQ	DTLFLDS (SEQ
ID NO:77)	ID NO:50)	ID NO:902)	ID NO:1035)	ID NO:1170)	ID NO:1314)	ID NO:1456)
DTSFHAS (SEQ	DTFFHAS (SEQ	DTYFHAS (SEQ	DTRFHAS (SEQ	DTMFHAS (SEQ	DTKFHAS (SEQ	DTLFHAS (SEQ
ID NO:622)	ID NO:759)	ID NO:903)	ID NO:1036)	ID NO:1171)	ID NO:1315)	ID NO:1457)
DTSFHSS (SEQ	DTFFHSS (SEQ	DTYFHSS (SEQ	DTRFHSS (SEQ	DTMFHSS (SEQ	DTKFHSS (SEQ	DTLFHSS (SEQ
ID NO:623)	ID NO:760)	ID NO:904)	ID NO:1037)	ID NO:1172)	ID NO:1316)	ID NO:1458)
DTSFHKS (SEQ	DTFFHKS (SEQ	DTYFHKS (SEQ	DTRFHKS (SEQ	DTMFHKS (SEQ	DTKFHKS (SEQ	DTLFHKS (SEQ
ID NO:624)	ID NO:761)	ID NO:905)	ID NO:1038)	ID NO:1173)	ID NO:1317)	ID NO:1459)
DTSFHRS (SEQ	DTFFHRS (SEQ	DTYFHRS (SEQ	DTRFHRS (SEQ	DTMFHRS (SEQ	DTKFHRS (SEQ	DTLFHRS (SEQ
ID NO:625)	ID NO:762)	ID NO:906)	ID NO:1039)	ID NO:1174)	ID NO:1318)	ID NO:1460)
DTSFHHS (SEQ	DTFFHHS (SEQ	DTYFHHS (SEQ	DTRFHHS (SEQ	DTMFHHS (SEQ	DTKFHHS (SEQ	DTLFHHS (SEQ
ID NO:626)	ID NO:763)	ID NO:907)	ID NO:1040)	ID NO:1175)	ID NO:1319)	ID NO:1461)
DTSFHPS (SEQ	DTFFHPS (SEQ	DTYFHPS (SEQ	DTRFHPS (SEQ	DTMFHPS (SEQ	DTKFHPS (SEQ	DTLFHPS (SEQ
ID NO:627)	ID NO:764)	ID NO:908)	ID NO:1041)	ID NO:1176)	ID NO:1320)	ID NO:1462)
DTSFHTS (SEQ	DTFFHTS (SEQ	DTYFHTS (SEQ	DTRFHTS (SEQ	DTMFHTS (SEQ	DTKFHTS (SEQ	DTLFHTS (SEQ
ID NO:628)	ID NO:765)	ID NO:909)	ID NO:1042)	ID NO:1177)	ID NO:1321)	ID NO:1463)
DTSFHDS (SEQ	DTFFHDS (SEQ	DTYFHDS (SEQ	DTRFHDS (SEQ	DTMFHDS (SEQ	DTKFHDS (SEQ	DTLFHDS (SEQ
ID NO:629)	ID NO:766)	ID NO:910)	ID NO:1043)	ID NO:1178)	ID NO:1322)	ID NO:1464)
DTSFQAS (SEQ	DTFFQAS (SEQ	DTYFQAS (SEQ	DTRFQAS (SEQ	DTMFQAS (SEQ	DTKFQAS (SEQ	DTLFQAS (SEQ
ID NO:630)	ID NO:767)	ID NO:911)	ID NO:1044)	ID NO:1179)	ID NO:1323)	ID NO:1465)
DTSFQSS (SEQ	DTFFQSS (SEQ	DTYFQSS (SEQ	DTRFQSS (SEQ	DTMFQSS (SEQ	DTKFQSS (SEQ	DTLFQSS (SEQ
ID NO:631)	ID NO:768)	ID NO:912)	ID NO:1045)	ID NO:1180)	ID NO:1324)	ID NO:1466)
DTSFQKS (SEQ	DTFFQKS (SEQ	DTYFQKS (SEQ	DTRFQKS (SEQ	DTMFQKS (SEQ	DTKFQKS (SEQ	DTLFQKS (SEQ
ID NO:632)	ID NO:769)	ID NO:913)	ID NO:1046)	ID NO:1181)	ID NO:1325)	ID NO:1467)
DTSFQRS (SEQ	DTFFQRS (SEQ	DTYFQRS (SEQ	DTRFQRS (SEQ	DTMFQRS (SEQ	DTKFQRS (SEQ	DTLFQRS (SEQ
ID NO:633)	ID NO:770)	ID NO:914)	ID NO:1047)	ID NO:1182)	ID NO:1326)	ID NO:1468)
DTSFQHS (SEQ	DTFFQHS (SEQ	DTYFQHS (SEQ	DTRFQHS (SEQ	DTMFQHS (SEQ	DTKFQHS (SEQ	DTLFQHS (SEQ
ID NO:634)	ID NO:771)	ID NO:915)	ID NO:1048)	ID NO:1183)	ID NO:1327)	ID NO:1469)
DTSFQPS (SEQ	DTFFQPS (SEQ	DTYFQPS (SEQ	DTRFQPS (SEQ	DTMFQPS (SEQ	DTKFQPS (SEQ	DTLFQPS (SEQ
ID NO:635)	ID NO:772)	ID NO:916)	ID NO:1049)	ID NO:1184)	ID NO:1328)	ID NO:1470)
DTSFQTS (SEQ	DTFFQTS (SEQ	DTYFQTS (SEQ	DTRFQTS (SEQ	DTMFQTS (SEQ	DTKFQTS (SEQ	DTLFQTS (SEQ
ID NO:636)	ID NO:773)	ID NO:917)	ID NO:1050)	ID NO:1185)	ID NO:1329)	ID NO:1471)
DTSFQDS (SEQ	DTFFQDS (SEQ	DTYFQDS (SEQ	DTRFQDS (SEQ	DTMFQDS (SEQ	DTKFQDS (SEQ	DTLFQDS (SEQ
ID NO:637)	ID NO:774)	ID NO:918)	ID NO:1051)	ID NO:1186)	ID NO:1330)	ID NO:1472)
DTSLLAS (SEQ	DTFLLAS (SEQ	DTYLLAS (SEQ	DTRLLAS (SEQ	DTMLLAS (SEQ	DTKLLAS (SEQ	DTLLLAS (SEQ
ID NO:124)	ID NO:775)	ID NO:919)	ID NO:1052)	ID NO:1187)	ID NO:1331)	ID NO:133)
DTSLLSS (SEQ	DTFLLSS (SEQ	DTYLLSS (SEQ	DTRLLSS (SEQ	DTMLLSS (SEQ	DTKLLSS (SEQ	DTLLLSS (SEQ
ID NO:638)	ID NO:776)	ID NO:920)	ID NO:1053)	ID NO:1188)	ID NO:1332)	ID NO:1473)
DTSLLKS (SEQ	DTFLLKS (SEQ	DTYLLKS (SEQ	DTRLLKS (SEQ	DTMLLKS (SEQ	DTKLLKS (SEQ	DTLLLKS (SEQ
ID NO:639)	ID NO:777)	ID NO:921)	ID NO:1054)	ID NO:1189)	ID NO:1333)	ID NO:1474)
DTSLLRS (SEQ	DTFLLRS (SEQ	DTYLLRS (SEQ	DTRLLRS (SEQ	DTMLLRS (SEQ	DTKLLRS (SEQ	DTLLLRS (SEQ
ID NO:640)	ID NO:778)	ID NO:922)	ID NO:1055)	ID NO:1190)	ID NO:1334)	ID NO:1475)
DTSLLHS (SEQ	DTFLLHS (SEQ	DTYLLHS (SEQ	DTRLLHS (SEQ	DTMLLHS (SEQ	DTKLLHS (SEQ	DTLLLHS (SEQ
ID NO:641)	ID NO:779)	ID NO:923)	ID NO:1056)	ID NO:1191)	ID NO:1335)	ID NO:1476)
DTSLLPS (SEQ	DTFLLPS (SEQ	DTYLLPS (SEQ	DTRLLPS (SEQ	DTMLLPS (SEQ	DTKLLPS (SEQ	DTLLLPS (SEQ
ID NO:642)	ID NO:780)	ID NO:924)	ID NO:1057)	ID NO:1192)	ID NO:1336)	ID NO:1477)
DTSLLTS (SEQ	DTFLLTS (SEQ	DTYLLTS (SEQ	DTRLLTS (SEQ	DTMLLTS (SEQ	DTKLLTS (SEQ	DTLLLTS (SEQ
ID NO:643)	ID NO:781)	ID NO:925)	ID NO:1058)	ID NO:1193)	ID NO:1337)	ID NO:1478)
DTSLLDS (SEQ	DTFLLDS (SEQ	DTYLLDS (SEQ	DTRLLDS (SEQ	DTMLLDS (SEQ	DTKLLDS (SEQ	DTLLLDS (SEQ
ID NO:126)	ID NO:782)	ID NO:926)	ID NO:1059)	ID NO:1194)	ID NO:1338)	ID NO:75)
DTSLHAS (SEQ	DTFLHAS (SEQ	DTYLHAS (SEQ	DTRLHAS (SEQ	DTMLHAS (SEQ	DTKLHAS (SEQ	DTLLHAS (SEQ
ID NO:644)	ID NO:783)	ID NO:927)	ID NO:1060)	ID NO:1195)	ID NO:1339)	ID NO:1479)
DTSLHSS (SEQ	DTFLHSS (SEQ	DTYLHSS (SEQ	DTRLHSS (SEQ	DTMLHSS (SEQ	DTKLHSS (SEQ	DTLLHSS (SEQ
ID NO:645)	ID NO:784)	ID NO:928)	ID NO:1061)	ID NO:1196)	ID NO:1340)	ID NO:1480)
DTSLHKS (SEQ	DTFLHKS (SEQ	DTYLHKS (SEQ	DTRLHKS (SEQ	DTMLHKS (SEQ	DTKLHKS (SEQ	DTLLHKS (SEQ
ID NO:646)	ID NO:785)	ID NO:929)	ID NO:1062)	ID NO:1197)	ID NO:1341)	ID NO:1481)
DTSLHRS (SEQ	DTFLHRS (SEQ	DTYLHRS (SEQ	DTRLHRS (SEQ	DTMLHRS (SEQ	DTKLHRS (SEQ	DTLLHRS (SEQ
ID NO:647)	ID NO:786)	ID NO:930)	ID NO:1063)	ID NO:1198)	ID NO:1342)	ID NO:1482)
DTSLHHS (SEQ	DTFLHHS (SEQ	DTYLHHS (SEQ	DTRLHHS (SEQ	DTMLHHS (SEQ	DTKLHHS (SEQ	DTLLHHS (SEQ
ID NO:648)	ID NO:787)	ID NO:931)	ID NO:1064)	ID NO:1199)	ID NO:1343)	ID NO:1483)
DTSLHPS (SEQ	DTFLHPS (SEQ	DTYLHPS (SEQ	DTRLHPS (SEQ	DTMLHPS (SEQ	DTKLHPS (SEQ	DTLLHPS (SEQ
ID NO:649)	ID NO:788)	ID NO:932)	ID NO:1065)	ID NO:1200)	ID NO:1344)	ID NO:1484)
DTSLHTS (SEQ	DTFLHTS (SEQ	DTYLHTS (SEQ	DTRLHTS (SEQ	DTMLHTS (SEQ	DTKLHTS (SEQ	DTLLHTS (SEQ
ID NO:650)	ID NO:789)	ID NO:933)	ID NO:1066)	ID NO:1201)	ID NO:1345)	ID NO:1485)
DTSLHDS (SEQ	DTFLHDS (SEQ	DTYLHDS (SEQ	DTRLHDS (SEQ	DTMLHDS (SEQ	DTKLHDS (SEQ	DTLLHDS (SEQ
ID NO:651)	ID NO:790)	ID NO:934)	ID NO:1067)	ID NO:1202)	ID NO:1346)	ID NO:1486)
DTSLQAS (SEQ	DTFLQAS (SEQ	DTYLQAS (SEQ	DTRLQAS (SEQ	DTMLQAS (SEQ	DTKLQAS (SEQ	DTLLQAS (SEQ
ID NO:652)	ID NO:791)	ID NO:935)	ID NO:1068)	ID NO:1203)	ID NO:1347)	ID NO:1487)
DTSLQSS (SEQ	DTFLQSS (SEQ	DTYLQSS (SEQ	DTRLQSS (SEQ	DTMLQSS (SEQ	DTKLQSS (SEQ	DTLLQSS (SEQ
ID NO:653)	ID NO:792)	ID NO:936)	ID NO:1069)	ID NO:1204)	ID NO:1348)	ID NO:1488)
DTSLQKS (SEQ	DTFLQKS (SEQ	DTYLQKS (SEQ	DTRLQKS (SEQ	DTMLQKS (SEQ	DTKLQKS (SEQ	DTLLQKS (SEQ
ID NO:654)	ID NO:793)	ID NO:937)	ID NO:1070)	ID NO:1205)	ID NO:1349)	ID NO:1489)
DTSLQRS (SEQ	DTFLQRS (SEQ	DTYLQRS (SEQ	DTRLQRS (SEQ	DTMLQRS (SEQ	DTKLQRS (SEQ	DTLLQRS (SEQ
ID NO:655)	ID NO:794)	ID NO:938)	ID NO:1071)	ID NO:1206)	ID NO:1350)	ID NO:1490)
DTSLQHS (SEQ	DTFLQHS (SEQ	DTYLQHS (SEQ	DTRLQHS (SEQ	DTMLQHS (SEQ	DTKLQHS (SEQ	DTLLQHS (SEQ
ID NO:656)	ID NO:795)	ID NO:939)	ID NO:1072)	ID NO:1207)	ID NO:1351)	ID NO:1491)
DTSLQPS (SEQ	DTFLQPS (SEQ	DTYLQPS (SEQ	DTRLQPS (SEQ	DTMLQPS (SEQ	DTKLQPS (SEQ	DTLLQPS (SEQ
ID NO:657)	ID NO:796)	ID NO:940)	ID NO:1073)	ID NO:1208)	ID NO:1352)	ID NO:1492)
DTSLQTS (SEQ	DTFLQTS (SEQ	DTYLQTS (SEQ	DTRLQTS (SEQ	DTMLQTS (SEQ	DTKLQTS (SEQ	DTLLQTS (SEQ
ID NO:658)	ID NO:797)	ID NO:941)	ID NO:1074)	ID NO:1209)	ID NO:1353)	ID NO:1493)
DTSLQDS (SEQ	DTFLQDS (SEQ	DTYLQDS (SEQ	DTRLQDS (SEQ	DTMLQDS (SEQ	DTKLQDS (SEQ	DTLLQDS (SEQ
ID NO:659)	ID NO:798)	ID NO:942)	ID NO:1075)	ID NO:1210)	ID NO:1354)	ID NO:1494)
Bold faced & underlined amino acid residues are the residues which differ from the amino acid sequence in palivizumab