RU2847808C9 - Neutralising respiratory syncytial virus antibody and its application - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: group of inventions relates to biotechnology and antibody-based medicines. An antibody specifically binding to respiratory syncytial virus (RSV), a nucleic acid molecule encoding the antibody, the use of the antibody in the manufacture of a medicinal product for the prevention or treatment of RSV-induced disease, and a method for the prevention or treatment of RSV-induced disease are proposed.

EFFECT: antibody that binds to the F protein of human RSV, has high activity, and is capable of successfully neutralising clinical strains of RSV.

16 cl, 7 dwg, 4 tbl, 4 ex

Description

REFERENCE TO A RELATED APPLICATION

This application claims the benefit of priority from Chinese Patent Application No. 202210871810.9, filed on July 22, 2022, and incorporated herein by reference in its entirety.

AREA OF TECHNOLOGY

This application broadly relates to genetic engineering and antibody-based medicinal products. Specifically, this application relates to an antibody to the respiratory syncytial virus (RSV) and the use of this antibody in the prevention or treatment of RSV-related disease.

LEVEL OF TECHNOLOGY

Large, high-quality antibody libraries are an important source of molecules for screening high-affinity antibodies to any antigen and possess high commercial and practical value. Currently, the majority of high-quality antibody libraries reported by domestic and international sources are generated using electrotransformation technology. However, the size of antibody libraries generated by electrotransformation is limited due to insufficient transformation efficiency. The process of generating antibody libraries is periodic and involves multiple batches, making it time-consuming and requiring significant human and material resources.

Respiratory syncytial virus (RSV) is the most dangerous viral pathogen causing acute lower respiratory tract infections (ALRI) in children under 5 years of age worldwide ¹ . RSV infection is the leading cause of hospitalization of infants and young children with respiratory viral infections and poses a serious health risk to children, especially those born prematurely, with congenital heart defects, or with primary immunodeficiencies. The top five countries with the highest incidence of RSV infection are Pakistan, India, Nigeria, China, and Indonesia, which account for almost half of the cases of RSV-associated ALRI worldwide ² .

Respiratory syncytial virus (RSV) belongs to the Pneumoviridae family, Orthopneumovirus genus, and is a virus with a nonsegmented genome containing single-stranded RNA of negative polarity ³ . It can be classified into two subtypes, A and B, depending on differences in surface antigens ⁴ . The RSV genome contains 10 genes encoding 11 proteins. The adhesive G protein and the fusogenic F protein are the main protective antigens on the virus surface, which can induce the production of neutralizing antibodies by the body ⁵ . The G protein is characterized by a high degree of subtype differentiation, therefore, most antibodies to the G protein are subtype-specific antibodies ⁶ . The F protein is characterized by a high degree of subtype conservation, therefore, antibodies induced by the F protein are capable of simultaneously inhibiting RSV types A and B.

The F protein is a type I transmembrane protein, with its inactive precursor (F0) consisting of 574 amino acids ⁷ . Three F0s form a trimer. Host furin cleaves F0 between amino acids at positions 109 and 110 and between amino acids at positions 136 and 137 in F0 during its passage through the Golgi. After cleavage, a short peptide of 27 amino acids in the middle (P27) is released, while the remaining two segments F2 and F1 form the active F protein through disulfide bonds (Cys69–Cys212 and Cys37–Cys439) ⁸ . A highly hydrophobic fusion peptide (FP) is presented at the N-terminus of the F1 protein and is located in the hydrophobic cavity of the protein, thereby being protected from the effects of the external hydrophilic environment. When the F protein is presented on the virion or cell surface, it is not in a structurally stable form, but in a highly metastable pre-fusion glyprotein (Pre-F) form. ⁹ Subsequently, a series of significant structural changes occur in the N-terminus of the F protein, inducing membrane fusion and, consequently, viral infection of the cell. This process also leads to the conversion of the F protein from the highly metastable Pre-F structure to the stable post-fusion glyprotein (Post-F) form.

Currently, there is no vaccine to prevent RSV infection. Palivizumab (brand name: Synagis) is approved by the U.S. Food and Drug Administration for the prevention of RSV infection in high-risk infants and young children. Typically, up to five injections are required per RSV season, which is costly and makes it impossible to provide widespread coverage to infants and young children.

Given clinical needs, the development of antibodies to respiratory syncytial virus is important from a medical point of view for the prevention of diseases caused by RSV infection.

ESSENCE OF THE INVENTION

According to a first aspect, the present application provides an antibody to respiratory syncytial virus (RSV), comprising a heavy chain variable region comprising heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2 and HCDR3, and a light chain variable region comprising light chain complementarity determining regions (LCDRs) LCDR1, LCDR2 and LCDR3, wherein

the amino acid sequence of HCDR1 is presented in SEQ ID NO:32, the amino acid sequence of HCDR2 is presented in SEQ ID NO:33, the amino acid sequence of HCDR3 is presented in SEQ ID NO:34, the amino acid sequence of LCDR1 is presented in SEQ ID NO:35, the amino acid sequence of LCDR2 is presented in SEQ ID NO:36, and the amino acid sequence of LCDR3 is presented in SEQ ID NO:37; or

the amino acid sequence of HCDR1 is presented in SEQ ID NO:38, the amino acid sequence of HCDR2 is presented in SEQ ID NO:39, the amino acid sequence of HCDR3 is presented in SEQ ID NO:40, the amino acid sequence of LCDR1 is presented in SEQ ID NO:35, the amino acid sequence of LCDR2 is presented in SEQ ID NO:36, and the amino acid sequence of LCDR3 is presented in SEQ ID NO:43;

wherein the amino acid sequences of the complementarity-determining regions of the heavy chain and the complementarity-determining regions of the light chain are determined according to the Cabot nomenclature.

In some embodiments of the first aspect, the amino acid sequence of the heavy chain variable region of the antibody is shown in SEQ ID NO: 28 or 30.

In some embodiments of the first aspect, the amino acid sequence of the light chain variable region of the antibody is shown in SEQ ID NO: 29 or 31.

In some embodiments of the first aspect, the amino acid sequence of the heavy chain variable region of the antibody is shown in SEQ ID NO: 28, wherein the amino acid sequence of the light chain variable region of the antibody is shown in SEQ ID NO: 29; or

the amino acid sequence of the variable region of the heavy chain of the antibody is presented in SEQ ID NO: 30, while the amino acid sequence of the variable region of the light chain of the antibody is presented in SEQ ID NO: 31.

According to a second aspect, the present application provides an antibody to respiratory syncytial virus (RSV), wherein the amino acid sequence of the heavy chain variable region of the antibody is at least 90% identical to the amino acid sequence defined in SEQ ID NO: 28 or 30, while the amino acid sequence of the light chain variable region of the antibody is at least 90% identical to the amino acid sequence defined in SEQ ID NO: 29 or 31.

In some embodiments of the first or second aspects, the antibody is a neutralizing antibody.

In some embodiments of the first or second aspects, the antibody may be capable of binding to the F protein of human respiratory syncytial virus (RSV).

In some embodiments of the first or second aspects, the antibody is a Fab fragment, a whole antibody, an F(ab') ₂ fragment, or a single-chain Fv fragment (scFv fragment).

In some embodiments of the first or second aspects, the antibody is a monoclonal antibody.

In some embodiments of the first or second aspects, the antibody comprises a heavy chain constant region of an IgG1 subtype, an IgG2 subtype, or an IgG4 subtype.

In some embodiments of the first or second aspects, the heavy chain constant region comprises an Fc region of an IgG1 subtype heavy chain constant region, wherein the amino acids at positions 252, 254, 256 of the Fc region sequence are Y, T and E, respectively, wherein the amino acid positions of the antibody constant region are determined according to EU numbering.

In some embodiments of the first or second aspects, the antibody comprises a light chain constant region of the kappa subtype or the lambda subtype.

According to a third aspect, the present application provides a nucleic acid molecule encoding an antibody according to the first or second aspect.

According to a fourth aspect, the present application provides a pharmaceutical composition comprising an antibody according to the first or second aspect and a pharmaceutically acceptable excipient, diluent or carrier.

According to a fifth aspect, the present application provides the use of an antibody according to the first or second aspect, a nucleic acid molecule according to the third aspect, or a pharmaceutical composition according to the fourth aspect in the manufacture of a medicament for the prevention or treatment of a disease caused by RSV.

According to a sixth aspect, the present application provides a method for preventing or treating a disease caused by RSV, comprising the step of administering to a subject in need thereof an antibody according to the first or second aspect or a pharmaceutical composition according to the fourth aspect.

According to a seventh aspect, the present application provides a combination of plasmids for expressing a library of Fab fragments of antibodies, comprising a first plasmid and a second plasmid, wherein

the first plasmid contains the VH-CH1 expression element of the antibody heavy chain and the first recombination site;

the second plasmid contains the VH-CH1 expression element of the antibody light chain and the second recombination site; wherein

the first recombination site is different from the second recombination site.

In some embodiments of the seventh aspect, the combination of plasmids comprises a plurality of first plasmids, wherein the plurality of first plasmids carry different VH-CH1 antibody heavy chain expression elements and have an identical first recombination site.

In some embodiments of the seventh aspect, the plasmid combination comprises a plurality of second plasmids, wherein the plurality of second plasmids carry different VH-CH1 antibody light chain expression elements and have an identical second recombination site.

In some embodiments of the seventh aspect, the first plasmid and/or the second plasmid is a phagemid.

In some embodiments of the seventh aspect, the first plasmid and the second plasmid have different origins of replication.

In some embodiments of the seventh aspect, the origin of replication is selected from the group consisting of one or more of the pBR plasmid origin of replication (pBR origin of replication), the CDF origin of replication, the M13 filamentous phage origin of replication (f1 origin of replication), and the p15A origin of replication.

In some embodiments of the seventh aspect, the first recombination site and the second recombination site are phage integration sites in the genome of the phage and the host bacterium, respectively.

In some embodiments of the seventh aspect, the first recombination site is an attP site, and the second recombination site is an attB site.

In some embodiments of the seventh aspect, the first recombination site and the second recombination site are located at polyclonal enzyme cleavage sites of the first plasmid and the second plasmid, respectively.

In some embodiments of the seventh aspect, the light chain constant region (CL region) of the antibody is a human kappa light chain constant region or a human lambda light chain constant region.

In some embodiments of the seventh aspect, the CH1 domain of the heavy chain constant region of the antibody is of the IgG1, IgG2, IgG3 or IgG4 subtype.

In some embodiments of the seventh aspect, the first plasmid and/or the second plasmid comprises a resistance gene encoding region.

In some embodiments of the seventh aspect, a nucleic acid molecule comprising a nucleic acid molecule encoding the CH1 domain of the constant region of an antibody heavy chain is fused at its 3' end to the 5' end of a nucleic acid molecule encoding the gIII protein of the filamentous phage M13.

In some embodiments of the seventh aspect, the resistance gene is an antibiotic resistance gene.

In some embodiments of the seventh aspect, the resistance gene is selected from the group consisting of: a chloramphenicol resistance gene (CmR gene), an ampicillin resistance gene (Ampr gene), a kanamycin resistance gene (KaR gene), and a tetracycline resistance gene (TetR gene).

According to an eighth aspect, the present application provides a recombinant system for expressing a library of Fab fragments of antibodies, comprising a first plasmid, a second plasmid and a phage integrase-expressing cell; wherein

the first plasmid contains the VH-CH1 expression element of the antibody heavy chain and the first recombination site;

the second plasmid contains the VH-CH1 expression element of the antibody light chain and the second recombination site; wherein

the first recombination site is different from the second recombination site.

In some embodiments of the eighth aspect, the recombinant system comprises a plurality of first plasmids, wherein the plurality of first plasmids carry different VH-CH1 antibody heavy chain expression elements and have an identical first recombination site.

In some embodiments of the eighth aspect, the recombinant system comprises a plurality of second plasmids, wherein the plurality of second plasmids carry different VH-CH1 light chain antibody expression elements and have an identical second recombination site.

In some embodiments of the eighth aspect, the first plasmid and/or the second plasmid is a phagemid.

In some embodiments of the eighth aspect, the first plasmid and the second plasmid have different origins of replication.

In some embodiments of the eighth aspect, the origin of replication is selected from the group consisting of one or more of the pBR origin of replication, the CDF origin of replication, the M13 filamentous phage origin of replication (f1 origin of replication), and the p15A origin of replication.

In some embodiments of the eighth aspect, the first recombination site and the second recombination site are phage integration sites in the genome of the phage and the host bacterium, respectively.

In some embodiments of the eighth aspect, the first recombination site is an attP site, and the second recombination site is an attB site.

In some embodiments of the eighth aspect, the first recombination site and the second recombination site are located at polyclonal enzyme cleavage sites of the first plasmid and the second plasmid, respectively.

In some embodiments of the eighth aspect, the light chain constant region (CL region) of the antibody is a human kappa light chain constant region or a human lambda light chain constant region.

In some embodiments of the eighth aspect, the CH1 domain of the heavy chain constant region of the antibody is of the IgG1, IgG2, IgG3 or IgG4 subtype.

In some embodiments of the eighth aspect, the first plasmid and/or the second plasmid comprises a resistance gene encoding region.

In some embodiments of the eighth aspect, a nucleic acid molecule comprising a nucleic acid molecule encoding the CH1 domain of a heavy chain constant region of an antibody is fused at its 3' end to the 5' end of a nucleic acid molecule encoding the gIII protein of the filamentous phage M13.

In some embodiments of the eighth aspect, the resistance gene is an antibiotic resistance gene.

In some embodiments of the eighth aspect, the resistance gene is selected from the group consisting of: a chloramphenicol resistance gene (CmR gene), an ampicillin resistance gene (Ampr gene), a kanamycin resistance gene (KaR gene), and a tetracycline resistance gene (TetR gene).

In some embodiments of the eighth aspect, the phage integrase is tyrosine integrase.

In some embodiments of the eighth aspect, the phage integrase is phage lambda integrase.

In some embodiments of the eighth aspect, the phage integrase is an inducibly expressed or constitutively expressed integrase.

In some embodiments of the eighth aspect, the phage integrase is an inducibly expressed integrase.

In some embodiments of the eighth aspect, the phage integrase-expressing cell is a prokaryotic cell.

In some embodiments of the eighth aspect, the phage integrase-expressing cell is E. coli .

In some embodiments of the eighth aspect, the phage integrase-expressing cell is a genetically engineered bacterium.

In some embodiments of the eighth aspect, the first plasmid, the second plasmid and the phage integrase-expressing cell exist independently of one another, or at least one of the first plasmid and the second plasmid has been introduced into the phage integrase-expressing cell.

According to a ninth aspect, the present application provides a recombinant cell for expressing a library of Fab fragments of antibodies, comprising a first plasmid, a second plasmid and expressing a phage integrase; wherein

the first plasmid contains the VH-CH1 expression element of the antibody heavy chain and the first recombination site;

the second plasmid contains the VH-CH1 expression element of the antibody light chain and the second recombination site; wherein

the first recombination site is different from the second recombination site.

In some embodiments of the ninth aspect, the recombinant cell comprises a plurality of first plasmids, wherein the plurality of first plasmids carry different VH-CH1 antibody heavy chain expression elements and have an identical first recombination site.

In some embodiments of the ninth aspect, the recombinant cell comprises a plurality of second plasmids, wherein the plurality of second plasmids carry different VH-CH1 light chain antibody expression elements and have an identical second recombination site.

In some embodiments of the ninth aspect, the first plasmid and/or the second plasmid is a phagemid.

In some embodiments of the ninth aspect, the first plasmid and the second plasmid have different origins of replication.

In some embodiments of the ninth aspect, the origin of replication is selected from the group consisting of one or more of the pBR origin of replication, the CDF origin of replication, the filamentous phage M13 origin of replication (f1 origin of replication), and the p15A origin of replication.

In some embodiments of the ninth aspect, the first recombination site and the second recombination site are phage integration sites in the genome of the phage and the host bacterium, respectively.

In some embodiments of the ninth aspect, the first recombination site is an attP site, and the second recombination site is an attB site.

In some embodiments of the ninth aspect, the first recombination site and the second recombination site are located at polyclonal enzyme cleavage sites of the first plasmid and the second plasmid, respectively.

In some embodiments of the ninth aspect, the light chain constant region (CL region) of the antibody is a human kappa light chain constant region or a human lambda light chain constant region.

In some embodiments of the ninth aspect, the CH1 domain of the heavy chain constant region of the antibody is of the IgG1, IgG2, IgG3 or IgG4 subtype.

In some embodiments of the ninth aspect, the first plasmid and/or the second plasmid comprises a resistance gene encoding region.

In some embodiments of the ninth aspect, a nucleic acid molecule comprising a nucleic acid molecule encoding the CH1 domain of a heavy chain constant region of an antibody is fused at its 3' end to the 5' end of a nucleic acid molecule encoding the gIII protein of the filamentous phage M13.

In some embodiments of the ninth aspect, the resistance gene is an antibiotic resistance gene.

In some embodiments of the ninth aspect, the resistance gene is selected from the group consisting of: a chloramphenicol resistance gene (CmR gene), an ampicillin resistance gene (Ampr gene), a kanamycin resistance gene (KaR gene), and a tetracycline resistance gene (TetR gene).

In some embodiments of the ninth aspect, the phage integrase is tyrosine integrase.

In some embodiments of the ninth aspect, the phage integrase is phage lambda integrase.

In some embodiments of the ninth aspect, the phage integrase is an inducibly expressed or constitutively expressed integrase.

In some embodiments of the ninth aspect, the phage integrase is an inducibly expressed integrase.

In some embodiments of the ninth aspect, the recombinant cell is a prokaryotic cell.

In some embodiments of the ninth aspect, the recombinant cell is E. coli .

In some embodiments of the ninth aspect, the recombinant cell is a genetically engineered bacterium.

According to a tenth aspect, the present application provides a method for producing a library of Fab fragments of antibodies, which comprises transferring a first plasmid and a second plasmid into a cell expressing phage integrase; wherein

the first plasmid contains the VH-CH1 expression element of the antibody heavy chain and the first recombination site;

the second plasmid contains the VH-CH1 expression element of the antibody light chain and the second recombination site; wherein

the first recombination site is different from the second recombination site.

In some embodiments of the tenth aspect, a plurality of first plasmids are transferred into a phage integrase-expressing cell, wherein the plurality of first plasmids carry different VH-CH1 antibody heavy chain expression elements and have an identical first recombination site.

In some embodiments of the tenth aspect, a plurality of second plasmids are transferred into a phage integrase-expressing cell, wherein the plurality of second plasmids carry different VH-CH1 antibody light chain expression elements and have an identical second recombination site.

In some embodiments of the tenth aspect, transferring the first plasmid and/or the second plasmid into the phage integrase-expressing cell comprises the steps of:

(1) transfer the first plasmid and/or the second plasmid into a competent cell;

(2) infecting the competent cell obtained in step (1) with a helper phage to construct a phage library; and

(3) The phage library obtained in step (2) is transferred into a cell expressing phage integrase.

In some embodiments of the tenth aspect, the competent cell is a prokaryotic cell.

In some embodiments of the tenth aspect, the competent cell does not express phage integrase.

In some embodiments of the tenth aspect, the helper phage is an M13 helper phage.

In some embodiments of the tenth aspect, the first plasmid and/or the second plasmid is a phagemid.

In some embodiments of the tenth aspect, the first plasmid and the second plasmid have different origins of replication.

In some embodiments of the tenth aspect, the origin of replication is selected from the group consisting of one or more of the pBR origin of replication, the CDF origin of replication, the filamentous phage M13 origin of replication (f1 origin of replication), and the p15A origin of replication.

In some embodiments of the tenth aspect, the first recombination site and the second recombination site are phage integration sites in the genome of the phage and the host bacterium, respectively.

In some embodiments of the tenth aspect, the first recombination site is an attP site, and the second recombination site is an attB site.

In some embodiments of the tenth aspect, the first recombination site and the second recombination site are located at polyclonal enzyme cleavage sites of the first plasmid and the second plasmid, respectively.

In some embodiments of the tenth aspect, the light chain constant region (CL region) of the antibody is a human kappa light chain constant region or a human lambda light chain constant region.

In some embodiments of the tenth aspect, the CH1 domain of the heavy chain constant region of the antibody is of the IgG1, IgG2, IgG3 or IgG4 subtype.

In some embodiments of the tenth aspect, the first plasmid and/or the second plasmid comprises a resistance gene encoding region.

In some embodiments of the tenth aspect, the resistance gene is selected from the group consisting of: a chloramphenicol resistance gene (CmR gene), an ampicillin resistance gene (Ampr gene), a kanamycin resistance gene (KaR gene), and a tetracycline resistance gene (TetR gene).

In some embodiments of the tenth aspect, the phage integrase is tyrosine integrase.

In some embodiments of the tenth aspect, the phage integrase is phage lambda integrase.

In some embodiments of the tenth aspect, the phage integrase is an inducibly expressed or constitutively expressed integrase.

In some embodiments of the tenth aspect, the phage integrase is an inducibly expressed integrase.

In some embodiments of the tenth aspect, the phage integrase-expressing cell is a prokaryotic cell.

In some embodiments of the tenth aspect, the phage integrase-expressing cell is E. coli .

In some embodiments of the tenth aspect, the phage integrase-expressing cell is a genetically engineered bacterium.

According to an eleventh aspect, the present application provides the use of a combination of plasmids according to the seventh aspect, a recombinant system according to the eighth aspect, or a recombinant cell according to the ninth aspect in the production of a library of Fab fragments of antibodies.

DESCRIPTION OF DRAWINGS

Fig. 1 schematically depicts the structure of the phagemid vector pHGDisn-attP-site-new.

Fig. 2 schematically depicts the structure of the prokaryotic expression vector pHKb-attB-site-new.

Fig. 3 shows the results of the ELISA (enzyme-linked immunosorbent assay) assessment of the binding activity of recombinant monoclonal antibodies to the RSV F protein (R3B1h1, R22B1 and positive control antibodies) with RSV-DS-Cav1-A (Insert A) and RSV-DS-Cav1-B (Insert B).

Fig. 4 shows the results of the ELISA assessment of the ability of monoclonal antibodies to the RSV F protein (R3B1h1, R22B1, and clesrovimab) to block the binding of phages to the RSV F protein with RSV-DS-Cav1-B; where Inserts A-C show the results of the assessment of the ability of R3B1h1, R22B1, and clesrovimab to block the binding of purified phage to the RSV F protein with RSV-DS-Cav1-B, respectively.

Fig. 5 shows the results of a rapid fluorescent focus inhibition test (RFFIT test), which detects the ability of monoclonal antibodies to the RSV F protein to inhibit infection of Hep-2 cells by the RSV A2 strain.

Fig. 6 shows the results of the RFFIT test, which reveals the ability of monoclonal antibodies to the RSV F protein to inhibit infection of Hep-2 cells with RSV strain 18537.

Fig. 7 shows the results of inhibition of RSV infection of Hep-2 cells by monoclonal antibodies to the RSV F protein, isolated from clinical samples.

SEQUENCE DESCRIPTION

SEQ ID NO:1 represents the nucleotide sequence derived from Escherichia coli encoding the chloramphenicol resistance gene promoter (Cat promoter).

SEQ ID NO:2 is the nucleotide sequence of the Escherichia-targeting lambda phage encoding the attP integration site.

SEQ ID NO:3 is the nucleotide sequence derived from E. coli encoding the CDF origin of replication.

SEQ ID NO:4 is the nucleotide sequence derived from E. coli encoding the attB integration site.

SEQ ID NO:5 is the nucleotide sequence of E. coli derived chloramphenicol resistance gene.

SEQ ID NO:6 is the nucleotide sequence derived from E. coli encoding the expression element for the complete ampicillin resistance gene.

SEQ ID NO:7 is the nucleotide sequence of the Escherichia-targeting lambda phage encoding the inducible integrase expression element of the lambda phage lactose operon.

SEQ ID NO:8 represents the nucleotide sequence encoding the attL recombination site.

SEQ ID NO:9 represents the nucleotide sequence encoding the attR recombination site.

SEQ ID NO:10 represents the amino acid sequence of the F protein of RSV subtype A strain A2.

SEQ ID NO:11 represents the amino acid sequence of the F protein of RSV subtype B strain 18537.

SEQ ID NO:12 represents the amino acid sequence of the F protein of the RSV-DS-Cav1-A mutant structure with DS-Cav1 before fusion.

SEQ ID NO:13 represents the amino acid sequence of the F protein of the RSV-DS-Cav1-B mutant structure with DS-Cav1 before fusion.

SEQ ID NO:14 represents the amino acid sequence of the histidine tag.

SEQ ID NO:15 represents the amino acid sequence of the constant region of the heavy chain of human ( homo sapiens ) subtype IgG1.

SEQ ID NO:16 represents the amino acid sequence of the heavy chain constant region of the mutant human ( Homo sapiens ) IgG1-YTE subtype IgG1.

SEQ ID NO:17 represents the amino acid sequence of the constant region of the heavy chain of mouse ( Mus musculus ) subtype IgG2a.

SEQ ID NO:18 represents the amino acid sequence of the constant region of the light chain of the kappa subtype of human ( Homo sapiens ).

SEQ ID NO:19 represents the amino acid sequence of the light chain constant region of the lambda subtype of human ( Homo sapiens ).

SEQ ID NO:20 represents the amino acid sequence of the mouse ( Mus musculus ) kappa subtype light chain constant region.

SEQ ID NO:21 represents the amino acid sequence of the light chain constant region of the mouse ( Mus musculus ) lambda subtype.

SEQ ID NO:22 represents the amino acid sequence of the heavy chain variable region of the humanized monoclonal antibody to RSV palivizumab (Hu1129).

SEQ ID NO:23 represents the amino acid sequence of the light chain variable region of the humanized monoclonal antibody to RSV palivizumab (Hu1129).

SEQ ID NO:24 represents the amino acid sequence of the heavy chain variable region of the fully human monoclonal antibody to RSV nirsevimab (MEDI8897).

SEQ ID NO:25 represents the amino acid sequence of the light chain variable region of the fully human monoclonal antibody to RSV nirsevimab (MEDI8897).

SEQ ID NO:26 represents the amino acid sequence of the heavy chain variable region of the fully human monoclonal antibody to RSV clesrovimab (RB1).

SEQ ID NO:27 represents the amino acid sequence of the light chain variable region of the fully human monoclonal antibody to RSV clesrovimab (RB1).

SEQ ID NO:28 represents the amino acid sequence of the heavy chain variable region of the R3B1h1 Fab antibody to the RSV F protein.

SEQ ID NO:29 represents the amino acid sequence of the light chain variable region of the R3B1h1 Fab antibody to the RSV F protein.

SEQ ID NO:30 represents the amino acid sequence of the heavy chain variable region of the R22B1 Fab antibody to the RSV F protein.

SEQ ID NO:31 represents the amino acid sequence of the light chain variable region of the R22B1 Fab antibody to the RSV F protein.

SEQ ID NO:32-34 represent the amino acid sequences of HCDR1, HCDR2 and HCDR3 of the heavy chain variable region of the Fab antibody R3B1h1 to the RSV F protein, respectively.

SEQ ID NO:35-37 represent the amino acid sequences of LCDR1, LCDR2 and LCDR3 of the light chain variable region of the Fab antibody R3B1h1 to the RSV F protein, respectively.

SEQ ID NO:38-40 represent the amino acid sequences of HCDR1, HCDR2 and HCDR3 of the heavy chain variable regions of the Fab antibody R22B1 to the RSV F protein, respectively.

SEQ ID NO:35, 36,43 represent the amino acid sequences of LCDR1, LCDR2 and LCDR3 of the light chain variable regions of the Fab antibody R22B1 to the RSV F protein, respectively.

SEQ ID NO:44 is the nucleotide sequence of E. coli encoding the pBR origin of replication.

SEQ ID NO:45 is the nucleotide sequence of the f1 phage encoding the f1 origin of replication.

SEQ ID NO:46 is the nucleotide sequence of phage M13 encoding the gIII protein of filamentous phage M13.

IMPLEMENTATION OF THE INVENTION

Integration of specific recombinants and cleavage of the target nucleotide sequence in vivo and in vitro can be achieved by genetic engineering using a specific recombinant system or a recombinant cell with phage lambda integrase (lambda integrase). According to the literature, specific recombination of plasmids for expressing the antibody light chain and plasmids for expressing the antibody heavy chain in cells can be achieved using genetically engineered bacteria expressing lambda integrase through heat induction to produce a complete plasmid expressing a functional Fab fragment ¹⁰ . Using Thermo Fisher's Gateway homologous recombination technology, which utilizes the specific recombination principle of phage lambda integrase, in vitro recombination efficiency can reach 95% without the need for traditional gene cloning steps such as digestion and ligation. This technology is used to create the Gateway kit for constructing a cDNA library with 1.0E+7 diversity.

The invention according to the present application makes it possible to overcome the disadvantage of insufficient efficiency of DNA transformation when constructing an antibody library by means of efficient introduction of phage using a recombinant system or a recombinant cell with phage lambda integrase. The introduction of, in particular, the VH-CH1 heavy chain expression element makes it possible to induce E. coli with plasmids expressing lambda integrase and containing the antibody light chain expression element, and to obtain a large-size Fab antibody library (more than 1.0E+12) by specific recombination of the heavy chain expression element (e.g., the VH-CH1 heavy chain expression element) of the antibody and the light chain expression element of the antibody using lambda integrase in vivo , and to perform screening for antibiotic resistance using phage with the heavy chain expression element (e.g., the VH-CH1 heavy chain expression element) of the antibody packaged therein for the introduction of E. coli capable of inducing the expression of lambda integrase and containing a plasmid containing the antibody light chain expression element.

The inventors of the present invention utilized the constructed Fab antibody library discussed above to screen for novel RSV Fab antibodies. Various aspects of the present invention provide a novel RSV antibody, a nucleic acid molecule encoding the antibody, a vector comprising such a nucleic acid molecule, a host cell comprising such a nucleic acid molecule or such a vector, a composition comprising the antibody, a method for producing and purifying the antibody, and medical and biological uses for such an antibody. Based on the amino acid sequences of the variable regions of the antibody provided in the present application, a full-length antibody molecule can be produced as a medicament for the prevention or treatment of RSV-related diseases.

Except where otherwise stated, the inventions of this application may be implemented using techniques known in the fields of molecular biology, microbiology, cytology, biochemistry and immunology.

Except where otherwise noted, the terms used in this application have their commonly understood meanings as understood by those skilled in the art.

Definitions

As used herein, the term "recombination site" means a site for phage integration in the phage genome and the bacterial genome. In some embodiments of the present invention, the first recombination site and the second recombination site may be a site for phage integration in the phage and host bacterium genomes, respectively, including when the first recombination site is a site for phage integration in the phage genome, while the second recombination site is a site for phage integration in the genome of the phage host bacteria; or when the second recombination site is a site for phage integration in the phage genome, and the first recombination site is a site for phage integration in the genome of the phage host bacterium.

As used herein, the term "plasmid" refers to a DNA molecule other than a chromosome (or nucleoid) in an organism such as bacteria, yeast, actinomycetes, etc. It is a circular, double-stranded DNA molecule located in the cytoplasm (except for yeast, where the 2-micrometer yeast plasmid is located in the nucleus), capable of autonomous replication such that a constant number of copies is also maintained in the progeny, and expressing the genetic information it carries. As used herein, the term "plasmid" includes the meanings of "bacterial plasmid,""phagemid," etc. A "bacterial plasmid" is a vector widely used in DNA recombination techniques, as well as a vehicle for delivering a useful exogenous gene into a recipient cell for proliferation and expression by genetic engineering means. A "phagemid" is a vector formed by combining a plasmid vector with a single-stranded phage vector, containing a single-stranded phage packaging sequence, a replicon, a plasmid replicon, a cloning site, a marker gene, etc. In host cells, particularly E. coli , it is capable of replication in the same way as conventional double-stranded plasmid DNA molecules, and, in the presence of helper phages, it is capable of replication according to the "rolling circle" model, forming single-stranded DNA that can be packaged into a phage particle.

In this document, the term "attP integration site" means a specific sequence in phage DNA with a 15 base pair (bp) core region with a sequence identical to that of the host chromosome.

In this document, the term "attB integration site" means an integration site in the bacterial genome with a 15 bp core region with a sequence identical to the corresponding phage gene sequence.

In this document, the term "phage integrase" means a protein with type 1 topoisomerase activity that specifically binds to the attP site and the attB integration site and is essential for the integration of the phage into the host genome by site-specific recombination.

As used herein, the term "origin of replication" means the site of origin of plasmid replication, consisting of the origin replicon (ORI) and the corresponding regulatory element.

As used herein, the term "antibody" refers to an immunoglobulin molecule capable of specifically binding to a target via at least one antigen recognition site in the variable region of the immunoglobulin molecule. Targets include, but are not limited to, carbohydrates, polynucleotides, lipids, polypeptides, and the like. In the context of this document, the term "antibody" includes not only the meaning of "intact (i.e., full-length) antibody", but also its antigen-binding fragment (e.g., Fab fragment, Fab' fragment, F(ab') ₂ fragment or Fv fragment), its variant, fusion protein containing parts of the antibody, humanized antibody, chimeric antibody, diabody, linear antibody, single-chain variable fragment, multispecific antibody (e.g., bispecific antibody), as well as any modified configurations of the immunoglobulin molecule containing the desired recognition site of a specific antigen, including a glycosylated variant of the antibody, an amino acid sequence variant of the antibody and a covalently modified antibody.

An intact or full-length antibody typically contains two heavy chains and two light chains. Each heavy chain contains a heavy chain variable region (VH region) and the first, second, and third constant regions (CH1, CH2, and CH3). Each light chain contains a light chain variable region (VL region) and a constant region (CL region). A full-length antibody can be any type, specifically an IgD, IgE, IgG, IgA, or IgM antibody (or a subtype thereof), but may not belong to any specific type. Immunoglobulins are classified into one type or another depending on the amino acid sequences of the constant domains of their heavy chains. Generally, immunoglobulins are classified into five main types, i.e. IgA, IgD, IgE, IgG, and IgM, some of which can be further subdivided into subtypes (isotypes), such as IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The constant domains of the heavy chains associated with individual immunoglobulin types are designated α, δ, ε, γ, and μ, respectively. The subunits and three-dimensional structures of the various immunoglobulin types are well known.

As used herein, the term "antigen-binding fragment" or "antigen-binding portion" refers to the portion or region of an intact antibody molecule responsible for antigen binding. An antigen-binding domain may comprise the variable region of the heavy chain (VH region), the variable region of the light chain (VL region), or both. Both the VH and VL regions typically contain three complementarity-determining regions, i.e., CDR1, CDR2, and CDR3.

It is well known to those skilled in the art that complementarity-determining regions (CDRs, typically CDR1, CDR2, and CDR3) are the regions of the variable region that most influence the affinity and specificity of an antibody. The amino acid sequences of the CDRs of the VH or VL region are determined using two generally accepted nomenclatures: the Chothia nomenclature and the Kabot nomenclature ^11-13 . For the amino acid sequences of the variable regions of an antibody, the amino acid sequences of the CDRs in the VH region and the amino acid sequences of the VL region can be determined using the Chothia nomenclature or the Kabot nomenclature. In one embodiment of the present invention, the amino acid sequences of the CDRs are determined using the Kabot nomenclature.

In the case of amino acid sequences of the variable regions of a particular antibody, the amino acid sequences of the CDR regions in the amino acid sequences of the variable regions can be determined by various methods, for example, using the Abysis online program (http://www.abysis.org/).

Examples of antigen-binding fragments include, but are not limited to: (1) a Fab fragment, which may be a monovalent fragment of an antibody with a VL-CL type chain and a VH-CH1 type chain; (2) an F(ab') ₂ fragment, which may be a bivalent fragment with two Fab' fragments linked by a hinge disulfide bridge (i.e., a Fab'dimer); (3) an Fv fragment with both VL and VH domains in one arm of an antibody; (4) a single-chain Fv fragment (scFv fragment), which may be a single polypeptide chain consisting of a VH domain and a VL domain linked by a polypeptide linker; and (5) (scFv fragment) ₂ , which may contain two VH domains linked by a peptide linker and two VL domains connected to two VH domains by a disulfide bridge.

In some particular embodiments of the present invention, the antibody is a Fab fragment, i.e. a monovalent fragment with a VL-CL type chain and a VH-CH1 type chain.

In this document, the term "Fd fragment" means a fragment consisting of the variable region of the heavy chain (VH region) of an antibody and the first constant region (CH1) of the heavy chain of the antibody.

As used herein, the term "specific binding" means a non-random binding reaction between two molecules, such as the binding of an antibody to an antigenic epitope.

As used herein, the term "neutralizing antibody" means an antibody that is capable of binding to an antigen on the surface of a pathogenic microorganism, thereby inhibiting cell invasion by preventing the attachment of the pathogenic microorganism to a target cell receptor or by preventing the fusion of the viral envelope with the cell membrane.

As used herein, the term "monoclonal antibody" means an antibody obtained from a substantially homogeneous population of antibodies, i.e., from a population in which the antibodies comprising it are identical except for naturally occurring mutations that may be present in a small number of individuals.

According to a first aspect, the present application provides an antibody to respiratory syncytial virus (RSV) comprising a heavy chain variable region comprising HCDR1, HCDR2 and HCDR3 and a light chain variable region comprising LCDR1, LCDR2 and LCDR3, wherein

the amino acid sequence of HCDR1 is presented in SEQ ID NO:32, the amino acid sequence of HCDR2 is presented in SEQ ID NO:33, the amino acid sequence of HCDR3 is presented in SEQ ID NO:34, the amino acid sequence of LCDR1 is presented in SEQ ID NO:35, the amino acid sequence of LCDR2 is presented in SEQ ID NO:36, and the amino acid sequence of LCDR3 is presented in SEQ ID NO:37; or

the amino acid sequence of HCDR1 is presented in SEQ ID NO:38, the amino acid sequence of HCDR2 is presented in SEQ ID NO:39, the amino acid sequence of HCDR3 is presented in SEQ ID NO:40, the amino acid sequence of LCDR1 is presented in SEQ ID NO:35, the amino acid sequence of LCDR2 is presented in SEQ ID NO:36, and the amino acid sequence of LCDR3 is presented in SEQ ID NO:43;

wherein the amino acid sequences of the complementarity-determining regions of the heavy chain and the complementarity-determining regions of the light chain are determined according to the Cabot nomenclature.

In some embodiments of the first aspect, the amino acid sequence of the heavy chain variable region of the antibody is shown in SEQ ID NO: 28 or 30.

In some embodiments of the first aspect, the amino acid sequence of the light chain variable region of the antibody is shown in SEQ ID NO: 29 or 31.

In some embodiments of the first aspect, the amino acid sequence of the heavy chain variable region of the antibody is shown in SEQ ID NO: 28, wherein the amino acid sequence of the light chain variable region of the antibody is shown in SEQ ID NO: 29; or

the amino acid sequence of the variable region of the heavy chain of the antibody is presented in SEQ ID NO: 30, while the amino acid sequence of the variable region of the light chain of the antibody is presented in SEQ ID NO: 31.

According to a second aspect, the present application provides an antibody to respiratory syncytial virus (RSV), wherein the amino acid sequence of the heavy chain variable region of the antibody is at least 90% identical to the amino acid sequence defined in SEQ ID NO: 28 or 30, while the amino acid sequence of the light chain variable region of the antibody is at least 90% identical to the amino acid sequence defined in SEQ ID NO: 29 or 31.

In some embodiments of the second aspect, the amino acid sequence of the heavy chain variable region of the antibody is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical to the amino acid sequence defined in SEQ ID NO: 28 or 30.

In some embodiments of the second aspect, the amino acid sequence of the light chain variable region of the antibody is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical to the amino acid sequence defined in SEQ ID NO: 29 or 31.

In some embodiments of the second aspect, the amino acid sequence of the heavy chain variable region of the antibody differs from the amino acid sequence defined in any of SEQ ID NOs: 28 and 30 by about 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acids that are replaced, deleted and/or added.

In some embodiments of the second aspect, the amino acid sequence of the light chain variable region of the antibody differs from the amino acid sequence defined in any of SEQ ID NOs: 29 and 31 by about 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acids substituted, deleted and/or added.

In some embodiments of the second aspect, the C-terminal or N-terminal region of the amino acid sequence presented in any of SEQ ID NOs: 28 and 30 may also be shortened by about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25 or more amino acids, wherein the resulting amino acid sequences retain a function similar to that of the variable region of the heavy chain of an antibody.

In some embodiments of the second aspect, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25 or more amino acids may also be added to the C-terminal or N-terminal region of the amino acid sequence presented in any of SEQ ID NOs: 28 and 30, wherein the resulting amino acid sequences retain a function similar to that of the variable region of the heavy chain of an antibody.

In some embodiments of the second aspect, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25 or more amino acids may also be added or deleted in a region that is not the C-terminus or N-terminus of the amino acid sequence presented in any of SEQ ID NOs: 28 and 30, provided that the altered amino acid sequences substantially retain a function similar to that of the heavy chain variable region of an antibody.

In some embodiments of the second aspect, the C-terminal or N-terminal region of the amino acid sequence presented in any of SEQ ID NOs: 29 and 31 may also be shortened by about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25 or more amino acids, wherein the resulting amino acid sequences retain a function similar to that of the variable region of the light chain of an antibody.

In some embodiments of the second aspect, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25 or more amino acids may also be added to the C-terminal or N-terminal region of the amino acid sequence presented in any of SEQ ID NOs: 29 and 31, wherein the resulting amino acid sequences retain a function similar to that of the variable region of the light chain of an antibody.

In some embodiments of the second aspect, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25 or more amino acids may also be added or deleted in a region that is not the C-terminus or N-terminus of the amino acid sequence presented in any of SEQ ID NOs: 29 and 31, provided that the altered amino acid sequences substantially retain a function similar to that of the variable region of the light chain of an antibody.

In some embodiments of the first or second aspects, the antibody is a neutralizing antibody.

In some embodiments of the first or second aspects, the antibody may be capable of binding to the F protein of human respiratory syncytial virus (RSV).

In some particular embodiments of the first or second aspects, the human respiratory syncytial virus (RSV) F protein is a recombinant human RSV F protein, in particular a recombinant human RSV F protein as defined in SEQ ID NO: 12 or 13.

In some embodiments of the first or second aspects, the antibody is a Fab fragment, a whole antibody, an F(ab') ₂ fragment, or a single-chain Fv fragment (scFv fragment).

In some particular embodiments of the first or second aspects, the antibody is a Fab fragment.

In some embodiments of the first or second aspects, the antibody is a monoclonal antibody.

In some embodiments of the first or second aspects, the antibody comprises a heavy chain constant region of an IgG1 subtype, an IgG2 subtype, or an IgG4 subtype.

In some embodiments of the first or second aspects, the heavy chain constant region comprises an Fc region of an IgG1 subtype heavy chain constant region, wherein the amino acids at positions 252, 254, 256 of the Fc region sequence are Y, T and E, respectively, wherein the amino acid positions of the antibody constant region are determined according to EU numbering.

In some embodiments of the first or second aspects, the antibody comprises a light chain constant region of the kappa subtype or the lambda subtype.

According to a third aspect, the present application provides a nucleic acid molecule encoding an antibody according to the first or second aspect.

In some embodiments of the third aspect, the nucleic acid molecule is operably linked to a regulatory nucleic acid sequence that can be recognized by a host cell transformed with the vector.

According to a fourth aspect, the present application provides a pharmaceutical composition comprising an antibody according to the first or second aspect and a pharmaceutically acceptable excipient, diluent or carrier.

In some embodiments of the fourth aspect, the pharmaceutical composition is for the prevention or treatment of an RSV-induced disease.

In some embodiments of the fourth aspect, the RSV-induced disease is an infectious respiratory disease, in particular bronchiolitis and pneumonia.

In some embodiments of the fourth aspect, the pharmaceutical composition may further comprise one or more of the following: a lubricant, in particular talc, magnesium stearate and mineral oil; a wetting agent; an emulsifier; a suspending agent; a preservative, in particular benzoic acid, sorbic acid and calcium propionate; a sweetener and/or flavoring agent, and the like.

In some embodiments of the fourth aspect, the pharmaceutical compositions of the present application may be formulated as a tablet, dragee, powder, lozenge, elixir, suspension, emulsion, solution, syrup, suppository, or capsule, etc.

In some embodiments of the fourth aspect, the pharmaceutical compositions of the present application can be administered by any physiologically acceptable route, including, but not limited to, orally, parenterally, intranasally, rectally, intraperitoneally, by intravascular injection, subcutaneously, transdermally, by inhalation, etc.

In some embodiments of the fourth aspect, the pharmaceutical composition for therapeutic use may be formulated into a storable dosage form as a lyophilized formulation or an aqueous solution by mixing a reagent of the desired purity with an appropriate pharmaceutically acceptable carrier, excipient, etc.

According to a fifth aspect, the present application provides the use of an antibody according to the first or second aspect, a nucleic acid molecule according to the third aspect, or a pharmaceutical composition according to the fourth aspect in the manufacture of a medicament for the prevention or treatment of RSV-induced diseases.

In some embodiments of the fifth aspect, the RSV-induced disease is an infectious respiratory disease, in particular bronchiolitis and pneumonia.

According to a sixth aspect, the present application provides a method for preventing or treating a disease caused by RSV, comprising the step of administering to a subject in need thereof an antibody according to the first or second aspect or a pharmaceutical composition according to the fourth aspect.

In some embodiments of the sixth aspect, the RSV-induced disease is an infectious respiratory disease, in particular bronchiolitis and pneumonia.

According to a seventh aspect, the present application provides a combination of plasmids for expressing a library of Fab fragments of antibodies, comprising a first plasmid and a second plasmid, wherein

the first plasmid contains the VH-CH1 expression element of the antibody heavy chain and the first recombination site;

the second plasmid contains the VH-CH1 expression element of the antibody light chain and the second recombination site; wherein

the first recombination site is different from the second recombination site.

In some embodiments of the seventh aspect, the combination of plasmids comprises a plurality of first plasmids, wherein the plurality of first plasmids carry different VH-CH1 antibody heavy chain expression elements and have an identical first recombination site.

In some embodiments of the seventh aspect, the plasmid combination comprises a plurality of second plasmids, wherein the plurality of second plasmids carry different VH-CH1 antibody light chain expression elements and have an identical second recombination site.

According to an eighth aspect, the present application provides a recombinant system for expressing a library of Fab fragments of antibodies, comprising a first plasmid, a second plasmid and a phage integrase-expressing cell; wherein

the first plasmid contains the VH-CH1 expression element of the antibody heavy chain and the first recombination site;

the second plasmid contains the VH-CH1 expression element of the antibody light chain and the second recombination site; wherein

the first recombination site is different from the second recombination site.

In some embodiments of the eighth aspect, the recombinant system comprises a plurality of first plasmids, wherein the plurality of first plasmids carry different VH-CH1 antibody heavy chain expression elements and have an identical first recombination site.

In some embodiments of the eighth aspect, the recombinant system comprises a plurality of second plasmids, wherein the plurality of second plasmids carry different VH-CH1 light chain antibody expression elements and have an identical second recombination site.

In some embodiments of the eighth aspect, the first plasmid, the second plasmid and the phage integrase-expressing cell exist independently of one another, or at least one of the first plasmid and the second plasmid has been introduced into the phage integrase-expressing cell.

In some particular embodiments of the eighth aspect, the second plasmid is introduced into a phage integrase-expressing cell.

According to a ninth aspect, the present application provides a recombinant cell for expressing a library of Fab fragments of antibodies, comprising a first plasmid, a second plasmid and expressing a phage integrase; wherein

the first plasmid contains the VH-CH1 expression element of the antibody heavy chain and the first recombination site;

the second plasmid contains the VH-CH1 expression element of the antibody light chain and the second recombination site; wherein

the first recombination site is different from the second recombination site.

In some embodiments of the ninth aspect, the recombinant cell comprises a plurality of first plasmids, wherein the plurality of first plasmids carry different VH-CH1 antibody heavy chain expression elements and have an identical first recombination site.

In some embodiments of the ninth aspect, the recombinant cell comprises a plurality of second plasmids, wherein the plurality of second plasmids carry different VH-CH1 light chain antibody expression elements and have an identical second recombination site.

In some embodiments of the ninth aspect, the recombinant cell is a prokaryotic cell.

In some embodiments of the ninth aspect, the recombinant cell is E. coli .

In some embodiments of the ninth aspect, the recombinant cell is a genetically engineered bacterium.

In some particular embodiments of the ninth aspect, the recombinant cell is a male E. coli strain with a sex factor (F-factor), in particular the TG1 strain.

According to a tenth aspect, the present application provides a method for producing a library of Fab fragments of antibodies, which comprises transferring a first plasmid and a second plasmid into a cell expressing phage integrase; wherein

the first plasmid contains the VH-CH1 expression element of the antibody heavy chain and the first recombination site;

the second plasmid contains the VH-CH1 expression element of the antibody light chain and the second recombination site; wherein

the first recombination site is different from the second recombination site.

In some embodiments of the tenth aspect, a plurality of first plasmids are transferred into a phage integrase-expressing cell, wherein the plurality of first plasmids carry different VH-CH1 antibody heavy chain expression elements and have an identical first recombination site.

In some embodiments of the tenth aspect, a plurality of second plasmids are transferred into a phage integrase-expressing cell, wherein the plurality of second plasmids carry different VH-CH1 antibody light chain expression elements and have an identical second recombination site.

In some embodiments of the tenth aspect, transferring the first plasmid and/or the second plasmid into the phage integrase-expressing cell comprises the steps of:

(1) transfer the first plasmid and/or the second plasmid into a competent cell;

(2) infecting the competent cell obtained in step (1) with a helper phage to construct a phage library; and

(3) transfer the phage library obtained in step (2) into a cell expressing phage integrase.

In some embodiments of the tenth aspect, the competent cell is a prokaryotic cell.

In some embodiments of the tenth aspect, the competent cell is E. coli .

In some embodiments of the tenth aspect, the competent cell is a genetically engineered bacterium.

In some particular embodiments of the tenth aspect, the competent cell is a male E. coli strain with an F factor, in particular the TG1 strain.

In some embodiments of the tenth aspect, the competent cell does not express phage integrase.

In some embodiments of the tenth aspect, the helper phage is an M13 helper phage.

In some embodiments of any of the seventh to tenth aspects, the first plasmid is a phagemid.

In some embodiments of any of the seventh to tenth aspects, the second plasmid is a phagemid.

In some embodiments of any of the seventh to tenth aspects, the first plasmid and the second plasmid have different origins of replication.

In some embodiments of any of the seventh to tenth aspects, the origin of replication is selected from the group consisting of one or more of the pBR origin of replication, the CDF origin of replication, the M13 filamentous phage origin of replication (f1 origin of replication), and the p15A origin of replication.

In some particular embodiments of any of the seventh to tenth aspects, the first plasmid comprises a pBR origin of replication and an f1 origin of replication.

In some particular embodiments of any of the seventh to tenth aspects, the second plasmid comprises a CDF origin of replication.

In some embodiments of any of the seventh to tenth aspects, the first recombination site and the second recombination site are phage integration sites in the genome of the phage and the host bacterium, respectively.

In some embodiments of any of the seventh to tenth aspects, the first recombination site is an attP site.

In some embodiments of any of the seventh to tenth aspects, the second recombination site is an attB site.

In some particular embodiments of any of the seventh to tenth aspects, the first recombination site comprises a nucleotide sequence defined in SEQ ID NO:2, or a nucleotide sequence derived from the nucleotide sequence defined in SEQ ID NO:2 by one or more substitutions, deletions or additions that do not substantially alter the function of the first recombination site.

In some particular embodiments of any of the seventh to tenth aspects, the second recombination site comprises a nucleotide sequence defined in SEQ ID NO:4, or a nucleotide sequence derived from the nucleotide sequence defined in SEQ ID NO:4 by one or more substitutions, deletions or additions that do not substantially alter the function of the second recombination site.

In some embodiments of any of the seventh to tenth aspects, the number of one or more substitutions, deletions or additions that do not substantially alter the function of the first recombination site or the second recombination site is from 1 to 30, preferably from 1 to 20, more preferably from 1 to 10, wherein the function of the resulting nucleotide sequence remains substantially unchanged compared to the function of the first recombination site or the second recombination site.

In some embodiments of any of the seventh to tenth aspects, the first plasmid and the second plasmid are different from each other.

In some particular embodiments of any of the seventh to tenth aspects, the first plasmid is a phagemid, wherein the second plasmid is a prokaryotic expression vector (e.g., a bacterial plasmid).

In some particular embodiments of any of the seventh to tenth aspects, the first plasmid is pHGDisn-attP-site-new.

In some particular embodiments of any of the seventh to tenth aspects, the second plasmid is pHKb-attB-site-new.

In some embodiments of any of the seventh to tenth aspects, the first recombination site is located at a polyclonal enzyme cleavage site of the first plasmid.

In some embodiments of any of the seventh to tenth aspects, the second recombination site is located at a polyclonal enzyme cleavage site of the second plasmid.

In some particular embodiments of any of the seventh to tenth aspects, the first recombination site is located between the XbaI and KpnI enzymatic cleavage sites of the pHGDisn-attP-site-new plasmid.

In some particular embodiments of any of the seventh to tenth aspects, the second recombination site is located between the HindIII and BamHI enzymatic cleavage sites of the pHKb-attB-site-new plasmid.

In some embodiments of any of the seventh to tenth aspects, the light chain constant region (CL region) of the antibody is a human kappa light chain constant region or a human lambda light chain constant region.

In some embodiments of any of the seventh to tenth aspects, the CH1 domain of the heavy chain constant region of the antibody is of the IgG1, IgG2, IgG3 or IgG4 subtype.

In some embodiments of any of the seventh to tenth aspects, the first plasmid and/or the second plasmid comprises a resistance gene encoding region.

In some embodiments of any of the seventh to tenth aspects, a nucleic acid molecule comprising a nucleic acid molecule encoding the CH1 domain of the constant region of an antibody heavy chain is fused at its 3' end to the 5' end of a nucleic acid molecule encoding the gIII protein of the filamentous phage M13.

In some embodiments of any of the seventh to tenth aspects, the presence of the resistance gene facilitates screening of the recombinant system or recombinant cell.

In some embodiments of any of the seventh to tenth aspects, the resistance gene is an antibiotic resistance gene.

In some embodiments of any of the seventh to tenth aspects, the resistance gene is selected from the group consisting of: a chloramphenicol resistance gene (CmR gene), an ampicillin resistance gene (Ampr gene), a kanamycin resistance gene (KaR gene), and a tetracycline resistance gene (TetR gene).

In some embodiments of any of the eighth to tenth aspects, the phage integrase is tyrosine integrase.

In some embodiments of any of the eighth to tenth aspects, the tyrosine integrase is phage lambda integrase.

In some embodiments of any of the eighth to tenth aspects, the phage integrase is an inducibly expressed or constitutively expressed integrase.

In some embodiments of any of the eighth to tenth aspects, the phage integrase is an inducibly expressed integrase.

In some embodiments of any of the eighth to tenth aspects, inducible expression is carried out using an inducible promoter, in particular a lactose promoter (Plac) or an arabinose promoter (Para).

In some embodiments of the eighth or tenth aspect, the phage integrase-expressing cell is a prokaryotic cell.

In some embodiments of the eighth or tenth aspect, the phage integrase-expressing cell is E. coli .

In some embodiments of the eighth or tenth aspect, the phage integrase-expressing cell is a genetically engineered bacterium.

In some embodiments of the eighth or tenth aspect, the phage integrase-expressing cell is a male E. coli strain with an F factor.

In some particular embodiments of the eighth or tenth aspect, the phage integrase-expressing cell is strain TG1.

In some particular embodiments of any of the seventh to tenth aspects, the first plasmid comprises an antibody heavy chain VH-CH1 expression element and an attP integration site; wherein the second plasmid comprises an antibody light chain VH-CH1 expression element and an attB integration site.

In some embodiments of the seventh aspect, the first recombination site and the second recombination site correspond to each other. For example, if the nucleotide sequence of the first recombination site is presented in SEQ ID NO:2, then the nucleotide sequence of the second recombination site is presented in SEQ ID NO:4.

In some embodiments of the eighth or tenth aspect, there is a correspondence between the first recombination site, the second recombination site, the type of phage integrase, and the type of cell expressing the phage integrase.

In some embodiments of the eighth or tenth aspect, the second recombination site, the type of phage integrase and the type of cell expressing the phage integrase useful in the invention of the present application may be determined once the first recombination site is determined.

In some embodiments of the eighth or tenth aspect, the first recombination site, the type of phage integrase and the type of cell expressing the phage integrase useful in the invention of the present application may be determined once the second recombination site is determined.

In some embodiments of the eighth or tenth aspect, in the case of determining the type of phage integrase, the type of cell expressing the phage integrase useful in the invention of the present application can be determined once the type of phage integrase is determined.

In some embodiments of the eighth or tenth aspect, if the nucleotide sequence of the first recombination site is presented in SEQ ID NO:2, and the nucleotide sequence of the second recombination site is presented in SEQ ID NO:4, then the phage integrase is phage lambda integrase, and the phage integrase-expressing cell is E. coli strain TG1.

In some embodiments of the ninth aspect, there is a correspondence between the first recombination site, the second recombination site, the phage integrase type, and the recombinant cell type.

In some embodiments of the ninth aspect, the second recombination site, the phage integrase type and the recombinant cell type useful in the invention of the present application may be determined once the first recombination site is determined.

In some embodiments of the ninth aspect, the first recombination site, the phage integrase type and the recombinant cell type useful in the invention of the present application may be determined once the second recombination site is determined.

In some embodiments of the ninth aspect, the type of recombinant cell useful in the invention of the present application may be determined once the type of phage integrase is determined.

In some embodiments of the ninth aspect, if the nucleotide sequence of the first recombination site is presented in SEQ ID NO:2, and the nucleotide sequence of the second recombination site is presented in SEQ ID NO:4, then the phage integrase is phage lambda integrase, wherein the recombinant cell is E. coli strain TG1.

In some particular embodiments of the tenth aspect, the method for producing a library of Fab fragments of antibodies may include the steps of:

(1) transferring a first plasmid (e.g., a plurality of first plasmids, in particular, a plurality of phagemids carrying different VH-CH1 heavy chain antibody expression elements and an identical attP integration site) into competent cells (e.g., competent cells of the TG1 strain);

(2) infecting the competent cells obtained in step (1) with a helper phage (e.g., an M13 helper phage) to construct a phage library (e.g., a phage library comprising multiple phages carrying different VH-CH1 expression elements of the heavy chain of antibodies and an identical attP integration site);

(3) transfer a second plasmid (e.g., multiple second plasmids carrying different VH-CH1 light chain antibody expression elements and an identical attB integration site) into cells expressing phage integrase (e.g., competent cells of the TG1 strain expressing phage lambda integrase);

(4) induce the expression of a phage integrase (e.g., lambda phage integrase) by the cells obtained in step (3);

(5) infecting the phage integrase-expressing cells obtained in step (4) with a phage library (e.g., a phage library comprising multiple phages carrying different VH-CH1 expression elements of the antibody heavy chain and an identical attP integration site); and

(6) infect the cells obtained in step (5) with a helper phage (e.g., M13 helper phage) to construct a library of Fab fragments of antibodies.

According to an eleventh aspect, the present application provides the use of a combination of plasmids according to the seventh aspect, a recombinant system according to the eighth aspect, or a recombinant cell according to the ninth aspect in the production of a library of Fab fragments of antibodies.

In other aspects, the present application provides a vector comprising a nucleic acid molecule encoding an antibody of the present application, or a light or heavy chain thereof, a host cell comprising such a vector, and a method for producing such an antibody. In some embodiments, the nucleic acid molecule is operably linked to a regulatory sequence that can be recognized by host cells transformed with the vector. In some embodiments, the method for producing an antibody comprises culturing the host cells. In some embodiments, the method for producing an antibody further comprises isolating the antibody from the culture medium of the host cells.

Furthermore, the RSV-specific antibody disclosed herein can be used to detect RSV in a biological sample. Antibody-based detection methods are well known in the art and include, for example, ELISA, immunoblotting, radioimmunoassay, immunofluorescence, immune precipitation, and other related techniques.

It should be understood that the above detailed description is intended solely to provide those skilled in the art with a clearer understanding of the invention of this application, but is not intended to limit it in any respect. Those skilled in the art will be able to make various modifications and variations to the embodiments disclosed above.

Examples

The following examples are for illustrative purposes only and are not intended to limit the scope of the invention according to this application.

Example 1: Generation of a library of fully human recombinant Fab fragments

This example was implemented based on Chinese Patent for Invention No. CN 108251431B, the contents of which are incorporated into this application by reference.

1.1 Construction of recombinant expression plasmids containing the attB site and the attP integration site

Based on the pADG-S plasmid, a pHGDisn-attP-site-new phagemid vector containing the attP integration site and expressing the Fd domain of the heavy chain (VH-CH1) of the antibody was constructed by cloning a fragment of a synthetic recombinant gene containing the promoter of the chloramphenicol resistance gene (Cat promoter, with the coding sequence defined in SEQ ID NO: 1) and the attP integration site (attP1, with the coding sequence defined in SEQ ID NO: 2) into the pADG-S vector using digestion with two enzymes, XbaI and KpnI (Fig. 1).

Based on the pADK-S plasmid, a prokaryotic expression vector pHKb-attB-site-new containing the attB integration site and expressing the antibody light chain was constructed by cloning a fragment of a synthetic recombinant gene containing the CDF origin of replication (CDF ori, with the coding sequence defined in SEQ ID NO: 3), the attB integration site (attB1, with the coding sequence defined in SEQ ID NO: 4), the chloramphenicol resistance gene (CmR gene, with the coding sequence defined in SEQ ID NO: 5) and the expression element for the complete ampicillin resistance gene (AmpR gene, with the coding sequence defined in SEQ ID NO: 6) into the pADK-S vector using digestion with two enzymes, HindIII and BamHI (Fig. 2).

pHGDisn-attP-site-new and pHKb-attB-site-new have different origins of replication and are capable of replication and coexistence within the same E. coli cell. Following induction of specific recombination of the attP site and attB integration site by phage lambda integrase (λInt), the newly formed plasmid can express chloramphenicol acetyltransferase and exhibit chloramphenicol resistance, as it contains the promoter and coding gene for the complete chloramphenicol resistance gene, thereby facilitating the screening of recombinant plasmids.

1.2 Obtaining the TG1 E. coli strain with an edited λInt genome

The attL site and attR site were created by specific recombination of the attB site and the attP integration site in E. coli cells, with the reaction required to be carried out under conditions of catalysis by phage lambda integrase ^14. The gene encoding the element expressing the inducible integrase of the phage lambda lactose operon (with the nucleotide sequence defined in SEQ ID NO: 7) was introduced into the genome of the E. coli strain TG1 using genome editing tools. Recombinant genetically engineered bacteria of the TG1-λInt strain were produced by GenScript Biotech Corporation.

1.3 Construction of an antibody library containing fully human heavy chains

A fully human heavy chain antibody library with a diversity of 1.73E+9 was constructed based on Chinese Invention Patent No. CN 108251431B by using frozen cDNAs of the native human heavy chain variable region and PCR products of VH1, VH3, and VH5 with mutations as templates, designing PCR primers to amplify fragments of interest, cloning the fragments of interest into the pHGDisn-attP-site-new vector by using dual enzyme digestion with NcoI and PmlI, and electrotransformation to introduce the ligation products into competent cells of the TG1 strain. The constructed antibody library was sequenced and analyzed with an average accuracy above 85%.

1.4 Construction of an antibody library containing fully human light chains

A fully human light chain antibody library with a diversity of 1.95E+8 was constructed based on Chinese Invention Patent No. CN 108251431B by using frozen cDNAs of the natural human variable light chain region and PCR products of VK1 and VK3 with mutations as templates, designing PCR primers to amplify fragments of interest, cloning the fragments of interest into the pHKb-attB-site-new vector by using dual enzyme digestion with NcoI and PmlI, and electrotransformation to introduce the ligation products into competent cells of the TG1-λInt strain. The constructed antibody library was sequenced and analyzed with an average accuracy above 90%.

1.5 Obtaining a library of fully human recombinant Fab fragments

A bacterial solution containing the heavy chain library constructed as described above was inoculated into 200 ml of liquid medium, grown to logarithmic growth phase at 37°C and 220 rpm, infected with M13 helper phage, and grown overnight at 28°C and 220 rpm for phage amplification. A purified heavy chain phage library was then obtained by PEG/NaCl precipitation. The purified heavy chain phage library, after titration, was stored in a refrigerator at -80°C as a stock.

The antibody library containing fully human light chains, prepared as described above, was inoculated into 50 ml of liquid medium containing IPTG at a final concentration of 0.1 mmol, grown to logarithmic growth phase at 37°C and 220 rpm, and then infected with the heavy chain phage library. After infection, the bacterial fluid was evenly distributed over a bacterial culture dish and incubated overnight in an incubator at 32°C.

Bacteria grown on the plate were enumerated and sent for sequencing to identify recombinant sites of monoclonals, as well as light and heavy chain sequences. The library size after recombination was estimated by the dilution method. According to the enumeration results, the total number of recombinant colonies was 3.80E+12. The results of monoclonal antibody sequencing showed that all clones grown on the plate were recombinant plasmids containing the newly formed attL site (attL1, with the coding sequence defined in SEQ ID NO: 8) and attR site (attR1, with the coding sequence defined in SEQ ID NO: 9) recombination, complete light chain genes and the Fd fragment of the heavy chain with a content of correct antibody genes greater than 76.5%.

Colonies on the above-mentioned bacterial culture plate were picked, inoculated into liquid medium, and cultured at 37°C and 220 rpm until logarithmic growth phase, after which they were infected with helper phage M13 to obtain a purified phage library of human Fab fragments.

Example 2: Production of RSV F protein and recombinant antibodies

2.1 Obtaining RSV F protein

Recombinant F protein of different viral subtypes, including the F protein of RSV subtype A strain A2 (F-A2, with the amino acid sequence defined in SEQ ID NO: 10) and the F protein of RSV subtype B strain 18537 (F-18537, with the amino acid sequence defined in SEQ ID NO: 11), were required for the production of monoclonal antibodies to the RSV F protein. Based on the two different RSV F protein subtypes, the inventors constructed mutant F proteins of the DS-Cav1 ¹⁵ pre-fusion structure, designated "RSV-DS-Cav1-A" (with the amino acid sequence defined in SEQ ID NO: 12) and "RSV-DS-Cav1-B" (with the amino acid sequence defined in SEQ ID NO: 13), respectively. Post-translational modifications (e.g., glycosylation, disulfide bonds, etc.) occur in the F protein. Therefore, the use of a mammalian cell expression system would be preferable to preserve the structure and function of the recombinant protein. Furthermore, the addition of a histidine tag (His tag, with the amino acid sequence defined in SEQ ID NO:14) to the C-terminus of the recombinant protein is preferable for purification of the recombinant protein and determination of the function of monoclonal antibodies.

The RSV-DS-Cav1-A and RSV-DS-Cav1-B genes were synthesized. The synthesized genes were cloned into a suitable eukaryotic expression vector ( e.g., pcDNA3.1 from Invitrogen Inc., etc.) using methods known in the field of molecular biology. HEK293 cells (e.g., HEK293F from Invitrogen Inc., etc.) were transfected with the resulting plasmids expressing the recombinant protein using liposomes (e.g., 293fectin from Invitrogen Inc., etc.) or another cationic transfection reagent (e.g., PEI, etc.). The cells were cultured in a suspended state in a serum-free medium for 3-4 days. The cell culture supernatant was then collected by centrifugation and other means. The recombinant protein in the supernatant was subjected to a one-step purification using a metal-chelate affinity chromatography column (e.g., HisTrap FF, manufactured by G-E Inc.). The storage solution of the recombinant protein was replaced with phosphate-buffered saline (PBS solution) (pH 7.0) or other suitable buffers using a desalting column (e.g., Hitrap desaulting, manufactured by G-E Inc.). If necessary, the sample can be sterilized by filtration and then stored in aliquots at -20°C.

2.2 Production of recombinant antibodies

Nucleotide sequences encoding the variable region of the heavy chain and variable region of the light chain of antibodies were cloned into eukaryotic expression vectors (e.g., pcDNA3.1 from Invitrogen Inc., etc.) and fused with nucleotide sequences encoding the constant region of the heavy chain and constant region of the light chain, respectively, using methods known in molecular biology. Complete antibodies were expressed in combination. The constant region of the heavy chain of the antibodies may be of the human IgG1 subtype (with the amino acid sequence defined in SEQ ID NO: 15), the IgG1-YTE mutant of the human IgG1 subtype (with the amino acid sequence defined in SEQ ID NO: 16), the mouse IgG2a subtype (with the amino acid sequence defined in SEQ ID NO: 17), while the constant region of the light chain of the antibodies may be of the human kappa subtype (with the amino acid sequence defined in SEQ ID NO: 18), the human lambda subtype (with the amino acid sequence defined in SEQ ID NO: 19), the mouse kappa subtype (with the amino acid sequence defined in SEQ ID NO: 20) or the mouse lambda subtype (with the amino acid sequence defined in SEQ ID NO: 21).

HEK293 cells (e.g., HEK293F from Invitrogen Inc.) were transfected with plasmids expressing the resulting recombinant antibody using liposomes (e.g., 293fectin from Invitrogen Inc., etc.) or other transfection reagents (e.g., PEI, etc.). The cells were cultured suspended in serum-free medium for 3-5 days. The cell culture supernatant was then collected by centrifugation and other means. The recombinant antibodies in the supernatant were subjected to a one-step purification using an A/G protein affinity chromatography column (e.g., Mabselect SURE from GE Inc., etc.). The storage solution for the recombinant protein was exchanged for PBS (pH 7.0) or other suitable buffers using a desalting column (e.g., Hitrap desaulting column from GE Inc.). If necessary, the sample can be sterilized by filtration and then stored in aliquots at -20°C.

Example 3: Screening a library of fully human recombinant Fab fragments with the RSV F protein

According to the reference document (for detailed experimental protocols, incorporated herein by reference in their entirety, see Chinese Patent Application No. 201510097117.0), the fully human recombinant Fab library constructed in Example 1 was screened by the solid phase method (for the experimental protocol, see “Phage Display: A Practical Approach, Clackson, T. (USA) and Lowman, H. B. (USA) (ed.)”, translated by Lan Ma et al., Chemical Industry Press, May 2008) using the recombinant RSV F protein obtained in Example 2. Four to six rounds of screening were carried out by Binding, elution, neutralization, infection, and amplification were performed. As a result, two Fab antibodies, R3B1h1 and R22B1, were obtained that specifically bind to the RSV F protein.

Example 4: Detection of recombinant monoclonal antibodies to the RSV F protein

The nucleotide sequences encoding the light and heavy chains of two R3B1h1 and R22B1 molecules were cloned into eukaryotic expression vectors using known molecular biology methods to produce monoclonal antibodies with a recombinant version of human IgG1-κ. In this case, the human monoclonal antibody to RSV palivizumab (Hu1129) (with the amino acid sequence of the heavy chain variable region defined in SEQ ID NO: 22 and the amino acid sequence of the light chain variable region defined in SEQ ID NO: 23) was obtained in accordance with patent No. US 7,704,505 B2, the human monoclonal antibody to RSV nirsevimab (MEDI8897) (with the amino acid sequence of the heavy chain variable region defined in SEQ ID NO: 24 and the amino acid sequence of the light chain variable region defined in SEQ ID NO: 25) in accordance with patent No. US 1,118,628 B2, and the human monoclonal antibody to RSV clesrovimab (RB1) (with the amino acid sequence of the heavy chain variable region defined in SEQ ID NO: 26, and the amino acid sequence of the light chain variable region defined in SEQ ID NO: 27) in accordance with US Patent No. 9963500 B2, wherein these three antibodies were used as positive control antibodies.

4.1 Confirmation of the binding activity of recombinant monoclonal antibodies to the RSV F protein

96-well ELISA plates were coated with the obtained RSV-DS-Cav1-A and RSV-DS-Cav1-Bat at a concentration of 3 μg/ml, 100 μl per well, and incubated at 4°C overnight, respectively. After blocking the plates with a blocking solution (milk-based solution of PBS-0.1% and Tween 20-3% (milk PBST-3%)) at 37°C for 1 hour, each recombinant monoclonal antibody to the RSV F protein was added to the corresponding plate and subjected to binding at 37°C for 1 hour. ELISA plates were washed with a buffer solution based on PBS-0.1% and Tween 20 (PBST solution), mouse antibody with horseradish peroxidase (HRP) to human IgG (Bioss, bsm-0297M-HRP) was added and subjected to binding at 37°C for 1 hour. ELISA plates were washed with a buffer solution of PBST. Color developing solution of OPD (ortho-phenylenediamine) substrate was added. After incubation for 5-10 minutes, color development was stopped with 1M H ₂ SO ₄ solution. Optical density values were measured at two wavelengths of 492 nm/630 nm using a microplate photometer. The results of the ELISA analysis (Fig. 3) indicated that the recombinant monoclonal antibodies R3B1h1 and R22B1 to the RSV F protein exhibit approximately equal activity in binding the two recombinant proteins RSV-DS-Cav1-A and RSV-DS-Cav1-B.

4.2 Epitope analysis of recombinant monoclonal antibodies to the RSV F protein

96-well ELISA plates were coated with recombinant RSV-DS-Cav1-B protein (1 μg/ml, 100 μl/well) and incubated overnight at 4°C. The plates were blocked with blocking solution (PBST-3% milk) at 37°C for 1 h. Gradient dilution of recombinant monoclonal antibodies to RSV F protein (R3B1h1/R22B1/klesrovimab) with each of the purified phages to RSV F protein (R3B1h1/R22B1/klesrovimab/nirsevimab) was performed at a fixed concentration (1× ¹⁰¹¹ CFU/ml), respectively. The initial concentration of antibodies was 200 μg/ml, diluted 3-fold, and the total number of concentration gradients was 10. The diluted antibodies were added to blocked 96-well ELISA plates at a concentration of 100 μl per well, and the plates were incubated at 37°C for 1 hour. The ELISA plates were washed with PBST buffer, followed by the addition of the secondary antibody with HRP to M13 (Sino Biological Inc., 11973-MM05T-H), and the plates were incubated at 37°C for 1 hour. The ELISA plates were washed with PBST solution, and the color developing solution of the OPD substrate was added. After incubation for 5-10 minutes, color development was stopped with a 1M solution of H ₂ SO ₄ . The optical density values were measured at two wavelengths of 492 nm/630 nm using a microplate reader. The ELISA results are shown in Fig. 4. Monoclonal antibody R3B1h1 could block the binding of R22B1/nirsevimab phages to the recombinant RSV-DS-Cav1-B protein, but did not affect the binding of klesrovimab phage to the recombinant RSV-DS-Cav1-B protein (Fig. 4A). Monoclonal antibody R22B1 could block the binding of R3B1h1/nirsevimab phages to the recombinant RSV-DS-Cav1-B protein, but did not affect the binding of klesrovimab phage to the recombinant RSV-DS-Cav1-B protein (Fig. 4B). Monoclonal antibody clesrovimab failed to block the binding of R3B1h1/R22B1/nirsevimab phages to recombinant RSV-DS-Cav1-B protein (Fig. 4C).

4.3 Affinity analysis of recombinant monoclonal antibodies to the RSV F protein

The affinity of monoclonal antibodies to the RSV F protein was determined by surface plasmon resonance using a Biacore T200 instrument. Relevant reagents and consumables, including the amine coupling kit (BR-1000-50), human antibody capture kit (BR-1008-39), CM5 chip (BR100012), and 10×HBS-EP buffer (pH 7.4, BR100669), were purchased from GE Healthcare. The carboxylated CM5 chip was surface activated with 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) according to the kit instructions. Anti-human IgG antibody (Fc fragment) (immobilized antibody) was diluted to a concentration of 25 μg/mL with 10 mM sodium acetate (pH 5.0) and then injected at a flow rate of 10 μL/min to achieve a coupling rate of approximately 10,000 response units (RU). Following injection of the immobilized antibody, 1 M ethanolamine was injected to block unreacted groups. For kinetic measurements, monoclonal antibodies to the RSV F protein were diluted to a concentration of 0.5-1 μg/mL and injected at a flow rate of 10 μL/min to ensure antibody capture of approximately 80 RU by the anti-human Fc fragment antibody. RSV-DS-Cav1-A or RSV-DS-Cav1-B were diluted with a series of concentration gradients (e.g., 6.17 nmol, 18.5 nmol, 55.6 nmol, 166.7 nmol, 500 nmol) and injected at 25°C from low to high concentrations at a flow rate of 30 μL/min, with an association time of 90 seconds and a dissociation time of 3600 seconds. 3 M _MgCl2 solution was injected at a flow rate of 10 μL/min for 30 seconds to regenerate the chip surface. The association rate ( _Kon ) and dissociation rate ( _Koff ) were calculated by fitting the sensor-derived association and dissociation patterns to a 1:1 association model using the Biacore T200 version 3.0 evaluation software. The equilibrium constant for dissociation (K _D ) was calculated as the ratio K _off /K _on . The comparison results are presented in Tables 1 and 2.

Table 1. Affinity constants for binding of recombinant monoclonal antibodies to RSV F protein to RSV-DS-Cav1-A

K _on (1/MS) K _off (1/S) K _D (M) Palivizumab 3.158E+4 8.197E-5 2.595E-9 Klesrovimab 2.001E+4 3.515E-4 1.756E-8 Nirsevimab 3.149E+4 1.502E-5 4.770E-10 R3B1h1 3.184E+4 2.370E-5 6.213E-10 R22B1 3.991E+4 1.411E-5 6.213E-10

Table 2. Affinity constants for binding of recombinant monoclonal antibodies to RSV F protein to RSV-DS-Cav1-B

K _on (1/MS) K _off (1/S) K _D (M) Palivizumab 1.360E+5 2.001E-4 1.471E-9 Klesrovimab 9.171E+6 8.208E-5 8.951E-12 Nirsevimab 1.082E+5 3.327E-4 3.075E-9 R3B1h1 2.286E+5 8.093E-5 3.541E-10 R22B1 3.577E+5 1.010E-4 2.824E-10

4.4 Inhibition of RSV infection of Hep-2 cells with recombinant monoclonal antibodies to the RSV F protein

4.4.1 Inhibition of RSV A2 strain infection of Hep-2 cells with recombinant monoclonal antibodies to RSV F protein

Monoclonal antibodies to RSV F protein (R3B1h1, R22B1, nirsevimab, clesrovimab, and palivizumab) and negative isotype control antibody were diluted in Hep-2 cell maintenance medium (Dulbecco's modified Eagle's medium (DMEM) + fetal bovine serum (FBS) 2% + penicillin/synthomycin (P/S) 1% + glutamate 2 mM) at an initial concentration of 66.7 nM, with a 5-fold gradient, with a total number of concentration gradients equal to 10. Diluted antibodies were added to 96-well sterile, fully permeable cell culture plates. One frozen solution of RSV type A2 was placed in a P2 laboratory microbiological safety cabinet and slowly thawed, after which it was diluted to a concentration of 1.5 × 10 ³ PFU/mL with Hep-2 cell maintenance medium. The diluted virus solution was added to the diluted antibody at 70 μL per well and thoroughly mixed. At the same time, separate wells were prepared for control cells (Hep-2 cell maintenance medium at 140 μL per well) and “cell + RSV strain A2” wells (Hep-2 cell maintenance medium at 70 μL per well + diluted RSV strain A2 at 70 μL per well). After adding all samples, the plates were covered, placed in a cell culture chamber (at 37 ± 1 °C, 5 ± 1% CO ₂ ) and neutralized for 1 hour. Logarithmic-phase Hep-2 cells were harvested, digested, centrifuged, and diluted. The counted cell suspension was then diluted with Hep-2 cell maintenance medium to obtain a single cell suspension at a concentration of 4.3× ¹⁰⁵ cells/mL. The cells were seeded in 96-well cell culture plates, which were neutralized at 70 μL per well and incubated in a cell culture incubator (37±1°C, 5±1% _CO2 ) for 60-72 h. After viral infiltration, the cells were washed twice with PBS and incubated for 30 min in a refrigerator at 2~8°C with 80% acetone. The 80% acetone was then removed. The cells were washed twice with PBS, then treated with the detection antibody palivizumab (100 μl/well) diluted to a concentration of 10 μg/ml with PBS, and incubated at 37°C in the dark for 60 minutes. After removing the primary antibody, the cells were washed twice with PBS. Next, FITC-labeled goat anti-human IgG (ZSGB-BIO, ZF-0308) diluted with PBS (1:400) were added at a concentration of 100 μl/well and incubated at 37°C in the dark for 40 minutes. After removing the secondary antibody, the cells were washed twice with PBS. Chemiluminescence values were determined by a rapid fluorescence inhibition test (RFFIT test) using a multifunctional microplate reader (SpectraMax I3). The results (Fig. 5) indicated that R3B1h1 had the best activity, being approximately 5 times more active than nirsevimab, approximately 12 times more active than clesrovimab, and approximately 410 times more active than palivizumab based on the comparison of IC ₅₀ values (Table 3).

Table 3. Half-inhibitory concentrations of monoclonal antibodies to the RSV F protein that inhibit infection of Hep-2 cells by the RSV strain A2

Antibody name IC ₅₀ (nmol) R3B1h1 0.005162 R22B1 0.07067 Nirsevimab 0.02306 Klesrovimab 0.06219 Palivizumab 2.12 Isotype control antibody /

4.4.2 Inhibition of RSV strain 18537 infection of Hep-2 cells with recombinant monoclonal antibodies to RSV F protein

The neutralizing activity of recombinant monoclonal antibodies to RSV F protein (R3B1h1, R22B1, nirsevimab, clesrovimab, and palivizumab) against RSV strain 18537 was determined according to the protocol described in Section 4.4.1. The results (Fig. 6) indicated that R3B1h1 had the best activity, which was approximately 18-fold higher than nirsevimab, approximately 12-fold higher than clesrovimab, and approximately 545-fold higher than palivizumab based on the comparison of IC ₅₀ values (Table 4).

Table 4. Half-inhibitory concentrations of monoclonal antibodies to the RSV F protein that inhibit infection of Hep-2 cells with RSV strain 18537

Antibody name IC ₅₀ (nmol) R3B1h1 0.007595 R22B1 0.1111 Nirsevimab 0.1392 Klesrovimab 0.08802 Palivizumab 4,146 Isotype control antibody /

4.4.3 Inhibition of Hep-2 cell infection with recombinant monoclonal antibodies to RSV F protein isolated from clinical RSV samples

The neutralizing activity of recombinant monoclonal antibodies to the RSV F protein (R3B1h1, R22B1, nirsevimab, clesrovimab, and palivizumab) against RSV isolated from clinical samples was determined according to the protocol described in Section 4.4.1.

A total of 11 strains isolated from clinical specimens collected during the 2021 RSV epidemic season were obtained from the Guangzhou Women's and Children's Health Center. Of these, nine strains belonged to subtype A (numbers 2033, 2064, 2066, 2406, 2410, 2416, 2425, 2430, and 2438, respectively), and two strains belonged to subtype B (numbers 2063 and 2427, respectively). The results (Fig. 7) indicate that both R3B1h1 and R22B1 are capable of successfully neutralizing clinical RSV strains.

Sequence information

SEQ ID NO:1

CGAATAAATACCTGTGACGGAAGATCACTTCGCAGAATAAATAAATCCTGGTGTCCCTGTTGATACCGGGAAGCCCTGGGCCAACTTTTGGCGAAAATGAGACGTTGATCGGCACGTAAGAGGTTCCAACTTTCACCATAATGAAATAAGATCACTACCGGGCGTATTTTTTGAGTTATCGAGATTTTCA

SEQ ID NO:2

AAATAATGATTTTATTTTGACTGATAGTGACCTGTTCGTTGCAACACATTGATGAGCAATGCTTTTTTATAATGCCAACTTTGTACAAAAAAGCTGAACGAGAAACGTAAAATGATATAAATTCAATATATTAAATTAGATTTTGCATAAAAAACAGACTACATAATACTGTAAAACACAACATATCCAGTCACTATGAATCAACTACTTAGATGGTATTAGTGACCTGTA

SEQ ID NO:3

GATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTTTTGCCCTGTAAACGAAAAAACCACCTGGGGAGGTGGTTTGATCGAAGGTTAAGTCAGTTGGGGAACTGCTTAACCGTGGTAACTGGTTTTTCGCAGAGCACAGCAACCAAATCTGTCCTTCCAGTGTAGCCGGACTTTGG CGCACACTTCAAGAGCAACCGCGTGTTTAGCTAAACAAATCCTCTGCGAACTCCCAGTTACCAATGGCTGCTGCCAGTGGCGTTTTACCGTGCTTTTCCGGGTTGGACTCAAGTGAACAGTTACCGGATAAGGCGCAGCAGTCGGGCTGAACGGGGAGTTCTTGCTTACAGCCCAGCTTGGAGCG AACGACCTACACCGAGCCGAGATACCAGTGTGTGAGCTATGAGAAAGCGCCACACTTCCCGTAAGGGAGAAAGGCGGAACAGGTATCCGGTAAACGGCAGGGTCGGAACAGGAGAGCGCAAGAGGGAGCGACCCGCCGGAAACGGTGGGGATCTTTAAGTCCTGTCGGGTTTCGCCCGTACTGTC AGATTCATGGTTGAGCCTCACGGCTCCCACAGATGCACCGGAAAAGCGTCTGTTTATGTGAACTCTGGCAGGAGGGCGGAGCCTATGGAAAAACGCCACCGGCGCGGCCCTGCTGTTTTGCCTCACATGTTAGTCCCCTGCTTATCCACGGAATCTGTGGGTAACTTTGTATGTGTCCGCAGCGC

SEQ ID NO:4

ACAAGTTTGTACAAAAAAGCAGGCT

SEQ ID NO: 5

ATGGAGAAAAAAATCACTGGATATACCACCGTTGATATATCCCAATGGCATCGTAAAGAACATTTTTGAGGCATTTCAGTCAGTTGCTCAATGTACCTATAACCAGACCGTTCAGCTGGATATTACGGCCTTTTTAAAGACCGTAAAGAAAAATAAGCACAAGTTT TATCCGGCCTTTATTCACATTCTTGCCCGCCTGATGAATGCTCATCCGGAATTCCGTATGGCAATGAAAGACGGTGAGCTGGTGATATGGGATAGTGTTCACCCTTGTTACACCGTTTTTCCATGAGCAAACTGAAACGTTTTCATCGCTCTGGAGTGAATACCAC GACGATTTCCGGCAGTTTCTACACATATATTCGCAAGATGTGGCGTGTTACGGTGAAAACCTGGCCTATTTCCCTAAAGGGTTTATTGAGAATATGTTTTTCGTCTCAGCCAATCCCTGGGTGAGTTTCACCAGTTTTGATTTAAACGTGGCCAATATGGACAAC TTCTTCGCCCCCGTTTTCACTATGGGCAAATATTATACGCAAGGCGACAAGGTGCTGATGCCGCTGGCGATTCAGGTTCATCATGCCGTCTTTGATGGCTTCCATGTCGGCAGAATGCTTAATGAATTACAACAGTACTGCGATGAGTGGCAGGGCGGGGCGTAA

SEQ ID NO:6

TCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAAC ATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAA GATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTT GAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTT GCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACT CTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACT GGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATAT ATACTTTAGATTGATTTAAAACTTCATTTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTC

SEQ ID NO:7

CTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTAAGTTAGCTCACTCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATAATGTGTGGAATTGTGAGCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGGATTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCC TGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGCGCTTTGCCTGGTTTCCGGCACCAGAAGCGGTGCCGGAAAGCTGGCTGGAGTGCGATCTTCCTGAGGCCGATACTGTCGTCGTCCCCTCAAACTGGCAGATG CACGGTTACGATGCGCCCATCTACACCAACGTGACCTATCCCATTACGGTCAATCCGCCGTTTGTTCCCACGGAATCCGACGGGTTGTTACTCGCTCACATTTAATGTTGATGAAAGCTGGCTACAGGAAGGCCAGACGCGAATTATTTTTGATGGCGTCGGGATCTGATCCGGATTTACTAACTGGAAGAGGCACTAAATGGGAAGAAGGCGAAGTC ATGAGCGCCGGGATTTACCCCCTAACCTTTATATAAGAAACAATGGATATTACTGCTACAGGGACCCAAGGACGGGTAAAGAGTTTGGATTAGGCAGAGACAGGCGAATCGCAATCACTGAAGCTATACAGGCCAACATTGAGTTATTTTCAGGACACAAACACAAGCCTCTGACAGCGAGAATCAACAGTGATAATTCCGTTACGTTACATTCATGGCT TGATCGCTACGAAAAAATCCTGGCCAGCAGAGGAATCAAGCAGAAGACACTCATAAATTACATGAGCAAAATTAAAGCAATAAGGAGGGGTCTGCCTGATGCTCCACTTGAAGACATCACCACAAAAGAAATTGCGGCAATGCTCAATGGATACATAGACGAGGGCAAGGCGGCGTCAGCCAAGTTAATCAGATCAACACTGAGCGATGCATTCCGAGAG GCAATAGCTGAAGGCCATATAACAACAAACCATGTCGCTGCCACTCGCGCAGCAAAATCAGAGGTAAGGAGATCAAGACTTACGGCTGACGAATACCTGAAAATTTATCAAGCAGCAGAATCATCACCATGTTGGCTCAGACTTGCAATGGAACTGGCTGTTGTTACCGGGCAACGAGTTGGTGATTTATGCGAAATGAAGTGGTCTGATATCGTAGATG GATATCTTTATGTCGAGCAAAGCAAAACAGGCGTAAAAATTGCCATCCCAACAGCATTGCATATTGATGCTCTCGGAATATCAATGAAGGAAACACTTGATAAATGCAAAGAGATTCTTGGCGGAGAAACCATAATTGCATCTACTCGTCGCGAACCGCTTTCATCCGGCACAGTATCAAGGTATTTTATGCGCGCACGAAAAGCATCAGGTCTTTCCTT CGAAGGGGATCCGCCTACCTTTCACGAGTTGCGCAGTTTGTCTGCAAGACTCTATGAGAAGCAGATAAGCGATAAGTTTGCTCAACATCTTCTCGGGCATAAGTCGGACACCATGGCATCACAGTATCGTGATGACAGAGGCAGGGAGTGGGACAAAATTGAAATCAAATAACTAGCATAACCCCTTGGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTG

SEQ ID NO:8

AAATAATGATTTTATTTTGACTGATAGTGACCTGTTCGTTGCAACACATTGATGAGCAATGCTTTTTTATAATGCCAACTTTGTACAAAAAAGCAGGCT

SEQ ID NO:9

ACAAGTTTGTACAAAAAAGCTGAACGAGAAACGTAAAATGATATAAATATCAATATATTAAATTAGATTTTGCATAAAAAACAGACTACATAATACTGTAAAACACAACATATCCAGTCACTATGAATCAACTACTTAGATGGTATTAGTGACCTGTA

SEQ ID NO:10

MELLILKANAITTILTAVTFCFASGQNITEEFYQSTCSAVSKGYLSALRTGWYTSVITIELSNIKENKCNGTDAKVKLIKQELDKYKNAVTELQLLMQSTPPTNNRARRELPRFMNYTLNNAKKTNVTLSKKRKRRFLGFLLG VGSAIASGVAVSKVLHLEGEVNKIKSALLSTNKAVSLSNGVSVLTSKVLDLKNYIDKQLLPIVNKQSCSISNIETVIEFQQKNNRLLEITREFSVNAGVTTTPVSTYMLTNSELLSLINDMPITNDQKKLMSNNVQIVRQQSYS IMSIIKEEVLAYVVQLPLYGVIDTPCWKLHTSPLCTTNTKEGSNICLTRTDRGWYCDNAGSVSFFPQAETCKVQSNRVFCDTMNSLTLPSEINLCNVDIFNPKYDCKIMTSKTDVSSSVITSLGAIVSCYGKTKCTASNKNRG IIKTFSNGCDYVSNKGMDTVSVGNTLYYVNKQEGKSLYVKGEPIINFYDPLVFPSDEFDASISQVNEKINQSLAFIRKSDELLHNVNAGKSTTNIMITTIIIVIIVILLSLIAVGLLLYCKARSTPVTLSKDQLSGINNIAFSN

SEQ ID NO:11

MELLIHRSSAIFLTLAVNALYLTSSQNITEEFYQSTCSAVSRGYFSALRTGWYTSVITIELSNIKETKCNGTDTKVKLIKQELDKYKNAVTELQLLMQNTPAANNRARREAPQYMNYTINTTKNLNVSISKKRKRRFLGFLLG VGSAIASGIAVSKVLHLEGEVNKIKNALLSTNKAVVSLSNGVSVLTSKVLDLKNYINNRLLPIVNQQSCRISNIETVIEFQQMNSRLLEITREFSVNAGVTTPLSTYMLTNSELLSLINDMPITNDQKKLMSSNVQIVRQQSYS IMSIIKEEVLAYVVQLPIYGVIDTPCWKLHTSPLCTTNIKEGSNICLTRTDRGWYCDNAGSVSFFPQADTCKVQSNRVFCDTMNSLTLPSEVSLCNTDIFNSKYDCKIMTSKTDISSSVITSLGAIVSCYGKTKCTASNKNRG IIKTFSNGCDYVSNKGVDTVSVGNTLYYVNKLEGKNLYVKGEPIINYYDPLVFPSDEFDASISQVNEKINQSLAFIRRSDELLHNVNTGKSTTNIMITTIIIVIIVVLLSLIAIGLLLYCKAKNTPVTLSKDQLSGINNIAFSK

SEQ ID NO:12

MELLILKANAITTILTAVTFCFASGQNITEEFYQSTCSAVSKGYLSALRTGWYTSVITIELSNIKENKCNGTDAKVKLIKQELDKYKNAVTELQLLMQSTPPTNNRARRELPRFMNYTLNNAKKTNVTLSKKRKRR FLGFLLGVGSAIASGVAVCKVLHLEGEVNKIKSALLSTNKAVVSLSNGVSVLTFKVLDLKNYIDKQLLPILNKQSCSISNIETVIEFQQKNNRLLEITREFSVNAGVTTTPVSTYMLTNSELLSLINDMPITNDQKK LMSNNVQIVRQQSYSIMCIIKEEVLAYVVQLPLYGVIDTPCWKLHTSPLCTTNTKEGSNICLTRTDRGWYCDNAGSVSFFPQAETCKVQSNRVFCDTMNSLTLPSEINLCNVDIFNPKYDCKIMTSKTDVSSSVIT SLGAIVSCYGKTKCTASNKNRGIIKTFSNGCDYVSNKGMDTVSVGNTLYYVNKQEGKSLYVKGEPIINFYDPLVFPSDEFDASISQVNEKINQSLAFIRKSDELLSAIGGYIPEAPRDGQAYVRKDGEWVLLSTFL

SEQ ID NO:13

MELLIHRSSAIFLTLAVNALYLTSSQNITEEFYQSTCSAVSRGYFSALRTGWYTSVITIELSNIKETKCNGTDTKVKLIKQELDKYKNAVTELQLLMQNTPAANNRARREAPQYMNYTINTTKNLNVSISKKRKRR FLGFLLGVGSAIASGIAVCKVLHLEGEVNKIKNALLSTNKAVVSLSNGVSVLTFKVLDLKNYINNRLLPILNQQSCRISNIETVIEFQQMNSRLLEITREFSVNAGVTTPLSTYMLTNSELLSLINDMPITNDQKK LMSSNVQIVRQQSYSIMCIIKEEVLAYVVQLPIYGVIDTPCWKLHTSPLCTTNIKEGSNICLTRTDRGWYCDNAGSVSFFPQADTCKVQSNRVFCDTMNSLTLPSEVSLCNTDIFNSKYDCKIMTSKTDISSSVIT SLGAIVSCYGKTKCTASNKNRGIIKTFSNGCDYVSNKGVDTVSVGNTLYYVNKLEGKNLYVKGEPIINYYDPLVFPSDEFDASISQVNEKINQSLAFIRRSDELLSAIGGYIPEAPRDGQAYVRKDGEWVLLSTFL

SEQ ID NO:14

HHHHHH

SEQ ID NO:15

ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

SEQ ID NO:16

ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

SEQ ID NO:17

AKTTAPSVYPLAPVCGDTTGSSVTLGCLVKGYFPEPVTLTWNSGSLSSGVHTFPAVLQSDLYTLSSSVTVTSSTWPSQSITCNVAHPASSTKVDKKIEPRGPTIKPCPPCKCPAPNLLGGPSVFIFPPKIKDVLMISLSPIVTCVVVDVSEDDPDVQISWFVNNV EVHTAQTQTHREDYNSTLRVVSALPIQHQDWMSGKEFKCKVNNKDLPAPIERTISKPKGSVRAPQVYVLPPPEEEMTKKQVTLTCMVTDFMPEDIYVEWTNNGKTELNYKNTEPVLDSDGSYFMYSKLRVEKKNWVERNSYSCSVVHEGLHNHHTTKSFSRTPGK

SEQ ID NO:18

RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

SEQ ID NO:19

GQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS

SEQ ID NO:20

RADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSSPIVKSFNRNEC

SEQ ID NO:21

GQPKSSPSVTLFPPSSEELETNKATLVCTITDFYPGVVTVDWKVDGTPVTQGMETTQPSKQSNNKYMASSYLTLTARAWERHSSYSCQVTHEGHTVEKSLSRADCS

SEQ ID NO:22

QVTLRESGPALVKPTQTLTLTCTFSGFSLSTSGMSVGWIRQPPGKALEWLADIWWDDKKDYNPSLKSRLTISKDTSKNQVVLKVT NMDPADTATYYCARSMITNWYFDVWGAGTTVTVSS

SEQ ID NO:23

DIQMTQSPSTLSASVGDRVTITCKCQLSVGYMHWYQQKPGKAPKLWIYDTSKLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCFQGSGYPFTFGGGTKLEIK

SEQ ID NO:24

QVQLVQSGAEVKKPGSSVMVSCQASGGLLEDYIINWVRQAPGQGPEWMGGIIPVLGTVHYGPKFQGRVTITADESTDTAYMELSSLRSEDTAMYYCATETALVVSETYLPHYFDNWGQGTLVTVSS

SEQ ID NO:25

DIQMTQSPSSLSAAVGDRVTITCQASQDIVNYLNWYQQKPGKAPKLLIYVASNLETGVPSRFSGSGSGTDFSLTISSLQPEDVATYYCQQYDNLPLTFGGGTKVEIK

SEQ ID NO:26

EVQLVESGGGLVRPGRSLRLSCTVSGFSFDDSAMSWVRQAPGKGLEWISFIKSKTYGGTKEYAASVKGRFTISRDDSKNIAYLQMNSLKTEDTAVYYCTRGAPYGGNSDYYYGLDVWGQGTTVTVSS

SEQ ID NO:27

DIQMTQSPSSLSASVGDRVTITCRTSQDVRGALAWYQQKPGKAPKLLIFDASSLETGVPSRFSGSGSGTVFTLTISSLQPEDFAAYYCQQFLDFPFTFGQGTRLEIK

SEQ ID NO:28

QVQLVQSGAEVKKPGSSVKVSCKASGGLLEDYIINWVRQAPGQGLEWMGGIIPVLGTVHYGPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCATETAIVVTESYRPHFFDNWGQGTLVTVSS

SEQ ID NO:29

DIQMTQSPSSLSAsVGDRVTITCQASQDIVNYLNWYQQKPGKAPKLLIYVASNLETGVPSRFSGSGSGTDFtfTISSLQPEDiATYYCQEFAGLALNFGGGTKVEIK

SEQ ID NO:30

QVQLVQSGAEVKKPGSSVMVSCQASGDPLQNYIINWVRQAPGQGPEWMGAIIPALGSAHYGPKFQGRVTITADESTDTAYMELSSLRSEDTAMYYCATETAVIISESYLPHFFDNWGQGTLVTVSS

SEQ ID NO:31

DIQMTQSPSSLSAAVGDRVTITCQASQDIVNYLNWYQQKPGKAPKLLIYVASNLETGVPSRFSGSGSGTDFSLTISSLQPEDVATYYCQEYDSLALAFGGGTKVEIK

SEQ ID NO:32

DYIIN

SEQ ID NO:33

GIIPVLGTVHYGPKFQG

SEQ ID NO:34

ETAIVVTESYRPHFFDN

SEQ ID NO:35

QASQDIVNYLN

SEQ ID NO:36

VASNLET

SEQ ID NO:37

QEFAGLALN

SEQ ID NO:38

NYIIN

SEQ ID NO:39

AIIPALGSAHYGPKFQG

SEQ ID NO:40

ETAVIISESYLPHFFDN

SEQ ID NO:43

QEYDSLALA

SEQ ID NO:44

TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGT CCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACC GGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCC GGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA

SEQ ID NO:45

CGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAACTTGATTAGGGTGATGGTT CACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAAATTTAACGCGAATTTTAACAAAATATTAACGCTTACAATTT

SEQ ID NO:46

GGTGGCGGCTCCGGTTCCGGTGATTTTGATTATGAAAAAATGGCAAACGCTAATAAGGGGGCTATGACCGAAAATGCCGATGAAAACGCGCTACAGTCTGACGCTAAAGGCAAACTTGA TTCTGTCGCTACTGATTACGGTGCTGCTATCGATGGTTTCATTGGTGACGTTTCCGGCCTTGCTAATGGTAATGGTGCTACTGGTGATTTTGCTGGCTCTAATTCCCAAATGGCTCAAG TCGGTGACGGTGATAATTCACCTTTAATGAATAATTTCCGTCAATATTTACCTTCCCTCCCTCAATCGGTTGAATGTCGCCCTTTTGTCTTTGGCGCTGGTAAACCATATGAATTTTCT ATTGATTGTGACAAAATAAACTTATTCCGTGGTGTCTTTGCGTTTCTTTTATATGTTGCCACCTTTATGTATGTATTTTCTACGTTTGCTAACATACTGCGTAATAAGGAGTCTTGATAA

Sources:

1. JAMA Pediatr. 2016 Mar;170(3):267-87.

2. Lancet Glob Health. 2017 Oct;5(10): e984-e991.

3. J Gen Virol. 2017 Dec;98(12):2912-2913.

4. J Gen Virol. 1985 Oct;66(Pt 10):2111-24.

5. J Virol. 2008 Mar;82(5):2040-55.

6. J Virol. 1987 Oct;61(10):3163-6.

7. Proc Natl Acad Sci U S A. 1984 Dec;81(24):7683-7.

8. Virus Res. 2000 Jun;68(1):25-33.

9. Virology. 1993 Nov;197(1):1-11.

10. Gene. 1994 Dec 30;151(1-2):109-13.

11. Kabat, “Sequences of Proteins of Immunological Interest,” National Institutes of Health, Bethesda, Md. (1991).

12. A1-Lazikani, et al., J. Mol. Biol. 273:927-948 (1997).

13. Martin, et al., Proc. Natl. Acad. Sci.USA86:9268-9272 (1989).

14. Annu Rev Biochem. 1989; 58:913-49.

15. Science. 2013 Nov 1; 342(6158):592-8.

--->

<?xml version="1.0" encoding="UTF-8"?>

<!DOCTYPE ST26SequenceListing PUBLIC "-//WIPO//DTD Sequence Listing

1.3//EN" "ST26SequenceListing_V1_3.dtd">

<ST26SequenceListing originalFreeTextLanguageCode="en"

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<ApplicantFileReference>23C83147RU</ApplicantFileReference>

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<IPOfficeCode>CN</IPOfficeCode>

<ApplicationNumberText>202210871810.9</ApplicationNumberText>

<FilingDate>2022-07-22</FilingDate>

</EarliestPriorityApplicationIdentification>

<ApplicantName languageCode="en">Beijing Wisdomab Biotechnology Co.,

Ltd.</ApplicantName>

<InventionTitle languageCode="en">ANTI-RESPIRATORY SYNCYTIAL VIRUS

NEUTRALIZING ANTIBODY AND USE THEREOF</InventionTitle>

<SequenceTotalQuantity>46</SequenceTotalQuantity>

<SequenceData sequenceIDNumber="1">

<INSDSeq>

<INSDSeq_length>190</INSDSeq_length>

<INSDSeq_moltype>DNA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

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<INSDFeature_key>source</INSDFeature_key>

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<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>genomic DNA</INSDQualifier_value>

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<INSDQualifier id="q47">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>Escherichia coli</INSDQualifier_value>

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</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>cgaataaatacctgtgacggaagatcacttcgcagaataaataaatcct

ggtgtccctgttgataccgggaagccctgggccaacttttggcgaaaatgagacgttgatcggcacgtaa

gaggttccaactttcaccataatgaaataagatcactaccgggcgtattttttgagttatcgagattttc

a</INSDSeq_sequence>

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<SequenceData sequenceIDNumber="2">

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<INSDQualifier_value>Lambda phage targeting

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aatatcaatatattaaattagattttgcataaaaaacagactacataatactgtaaaacacaacatatcc

agtcactatgaatcaactacttagatggtattagtgacctgta</INSDSeq_sequence>

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</SequenceData>

<SequenceData sequenceIDNumber="3">

<INSDSeq>

<INSDSeq_length>739</INSDSeq_length>

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cgtggtaactggttttcgcagagcacagcaaccaaatctgtccttccagtgtagccggactttggcgcac

acttcaagagcaaccgcgtgtttagctaaacaaatcctctgcgaactcccagttaccaatggctgctgcc

agtggcgttttaccgtgcttttccgggttggactcaagtgaacagttaccggataaggcgcagcagtcgg

gctgaacggggagttcttgcttacagcccagcttggagcgaacgacctacaccgagccgagataccagtg

tgtgagctatgagaaagcgccacacttcccgtaagggagaaaggcggaacaggtatccggtaaacggcag

ggtcggaacaggagcgcaagagggagcgacccgccggaaacggtggggatctttaagtcctgtcgggt

ttcgcccgtactgtcagattcatggttgagcctcacggctcccacagatgcaccggaaaagcgtctgttt

atgtgaactctggcaggagggcggagcctatggaaaaacgccaccggcgcggccctgctgttttgcctca

catgttagtcccctgcttatccacggaatctgtgggtaactttgtatgtgtccgcagcgc</INSDSeq_

sequence>

</INSDSeq>

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<SequenceData sequenceIDNumber="4">

<INSDSeq>

<INSDSeq_length>25</INSDSeq_length>

<INSDSeq_moltype>DNA</INSDSeq_moltype>

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</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>acaagtttgtacaaaaaagcaggct</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="5">

<INSDSeq>

<INSDSeq_length>660</INSDSeq_length>

<INSDSeq_moltype>DNA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

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<INSDFeature_location>1..660</INSDFeature_location>

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<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>genomic DNA</INSDQualifier_value>

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<INSDQualifier id="q51">

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<INSDQualifier_value>Escherichia coli</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>atggagaaaaaaatcactggatataccaccgttgatatatcccaatggc

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tattacggcctttttaaagaccgtaaagaaaaataagcacaagttttatccggcctttattcacattctt

gcccgcctgatgaatgctcatccggaattccgtatggcaatgaaagacggtgagctggtgatatgggata

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cgacgatttccggcagtttctacacatatattcgcaagatgtggcgtgttacggtgaaaacctggcctat

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atttaaacgtggccaatatggacaacttcttcgcccccgttttcactatgggcaaatattatacgcaagg

cgacaaggtgctgatgccgctggcgattcaggttcatcatgccgtctttgatggcttccatgtcggcaga

atgcttaatgaattacaacagtactgcgatgagtggcagggcggggcgtaa</INSDSeq_sequence>

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<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>genomic DNA</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q52">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>Escherichia coli</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>tcaggtggcacttttcggggaaatgtgcgcggaacccctatttgtttat

ttttctaaatacattcaaatatgtatccgctcatgagacaataaccctgataaatgcttcaataatattg

aaaaaggaagagtatgagtattcaacatttccgtgtcgcccttattcccttttttgcggcattttgcctt

cctgtttttgctcacccagaaacgctggtgaaagtaaaagatgctgaagatcagttgggtgcacgagtgg

gttacatcgaactggatctcaacagcggtaagatccttgagagttttcgccccgaagaacgttttccaat

gatgagcacttttaaagttctgctatgtggcgcggtattatcccgtattgacgccgggcaagagcaactc

ggtcgccgcatacactattctcagaatgacttggttgagtactcaccagtcacagaaaagcatcttacgg

atggcatgacagtaagagaattatgcagtgctgccataaccatgagtgataacactgcggccaacttact

tctgacaacgatcggaggaccgaaggagctaaccgcttttttgcacaacatgggggatcatgtaactcgc

cttgatcgttgggaaccggagctgaatgaagccataccaaacgacgagcgtgacaccacgatgcctgtag

caatggcaacaacgttgcgcaaactattaactggcgaactacttactctagcttcccggcaacaattaat

agactggatggaggcggataaagttgcaggaccacttctgcgctcggcccttccggctggctggtttatt

gctgataaatctggagccggtgagcgtgggtctcgcggtatcattgcagcactggggccagatggtaagc

cctcccgtatcgtagttatctacacgacggggagtcaggcaactatggatgaacgaaatagacagatcgc

tgagataggtgcctcactgattaagcattggtaactgtcagaccaagtttactcatatatactttagatt

gatttaaaacttcatttttaatttaaaaggatctaggtgaagatcctttttgataatctcatgaccaaaa

tcccttaacgtgagttttcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttc</INSD

Seq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="7">

<INSDSeq>

<INSDSeq_length>1760</INSDSeq_length>

<INSDSeq_moltype>DNA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..1760</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>genomic DNA</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q53">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>Lambda phage targeting

Escherichia</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>ctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgca

attaatgtaagttagctcactcattaggcaccccaggctttacactttatgcttccggctcgtataatgt

gtggaattgtgagcggataacaatttcacacaggaaacagctatgaccatgattacggattcactggccg

tcgttttacaacgtcgtgactgggaaaaccctggcgttacccaacttaatcgccttgcagcacatccccc

tttcgccagctggcgtaatagcgaagaggcccgcaccgatcgcccttcccaacagttgcgcagcctgaat

ggcgaatggcgctttgcctggtttccggcaccagaagcggtgccggaaagctggctggagtgcgatcttc

ctgaggccgatactgtcgtcgtcccctcaaactggcagatgcacggttacgatgcgcccatctacaccaa

cgtgacctatcccattacggtcaatccgccgtttgttcccacggagaatccgacgggttgttactcgctc

acatttaatgttgatgaaagctggctacaggaaggccagacgcgaattatttttgatggcgtcgggatct

gatccggatttactaactggaagaggcactaaatgggaagaaggcgaagtcatgagcgccgggatttacc

ccctaaccttttatataagaaacaatggatattactgctacagggacccaaggacgggtaaagagtttgga

ttaggcagagacaggcgaatcgcaatcactgaagctatacaggccaacattgagttattttcaggacaca

aacacaagcctctgacagcgagaatcaacagtgataattccgttacgttacattcatggcttgatcgcta

cgaaaaaatcctggccagcagaggaatcaagcagaagacactcataaattacatgagcaaaattaaagca

ataaggaggggtctgcctgatgctccacttgaagacatcaccacaaaagaaattgcggcaatgctcaatg

gatacatagacgaggcaaggcggcgtcagccaagttaatcagatcaacactgagcgatgcattccgaga

ggcaatagctgaaggccatataacaacaaaccatgtcgctgccactcgcgcagcaaaatcagaggtaagg

agatcaagacttacggctgacgaatacctgaaaatttatcaagcagcagaatcatcaccatgttggctca

gacttgcaatggaactggctgttgttaccgggcaacgagttggtgatttatgcgaaatgaagtggtctga

tatcgtagatggatatctttatgtcgagcaaagcaaaacaggcgtaaaaattgccatcccaacagcattg

catattgatgctctcggaatatcaatgaaggaaacacttgataaatgcaaagagattcttggcggagaaa

ccataattgcatctactcgtcgcgaaccgctttcatccggcacagtatcaaggtattttatgcgcgcacg

aaaagcatcaggtctttccttcgaaggggatccgcctacctttcacgagttgcgcagtttgtctgcaaga

ctctatgagaagcagataagcgataagtttgctcaacatcttctcgggcataagtcggacaccatggcat

cacagtatcgtgatgacagaggcagggagtgggacaaaattgaaatcaaataactagcataaccccttgg

ggcctctaaacgggtcttgaggggttttttg</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="8">

<INSDSeq>

<INSDSeq_length>99</INSDSeq_length>

<INSDSeq_moltype>DNA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..99</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>other DNA</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q54">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>aaataatgattttattttgactgatagtgacctgttcgttgcaacacat

tgatgagcaatgcttttttataatgccaactttgtacaaaaaagcaggct</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="9">

<INSDSeq>

<INSDSeq_length>158</INSDSeq_length>

<INSDSeq_moltype>DNA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..158</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>other DNA</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q55">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>acaagtttgtacaaaaaagctgaacgagaaacgtaaaatgatataaata

tcaatatattaaattagattttgcataaaaaacagactacataatactgtaaaacacaacatatccagtc

actatgaatcaactacttagatggtattagtgacctgta</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="10">

<INSDSeq>

<INSDSeq_length>574</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..574</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q56">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>strain A2 of subtype A

RSV</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>MELLILKANAITTILTAVTFCFASGQNITEEFYQSTCSAVSKGYLSALR

TGWYTSVITIELSNIKENKCNGTDAKVKLIKQELDKYKNAVTELQLLMQSTPPPTNNRARRELPRFMNYTL

NNAKKTNVTLSKKRKRRFLGFLLGVGSAIASGVAVSKVLHLEGEVNKIKSALLSTNKAVVSLSNGVSVLT

SKVLDLKNYIDKQLLPIVNKQSCSISNIETVIEFQQKNNRLLEITREFSVNAGVTTTPVSTYMLTNSELLS

LINDMPITNDQKKLMSNNVQIVRQQSYSIMSIIKEEVLAYVVQLPLYGVIDTPCWKLHTSPLCTTNTKEG

SNICLTRTDRGWYCDNAGSVSFFPQAETCKVQSNRVFCDTMNSLTLPSEINLCNVDIFNPKYDCKIMTSK

TDVSSSVITSLGAIVSCYGKTKCTASNKNRGIIKTFSNGCDYVSNKGMDTVSVGNTLYYVNKQEGKSLYV

KGEPIINFYDPLVFPSDEFDASISQVNEKINQSLAFIRKSDELLHNVNAGKSTTNIMITTIIIVIIVILL

SLIAVGLLLYCKARSTPVTLSKDQLSGINNIAFSN</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="11">

<INSDSeq>

<INSDSeq_length>574</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..574</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q57">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>strain 18537 of subtype B

RSV</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>MELLIHRSSAIFLTLAVNALYLTSSQNITEEFYQSTCSAVSRGYFSALR

TGWYTSVITIELSNIKETKCNGTDTKVKLIKQELDKYKNAVTELQLLMQNTPAANNRARREAPQYMNYTI

NTTKNLNVSISKKRKRRFLGFLLGVGSAIASGIAVSKVLHLEGEVNKIKNALLSTNKAVVSLSNGVSVLT

SKVLDLKNYINNRLLPIVNQQSCRISNIETVIEFQQMNSRLLEITREFSVNAGVTTPLSTYMLTNSELLS

LINDMPITNDQKKLMSSNVQIVRQQSYSIMSIIKEEVLAYVVQLPIYGVIDTPCWKLHTSPLCTTNIKEG

SNICLTRTDRGWYCDNAGSVSFFPQADTCKVQSNRVFCDTMNSLTLPSEVSLCNTDIFNSKYDCKIMTSK

TDISSSVITSLGAIVSCYGKTKCTASNKNRGIIKTFSNGCDYVSNKGVDTVSVGNTLYYVNKLEGKNLYV

KGEPIINYYDPLVFPSDEFDASISQVNEKINQSLAFIRRSDELLHNVNTGKSTTNIMITTIIIVIIVVLL

SLIAIGLLLYCKAKNTPVTLSKDQLSGINNIAFSK</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="12">

<INSDSeq>

<INSDSeq_length>544</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..544</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q58">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>MELLILKANAITTILTAVTFCFASGQNITEEFYQSTCSAVSKGYLSALR

TGWYTSVITIELSNIKENKCNGTDAKVKLIKQELDKYKNAVTELQLLMQSTPPPTNNRARRELPRFMNYTL

NNAKKTNVTLSKKRKRRFLGFLLGVGSAIASGVAVCKVLHLEGEVNKIKSALLSTNKAVVSLSNGVSVLT

FKVLDLKNYIDKQLLPILNKQSCSISNIETVIEFQQKNNRLLEITREFSVNAGVTTTPVSTYMLTNSELLS

LINDMPITNDQKKLMSNNVQIVRQQSYSIMCIIKEEVLAYVVQLPLYGVIDTPCWKLHTSPLCTTNTKEG

SNICLTRTDRGWYCDNAGSVSFFPQAETCKVQSNRVFCDTMNSLTLPSEINLCNVDIFNPKYDCKIMTSK

TDVSSSVITSLGAIVSCYGKTKCTASNKNRGIIKTFSNGCDYVSNKGMDTVSVGNTLYYVNKQEGKSLYV

KGEPIINFYDPLVFPSDEFDASISQVNEKINQSLAFIRKSDELLSAIGGYIPEAPRDGQAYVRKDGEWVL

LSTFL</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="13">

<INSDSeq>

<INSDSeq_length>544</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..544</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q59">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>MELLIHRSSAIFLTLAVNALYLTSSQNITEEFYQSTCSAVSRGYFSALR

TGWYTSVITIELSNIKETKCNGTDTKVKLIKQELDKYKNAVTELQLLMQNTPAANNRARREAPQYMNYTI

NTTKNLNVSISKKRKRRFLGFLLGVGSAIASGIAVCKVLHLEGEVNKIKNALLSTNKAVVSLSNGVSVLT

FKVLDLKNYINNRLLPILNQQSCRISNIETVIEFQQMNSRLLEITREFSVNAGVTTPLSTYMLTNSELLS

LINDMPITNDQKKLMSSNVQIVRQQSYSIMCIIKEEVLAYVVQLPIYGVIDTPCWKLHTSPLCTTNIKEG

SNICLTRTDRGWYCDNAGSVSFFPQADTCKVQSNRVFCDTMNSLTLPSEVSLCNTDIFNSKYDCKIMTSK

TDISSSVITSLGAIVSCYGKTKCTASNKNRGIIKTFSNGCDYVSNKGVDTVSVGNTLYYVNKLEGKNLYV

KGEPIINYYDPLVFPSDEFDASISQVNEKINQSLAFIRRSDELLSAIGGYIPEAPRDGQAYVRKDGEWVL

LSTFL</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="14">

<INSDSeq>

<INSDSeq_length>6</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..6</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q60">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>HHHHHH</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="15">

<INSDSeq>

<INSDSeq_length>330</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..330</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q61">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>Homo sapiens</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSG

VHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLG

GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVL

TVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD

IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

K</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="16">

<INSDSeq>

<INSDSeq_length>330</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..330</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q62">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>Homo sapiens</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSG

VHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLG

GPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVL

TVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD

IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

K</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="17">

<INSDSeq>

<INSDSeq_length>330</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..330</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q63">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>Mus musculus</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>AKTTAPSVYPLAPVCGDTTGSSVTLGCLVKGYFPEPVTLTWNSGSLSSG

VHTFPAVLQSDLYTLSSSVTVTSSTWPSQSITCNVAHPASSTKVDKKIEPRGPTIKPCPPCKCPAPNLLG

GPSVFIFPPKIKDVLMISLSPIVTCVVVDVSEDDPDVQISWFVNNVEVHTAQTQTHREDYNSTLRVVSAL

PIQHQDWMSGKEFKCKVNNKDLPAPIERTISKPKGSVRAPQVYVLPPPEEEMTKKQVTLTCMVTDFMPED

IYVEWTNNGKTELNYKNTEPVLDSDGSYFMYSKLRVEKKNWVERNSYSCSVVHEGLHNHHTTKSFSRTPG

K</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="18">

<INSDSeq>

<INSDSeq_length>107</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..107</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q64">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>Homo sapiens</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQS

GNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC</INSDSeq_se

quence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="19">

<INSDSeq>

<INSDSeq_length>106</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..106</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q65">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>Homo sapiens</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>GQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPV

KAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS</INSDSeq_seq

uence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="20">

<INSDSeq>

<INSDSeq_length>107</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..107</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q66">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>Mus musculus</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>RADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQ

NGVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSSTSPIVKSFNRNEC</INSDSeq_se

quence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="21">

<INSDSeq>

<INSDSeq_length>106</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..106</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q67">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>Mus musculus</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>GQPKSSPSVTLFPPSSEELETNKATLVCTITDFYPGVVTVDWKVDGTPV

TQGMETTQPSKQSNNKYMASSYLTLTARAWERHSSYSCQVTHEGHTVEKSLSRADCS</INSDSeq_seq

uence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="22">

<INSDSeq>

<INSDSeq_length>120</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..120</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q68">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>QVTLRESGPALVKPTQTLTLTCTFSGFSLSTSGMSVGWIRQPPGKALEW

LADIWWDDKKDYNPSLKSRLTISKDTSKNQVVLKVTNMDPADTATYYCARSMITNWYFDVWGAGTTVTVS

S</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="23">

<INSDSeq>

<INSDSeq_length>106</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..106</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q69">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>DIQMTQSPSTLSASVGDRVTITCKCQLSVGYMHWYQQKPGKAPKLWIYD

TSKLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCFQGSGYPFTFGGGTKLEIK</INSDSeq_seq

uence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="24">

<INSDSeq>

<INSDSeq_length>126</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..126</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q70">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>Homo sapiens</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>QVQLVQSGAEVKKPGSSVMVSCQASGGLLEDYIINWVRQAPGQGPEWMG

GIIPVLGTVHYGPKFQGRVTITADESTDTAYMELSSLRSEDTAMYYCATETALVVSETYLPHYFDNWGQG

TLVTVSS</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="25">

<INSDSeq>

<INSDSeq_length>107</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..107</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q71">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>Homo sapiens</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>DIQMTQSPSSLSAAVGDRVTITCQASQDIVNYLNWYQQKPGKAPKLLIY

VASNLETGVPSRFSGSGSGTDFSLTISSLQPEDVATYYCQQYDNLPLTFGGGTKVEIK</INSDSeq_se

quence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="26">

<INSDSeq>

<INSDSeq_length>127</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..127</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q72">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>Homo sapiens</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>EVQLVESGGGLVRPGRSLRLSCTVSGFSFDDSAMSWVRQAPGKGLEWIS

FIKSKTYGGTKEYAASVKGRFTISRDDSKNIAYLQMNSLKTEDTAVYYCTRGAPYGGNSDYYYGLDVWGQ

GTTVTVSS</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="27">

<INSDSeq>

<INSDSeq_length>107</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..107</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q73">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>Homo sapiens</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>DIQMTQSPSSLSASVGDRVTITCRTSQDVRGALAWYQQKPGKAPKLLIF

DASSLETGVPSRFSGSGSGTVFTLTISSLQPEDFAAYYCQQFLDFPFTFGQGTRLEIK</INSDSeq_se

quence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="28">

<INSDSeq>

<INSDSeq_length>126</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..126</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q74">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>QVQLVQSGAEVKKPGSSVKVSCKASGGLLEDYIINWVRQAPGQGLEWMG

GIIPVLGTVHYGPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCATETAIVVTESYRPHFFDNWGQG

TLVTVSS</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="29">

<INSDSeq>

<INSDSeq_length>107</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..107</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q75">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>DIQMTQSPSSLSASVGDRVTITTCQASQDIVNYLNWYQQKPGKAPKLLIY

VASNLETGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQEFAGLALNFGGGTKVEIK</INSDSeq_se

quence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="30">

<INSDSeq>

<INSDSeq_length>126</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..126</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q76">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>QVQLVQSGAEVKKPGSSVMVSCQASGDPLQNYIINWVRQAPGQGPEWMG

AIIPALGSAHYGPKFQGRVTITADESTDTAYMELSSLRSEDTAMYYCATETAVIISESYLPHFFDNWGQG

TLVTVSS</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="31">

<INSDSeq>

<INSDSeq_length>107</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..107</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q77">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>DIQMTQSPSSLSAAVGDRVTITCQASQDIVNYLNWYQQKPGKAPKLLIY

VASNLETGVPSRFSGSGSGTDFSLTISSLQPEDVATYYCQEYDSLALAFGGGTKVEIK</INSDSeq_se

quence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="32">

<INSDSeq>

<INSDSeq_length>5</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..5</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q78">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>DYIIN</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="33">

<INSDSeq>

<INSDSeq_length>17</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..17</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q79">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>GIIPVLGTVHYGPKFQG</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="34">

<INSDSeq>

<INSDSeq_length>17</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..17</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q80">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>ETAIVVTESYRPHFFDN</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="35">

<INSDSeq>

<INSDSeq_length>11</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..11</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q81">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>QASQDIVNYLN</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="36">

<INSDSeq>

<INSDSeq_length>7</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..7</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q82">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>VASNLET</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="37">

<INSDSeq>

<INSDSeq_length>9</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..9</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q83">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>QEFAGLALN</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="38">

<INSDSeq>

<INSDSeq_length>5</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..5</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q84">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>NYIIN</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="39">

<INSDSeq>

<INSDSeq_length>17</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..17</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q85">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>AIIPALGSAHYGPKFQG</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="40">

<INSDSeq>

<INSDSeq_length>17</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..17</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q86">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>ETAVIISESYLPHFFDN</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="41">

<INSDSeq>

<INSDSeq_length>11</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..11</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q87">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>QASQDIVNYLN</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="42">

<INSDSeq>

<INSDSeq_length>7</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..7</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q88">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>VASNLET</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="43">

<INSDSeq>

<INSDSeq_length>9</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..9</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q89">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>QEYDSLALA</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="44">

<INSDSeq>

<INSDSeq_length>589</INSDSeq_length>

<INSDSeq_moltype>DNA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..589</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>genomic DNA</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q90">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>Escherichia coli</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>ttgagatccttttttctgcgcgtaatctgctgcttgcaaacaaaaaaa

ccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggct

tcagcagagcgcagataccaaatactgtccttctagtgtagccgtagttaggccaccacttcaagaactc

tgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcg

tgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggtt

cgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagctatgaga

aagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagag

cgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgac

ttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaa</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="45">

<INSDSeq>

<INSDSeq_length>455</INSDSeq_length>

<INSDSeq_moltype>DNA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..455</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>genomic DNA</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q91">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>f1 phage</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>cgcgccctgtagcggcgcattaagcgcggcgggtgtggtggttacgcgc

agcgtgaccgctacacttgccagcgccctagcgcccgctcctttcgctttcttcccttcctttctcgcca

cgttcgccggctttccccgtcaagctctaaatcggggggctccctttagggttccgatttagtgctttacg

gcacctcgaccccaaaaaacttgattagggtgatggttcacgtagtgggccatcgccctgatagacggtt

tttcgccctttgacgttggagtccacgttctttaatagtggactctcttgttccaaactggaacaacactca

accctatctcggtctattcttttgatttataagggattttgccgatttcggcctattggttaaaaaatga

gctgatttaacaaaaatttaacgcgaattttaacaaaatattaacgcttacaattt</INSDSeq_sequ

ence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="46">

<INSDSeq>

<INSDSeq_length>477</INSDSeq_length>

<INSDSeq_moltype>DNA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..477</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>genomic DNA</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q92">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>M13 phage</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>ggtggcggctccggttccggtgattttgattatgaaaaaatggcaaacg

ctaataagggggctatgaccgaaaatgccgatgaaaacgcgctacagtctgacgctaaaggcaaacttga

ttctgtcgctactgattacggtgctgctatcgatggtttcattggtgacgtttccggccttgctaatggt

aatggtgctactggtgattttgctggctctaattcccaaatggctcaagtcggtgacggtgataattcac

ctttaatgaataatttccgtcaatatttaccttccctccctcaatcggttgaatgtcgcccttttgtctt

tggcgctggtaaaccatatgaattttctattgattgtgacaaaataaacttattccgtggtgtctttgcg

tttcttttatatgttgccacctttatgtatgtattttctacgtttgctaacatactgcgtaataaggagt

cttgataa</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

</ST26SequenceListing>

<---

Claims (20)

1. An antibody to respiratory syncytial virus (RSV) comprising a heavy chain variable region comprising the heavy chain complementarity determining regions HCDR1, HCDR2 and HCDR3 and a light chain variable region comprising the light chain complementarity determining regions LCDR1, LCDR2 and LCDR3, wherein the antibody specifically binds to respiratory syncytial virus, and wherein the amino acid sequence of HCDR1 is presented in SEQ ID NO: 32, the amino acid sequence of HCDR2 is presented in SEQ ID NO: 33, the amino acid sequence of HCDR3 is presented in SEQ ID NO: 34, the amino acid sequence of LCDR1 is presented in SEQ ID NO: 35, the amino acid sequence of LCDR2 is presented in SEQ ID NO: 36 and the amino acid sequence of LCDR3 is presented in SEQ ID NO: 37; or the amino acid sequence of HCDR1 is presented in SEQ ID NO: 38, the amino acid sequence of HCDR2 is presented in SEQ ID NO: 39, the amino acid sequence of HCDR3 is presented in SEQ ID NO: 40, the amino acid sequence of LCDR1 is presented in SEQ ID NO: 35, the amino acid sequence of LCDR2 is presented in SEQ ID NO: 36 and the amino acid sequence of LCDR3 is presented in SEQ ID NO: 43; wherein the amino acid sequences of the complementarity-determining regions of the heavy chain and the complementarity-determining regions of the light chain are determined according to the Cabot nomenclature. 2. The antibody of claim 1, wherein the amino acid sequence of the variable region of the heavy chain of the antibody is presented in SEQ ID NO: 28 or 30. 3. The antibody of claim 1, wherein the amino acid sequence of the variable region of the light chain of the antibody is presented in SEQ ID NO: 29 or 31. 4. The antibody of claims 1-3, wherein the amino acid sequence of the variable region of the heavy chain of the antibody is at least 90% identical to the amino acid sequence defined in SEQ ID NO: 28 or 30, while the amino acid sequence of the variable region of the light chain of the antibody is at least 90% identical to the amino acid sequence defined in SEQ ID NO: 29 or 31. 5. The antibody of any one of claims 1 to 4, wherein the amino acid sequence of the variable region of the heavy chain of the antibody is presented in SEQ ID NO: 28, while the amino acid sequence of the variable region of the light chain of the antibody is presented in SEQ ID NO: 29; or the amino acid sequence of the variable region of the heavy chain of the antibody is presented in SEQ ID NO: 30, while the amino acid sequence of the variable region of the light chain of the antibody is presented in SEQ ID NO: 31. 6. The antibody of any one of claims 1 to 5, wherein the antibody is a neutralizing antibody. 7. The antibody of any one of claims 1 to 5, wherein the antibody is capable of binding to the F protein of human respiratory syncytial virus (RSV); wherein the antibody is preferably capable of binding a recombinant human RSV F protein, the sequence of which is presented in SEQ ID NO: 12 or 13. 8. The antibody of any one of claims 1 to 5, wherein the antibody is a Fab fragment, a whole antibody, an F(ab') ₂ fragment, or a single-chain Fv fragment (scFv fragment); wherein the antibody is preferably a Fab fragment. 9. The antibody of any one of claims 1 to 5, wherein the antibody is a monoclonal antibody. 10. The antibody of any one of claims 1 to 9, wherein the antibody comprises a heavy chain constant region of an IgG1 subtype, an IgG2 subtype, or an IgG4 subtype; wherein the heavy chain constant region preferably comprises an Fc region of an IgG1 subtype heavy chain constant region, wherein the amino acids at positions 252, 254, and 256 of the Fc region sequence are Y, T, and E, respectively, wherein the amino acid positions of the antibody constant region are determined according to EU numbering. 11. The antibody of any one of claims 1 to 9, wherein the antibody comprises a constant region of a light chain of the kappa subtype or the lambda subtype. 12. A nucleic acid molecule encoding an antibody according to any one of claims 1-11. 13. Use of an antibody according to any of claims 1-11 in the manufacture of a medicinal product for the prevention or treatment of a disease caused by RSV. 14. The use according to claim 13, wherein the disease caused by RSV is bronchiolitis or pneumonia. 15. A method for preventing or treating a disease caused by RSV, comprising the step of administering to a subject in need thereof an antibody according to any one of claims 1-11. 16. The method of claim 15, wherein the disease caused by RSV is bronchiolitis or pneumonia.