Three human papillomavirus antibodies and their use in preventing and treating HPV infection

Abstract

This disclosure provides three human papillomavirus (HPV) antibodies and their applications in the prevention and treatment of HPV infection. The antibodies are derived from memory B cells of vaccinated individuals. Humanized cross-neutralizing antibodies capable of neutralizing multiple HPV types were isolated. Sustained high concentrations of these neutralizing antibodies can neutralize viral particles continuously produced in the vaginal mucosa, preventing persistent viral infection of epithelial cells and helping the body accelerate the clearance of HPV. This provides a new approach for the development of antibody drugs for the clinical treatment of persistent HPV infection.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology and new medicine, specifically relating to three human papillomavirus antibodies and their application in the prevention and treatment of HPV infection. Background Technology

[0002] Among cancers caused by HPV infection, cervical cancer has received the most attention from researchers. HPV types 16 and 18 have been identified as the most common types associated with cervical cancer, causing more than 70% of cervical cancers and precancerous lesions (CIN). Cervical cancer is one of the most common cancers threatening women's health.

[0003] Human papillomavirus (HPV) is considered a necessary cause of cervical cancer development. Research data shows that the vast majority of cervical cancer patients are also infected with high-risk HPV. The majority of new cervical cancer cases worldwide originate in developing countries. Cervical cancer seriously affects the reproductive health of women in my country and globally. It takes approximately 3-5 years from high-risk HPV infection to the development of cervical intraepithelial neoplasia (CIN), and approximately 5-10 years from CIN to cervical cancer; a necessary condition for this is persistent infection with high-risk HPV.

[0004] Currently developed HPV preventative vaccines can provide some immune protection to vaccinated individuals. HPV virus-like particle (VLP) vaccines prevent HPV infection and induce high and durable titers of neutralizing antibodies to immunize against subsequent viral attacks. These neutralizing antibodies bind to and neutralize naturally occurring HPV virus particles. However, HPV preventative vaccines also have limitations. They only provide protection against the HPV types covered by the vaccine, offering poor cross-protection, and cannot treat individuals already infected with HPV. They are useless for those with persistent HPV infection. Currently, no drugs have been found that can directly and effectively clear high-risk HPV infections, causing significant anxiety and panic among women with persistent HPV infection, seriously impacting their mental and physical health—a problem urgently needing to be addressed clinically.

[0005] Treating individuals with persistent HPV infection presents a greater challenge in clinical practice than those with cervical lesions. Persistent HPV infection manifests as the continuous synthesis and release of new viral particles into the vaginal mucosal epithelial cells following initial infection. When the body's ability to clear these viral particles locally is weak, they cannot be eliminated promptly, leading to persistent infection. Therefore, effectively eliminating these viral particles remains a significant unresolved challenge. Summary of the Invention

[0006] To address at least one of the above-mentioned problems, this disclosure provides anti-human papillomavirus antibodies and their applications. Using the human papillomavirus antibodies provided by this disclosure, it is possible to develop drugs that offer new options for the clinical treatment of persistent HPV infection.

[0007] According to one aspect of this disclosure, an anti-human papillomavirus antibody or an antigen-binding fragment thereof is provided, the antibody or the antigen-binding fragment thereof comprising: (1) complementarity-determining regions of the following three light chain variable regions: LCDR1, having an amino acid sequence of LCDR1 contained in the light chain variable region as shown in any one of SEQ ID NO:19-21, or having one or more amino acid substitutions, deletions, additions, and / or having at least 80% sequence identity with the amino acid sequence of LCDR1 contained in the light chain variable region and retaining its human papillomavirus binding activity; LCDR2, having an amino acid sequence of LCDR2 contained in the light chain variable region as shown in any one of SEQ ID NO:19-21, or having one or more amino acid substitutions, deletions, additions, and / or having at least 80% sequence identity with the amino acid sequence of LCDR2 contained in the light chain variable region and retaining its human papillomavirus binding activity; LCDR3, having an amino acid sequence of LCDR2 contained in the light chain variable region as shown in ... The amino acid sequence of LCDR3 contained in the light chain variable region shown in any one of NO:19-21, or an amino acid sequence having one or more amino acid substitutions, deletions, additions, and / or having at least 80% sequence identity with the LCDR3 contained in the light chain variable region and retaining its human papillomavirus binding activity; and / or, (2) the complementarity-determining regions (HCDRs) of the following three heavy chain variable regions: HCDR1, having the amino acid sequence of HCDR1 contained in the heavy chain variable region shown in any one of SEQ ID NO:22-24, or having one or more amino acid substitutions, deletions, additions, and / or having at least 80% sequence identity with the HCDR1 contained in the heavy chain variable region and retaining its human papillomavirus binding activity; HCDR2, having the amino acid sequence of LCDR3 contained in the light chain variable region shown in SEQ ID NO:22-24, or an amino acid sequence having one or more amino acid substitutions, deletions, additions, and / or having at least 80% sequence identity with the LCDR3 contained in the heavy chain variable region and retaining its human papillomavirus binding activity; HCDR2, having the amino acid sequence of LCDR3 contained in the light chain variable region shown in SEQ ID NO:22-24, or having one or more amino acid substitutions, deletions, additions, and / or having at least 80% sequence identity with the LCDR3 contained in the heavy chain variable region and retaining its human papillomavirus binding activity; HCDR3, having the amino acid sequence of LCDR3 contained in the light chain variable region and having at least 80% sequence identity with the LCDR3 contained in the light ... The amino acid sequence of HCDR2 contained in the heavy chain variable region shown in any one of SEQ ID NO: 22 to 24, or an amino acid sequence having one or more amino acid substitutions, deletions, additions, and / or having at least 80% sequence identity with HCDR2 and retaining its human papillomavirus binding activity compared to the amino acid sequence of HCDR2 contained in the heavy chain variable region; HCDR3, having the amino acid sequence of HCDR3 contained in the heavy chain variable region shown in any one of SEQ ID NO: 22 to 24, or an amino acid sequence having one or more amino acid substitutions, deletions, additions, and / or having at least 80% sequence identity with HCDR3 and retaining its human papillomavirus binding activity compared to the amino acid sequence of HCDR3 contained in the heavy chain variable region.

[0008] In some implementations, each CDR is defined by any numbering system commonly used by those skilled in the art. Exemplary numbering systems include, but are not limited to, Kabat, AbM, Chothia, Contact, IMGT, or combinations thereof.

[0009] In some embodiments, the antibody or its antigen-binding fragment comprises:

[0010] (A1) The amino acid sequence of the three LCDRs contained in the light chain variable region as shown in SEQ ID NO:19, or an amino acid sequence having one or more amino acid substitutions, deletions, additions, and / or having at least 80% sequence identity with the three LCDRs contained in the light chain variable region and retaining its human papillomavirus binding activity compared to the amino acid sequence of any one of the three LCDRs contained in the light chain variable region; and / or, the amino acid sequence of the three HCDRs contained in the heavy chain variable region as shown in SEQ ID NO:22, or an amino acid sequence having one or more amino acid substitutions, deletions, additions, and / or having at least 80% sequence identity with the three HCDRs contained in the heavy chain variable region and retaining its human papillomavirus binding activity compared to the amino acid sequence of any one of the three HCDRs contained in the heavy chain variable region; or, (A2) the amino acid sequence of SEQ ID NO:19. The three LCDRs contained in the light chain variable region shown in NO:20, or an amino acid sequence having one or more amino acid substitutions, deletions, additions, and / or having at least 80% sequence identity with the three LCDRs contained in the light chain variable region and retaining its human papillomavirus binding activity compared to the amino acid sequence of any one of the three LCDRs contained in the light chain variable region; and / or, an amino acid sequence having one or more amino acid substitutions, deletions, additions, and / or having at least 80% sequence identity with the three LCDRs contained in the heavy chain variable region and retaining its human papillomavirus binding activity compared to the amino acid sequence of any one of the three LCDRs contained in the heavy chain variable region; or, (A3) an amino acid sequence as shown in SEQ ID NO:23. The amino acid sequence of the three LCDRs contained in the light chain variable region shown in NO:21, or an amino acid sequence having one or more amino acid substitutions, deletions, additions, and / or having at least 80% sequence identity with the three LCDRs contained in the light chain variable region and retaining its human papillomavirus binding activity; and / or the amino acid sequence of the three HCDRs contained in the heavy chain variable region shown in SEQ ID NO:24, or an amino acid sequence having one or more amino acid substitutions, deletions, additions, and / or having at least 80% sequence identity with the three HCDRs contained in the heavy chain variable region and retaining its human papillomavirus binding activity.

[0011] In some embodiments, the antibody or its antigen-binding fragment comprises: LCDR1, having an amino acid sequence as shown in any one of SEQ ID NO: 1 to 3, or having one or more amino acid substitutions, deletions, additions, and / or an amino acid sequence having at least 80% sequence identity and retaining its human papillomavirus binding activity compared to the LCDR1; and / or LCDR2, having an amino acid sequence as shown in any one of SEQ ID NO: 4 to 6, or having one or more amino acid substitutions, deletions, additions, and / or an amino acid sequence having at least 80% sequence identity and retaining its human papillomavirus binding activity compared to the LCDR2; and / or LCDR3, having an amino acid sequence as shown in any one of SEQ ID NO: 7 to 9, or having one or more amino acid substitutions, deletions, additions, and / or an amino acid sequence having at least 80% sequence identity and retaining its human papillomavirus binding activity compared to the LCDR3; and / or HCDR1, having an amino acid sequence as shown in SEQ ID NO: 1 to 3; and / or an amino acid sequence ... an amino acid sequence having one or more amino acid The amino acid sequence shown in any one of SEQ ID NO: 10 to 12, or an amino acid sequence having one or more amino acid substitutions, deletions, additions, and / or having at least 80% sequence identity with the amino acid sequence and retaining its human papillomavirus binding activity; and / or HCDR2, having an amino acid sequence shown in any one of SEQ ID NO: 13 to 15, or an amino acid sequence having one or more amino acid substitutions, deletions, additions, and / or having at least 80% sequence identity with the amino acid sequence and retaining its human papillomavirus binding activity; and / or HCDR3, having an amino acid sequence shown in any one of SEQ ID NO: 16 to 18, or an amino acid sequence having one or more amino acid substitutions, deletions, additions, and / or having at least 80% sequence identity with the amino acid sequence and retaining its human papillomavirus binding activity.

[0012] In some embodiments, the antibody or its antigen-binding fragment comprises: (B1) amino acid sequences such as LCDR1, LCDR2, and LCDR3 as shown in SEQ ID NO:1, SEQ ID NO:4, and SEQ ID NO:7, or an amino acid sequence having one or more amino acid substitutions, deletions, additions, and / or having at least 80% sequence identity with any of the amino acid sequences of LCDR1, LCDR2, and LCDR3 while retaining its human papillomavirus binding activity; and / or, amino acid sequences such as HCDR1, HCDR2, and HCDR3 as shown in SEQ ID NO:10, SEQ ID NO:13, and SEQ ID NO:16, or an amino acid sequence having one or more amino acid substitutions, deletions, additions, and / or having at least 80% sequence identity with any of the amino acid sequences of HCDR1, HCDR2, and HCDR3 while retaining its human papillomavirus binding activity; or, (B2) amino acid sequences such as SEQ ID NO:2, SEQ ID NO:5, and SEQ ID NO:6. The amino acid sequences shown in SEQ ID NO:8, such as LCDR1, LCDR2, and LCDR3, or those having one or more amino acid substitutions, deletions, additions, and / or having at least 80% sequence identity with the amino acid sequences of any one of LCDR1, LCDR2, and LCDR3 while retaining their human papillomavirus binding activity; and / or the amino acid sequences shown in SEQ ID NO:11, SEQ ID NO:14, and SEQ ID NO:17, such as HCDR1, HCDR2, and HCDR3, or those having one or more amino acid substitutions, deletions, additions, and / or having at least 80% sequence identity with the amino acid sequences of any one of HCDR1, HCDR2, and HCDR3 while retaining their human papillomavirus binding activity; or (B3) the amino acid sequences shown in SEQ ID NO:3, SEQ ID NO:6, and SEQ ID NO:8. The amino acid sequences shown in NO:9, such as LCDR1, LCDR2, and LCDR3, or those having one or more amino acid substitutions, deletions, additions, and / or having at least 80% sequence identity with the amino acid sequences of any one of LCDR1, LCDR2, and LCDR3 while retaining their human papillomavirus binding activity, and / or those having amino acid sequences such as HCDR1, HCDR2, and HCDR3 shown in SEQ ID NO:12, SEQ ID NO:15, and SEQ ID NO:18, such as HCDR1, HCDR2, and HCDR3, or those having one or more amino acid substitutions, deletions, additions, and / or having at least 80% sequence identity with the amino acid sequences of any one of HCDR1, HCDR2, and HCDR3 while retaining their human papillomavirus binding activity.

[0013] In some embodiments, the antibody or its antigen-binding fragment comprises: a light chain variable region comprising an amino acid sequence as shown in any one of SEQ ID NO: 19-21, or an amino acid sequence having one or more amino acid substitutions, deletions, or additions compared to the antibody, and / or an amino acid sequence having at least 80% sequence identity with the antibody and retaining its human papillomavirus binding activity; and / or a heavy chain variable region comprising an amino acid sequence as shown in any one of SEQ ID NO: 22-24, or an amino acid sequence having one or more amino acid substitutions, deletions, or additions compared to the heavy chain, and / or an amino acid sequence having at least 80% sequence identity with the heavy chain and retaining its human papillomavirus binding activity.

[0014] In some embodiments, the antibody or its antigen-binding fragment comprises:

[0015] (C1) A light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 19, or an amino acid sequence having one or more amino acid substitutions, deletions, additions, and / or having at least 80% sequence identity with the amino acid sequence and retaining its human papillomavirus binding activity; and / or a heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 22, or an amino acid sequence having one or more amino acid substitutions, deletions, additions, and / or having at least 80% sequence identity with the amino acid sequence and retaining its human papillomavirus binding activity; or (C2) a light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 20, or an amino acid sequence having one or more amino acid substitutions, deletions, additions, and / or having at least 80% sequence identity with the amino acid sequence and retaining its human papillomavirus binding activity; and / or a heavy ... The amino acid sequence shown in SEQ ID NO: 23, or an amino acid sequence having one or more amino acid substitutions, deletions, additions, and / or having at least 80% sequence identity with it and retaining its human papillomavirus binding activity; or (C3) a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 21, or an amino acid sequence having one or more amino acid substitutions, deletions, additions, and / or having at least 80% sequence identity with it and retaining its human papillomavirus binding activity; and / or a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 24, or an amino acid sequence having one or more amino acid substitutions, deletions, additions, and / or having at least 80% sequence identity with it and retaining its human papillomavirus binding activity.

[0016] In some embodiments, the antibody or antigen-binding fragment may be, but is not limited to, IgA, IgD, IgE, IgG, or IgM.

[0017] In some embodiments, the antibody or antigen-binding fragment includes, but is not limited to, scFv, Fab, Fab', (Fab')2, Fv fragment, dsFv, bispecific antibody, and multispecific antibody;

[0018] In some embodiments, the antibody or antigen-binding fragment is used to neutralize human papillomavirus (HPV).

[0019] In some embodiments, the human papillomavirus includes HPV6, HPV11, HPV16, HPV18, HPV31, HPV33, HPV45, HPV52, or HPV58.

[0020] In some embodiments, the antibody or antigen-binding fragment includes, but is not limited to, murine antibodies, chimeric antibodies, humanized antibodies, and fully human antibodies.

[0021] In this disclosure, the antigen-binding fragment typically includes at least a portion of the antigen-binding region or variable region of the antibody, such as one or more CDRs. The antibody fragment retains at least some of the antibody's binding specificity.

[0022] According to another aspect of this disclosure, a nucleic acid molecule is provided that encodes the antibody or an antigen-binding fragment thereof.

[0023] According to another aspect of this disclosure, an expression vector is provided that comprises the nucleic acid molecule and / or the expression vector.

[0024] In some embodiments, the expression vector includes prokaryotic expression vectors and eukaryotic expression vectors.

[0025] In some embodiments, the eukaryotic expression vector includes yeast expression vectors, mammalian expression vectors, insect expression vectors, etc.

[0026] According to another aspect of this disclosure, a host cell is provided that contains the nucleic acid molecule and / or the expression vector.

[0027] In some embodiments, the host cell is selected from prokaryotic cells and eukaryotic cells.

[0028] In some embodiments, the prokaryotic cells include bacterial cells.

[0029] In some embodiments, the bacterial cells include Escherichia coli and Streptomyces;

[0030] In some embodiments, the eukaryotic cells include yeast cells, mammalian cells, insect cells, etc.

[0031] In some embodiments, the mammal is selected from humans, monkeys, mice, rats, hamsters, goats, sheep, cattle, pigs, dogs, or cats.

[0032] In some embodiments, the mammalian cells include CHO cells, 293 cells, Vero cells, BHK cells, NSO cells, SP2 / 0 cells, YO myeloma cells, P3X63 mouse myeloma cells, PER cells, PER.C6 cells, or hybridoma cells.

[0033] According to another aspect of this disclosure, a chimeric antigen receptor is provided, comprising an extracellular antigen-binding domain, a transmembrane domain, and an intracellular signal transduction domain, wherein the extracellular antigen-binding domain comprises the antibody or an antigen-binding fragment thereof.

[0034] According to another aspect of this disclosure, a modified immune cell is provided that includes the chimeric antigen receptor described above.

[0035] According to another aspect of this disclosure, an antibody-drug conjugate is provided, comprising: the antibody or an antigen-binding fragment thereof; and a drug covalently linked to the antibody or the antigen-binding fragment thereof.

[0036] According to another aspect of this disclosure, a detection kit is provided, the kit comprising the antibody or an antigen-binding fragment thereof.

[0037] According to another aspect of this disclosure, a composition is provided comprising the antibody or an antigen-binding fragment thereof, and a pharmaceutically acceptable carrier.

[0038] In some embodiments, the antibody or its antigen-binding fragment comprises one or more of (A1), (A2), and (A3).

[0039] In some embodiments, the antibody or its antigen-binding fragment comprises: (A1), (A2), and (A3).

[0040] According to another aspect of this disclosure, the use of the antibody or its antigen-binding fragment, the nucleic acid molecule, the expression vector, the host cell, the chimeric antigen receptor, the modified immune cell, the antibody-drug conjugate, the detection kit, and / or the composition in drug research, drug preparation, non-clinical research, clinical research, and clinical testing is provided.

[0041] According to another aspect of this disclosure, the use of the antibody or its antigen-binding fragment, the nucleic acid molecule, the expression vector, the host cell, the chimeric antigen receptor, the modified immune cell, the antibody-drug conjugate, the detection kit, and / or the composition in the preparation of products for the diagnosis, prevention, and / or treatment of human papillomavirus (HPV) infection is provided.

[0042] According to another aspect of this disclosure, a method for detecting human papillomavirus is provided, the method comprising the step of using the said antibody or an antigen-binding fragment thereof for detection.

[0043] According to another aspect of this disclosure, a method for preventing or treating human papillomavirus infection is provided, comprising administering to a subject a therapeutically effective amount of the antibody or its antigen-binding fragment or the composition thereof.

[0044] Beneficial effects:

[0045] This disclosure discloses the isolation of high and medium levels of anti-human papillomavirus (HPV) antibodies from the serum of volunteers vaccinated against HPV. These three antibodies, derived from memory B cells of the vaccinated population, yielded humanized cross-neutralizing antibodies capable of neutralizing multiple HPV types, covering several major HPV types. The sustained high concentrations of neutralizing antibodies can neutralize viral particles continuously produced in the vaginal mucosa, preventing persistent viral infection of epithelial cells and helping the body accelerate the clearance of HPV. This provides a new approach for the clinical treatment of persistent HPV infection and the development of broad-spectrum or HPV-type-specific antibody drugs. Attached Figure Description

[0046] Figure 1 The technical routes for antibody screening, preparation and detection described in Examples 1 to 4 are shown.

[0047] Figure 2 An orthogonal view of the H419 Fab fragment and the L1 pentamer complex is shown. Blue and orange represent the light and heavy chains of the antibody, while the other five colors represent the pentamer of the HPV L1 protein.

[0048] Figure 3 An orthogonal view of the H421 Fab fragment and the L1 pentamer complex is shown. Blue and orange represent the light and heavy chains of the antibody, while the other five colors represent the pentamer of the HPV L1 protein.

[0049] Figure 4 An orthogonal view of the H7 Fab fragment and L1 pentamer complex is shown. Blue and orange represent the light and heavy chains of the antibody, while the other five colors represent the pentamer of the HPV L1 protein. Detailed Implementation

[0050] The cervix, located between the vagina and uterus, is characterized by a single layer of columnar secretory epithelium. HPV infects basal epithelial cells through epithelial abrasion or wounds, and can also enter epithelial cells through cells at the junction of the single layer of squamous cells inside and outside the cervix. HPV infection of epithelial cells induces cervical dysplasia and cervical intraepithelial neoplasia (CIN), which can further develop into cervical cancer due to persistent infection with high-risk HPV types. After viral infection, early HPV genes (E1, E2, E4, E5, E6, and E7) are expressed during the reproductive cycle. In the upper layer of epithelial tissue, the viral genome replicates, and late HPV genes (L1 and L2) and E4 are expressed, assembling progeny HPV viral particles. Shed viral particles repeat the viral life cycle, continuously infecting the remaining epithelial tissue cells. Persistent infection with high-risk HPV types for more than one year increases the risk of developing advanced squamous intraepithelial lesions or potentially invasive cancer. Currently, there are no drugs that can effectively clear high-risk HPV infection, and persistent high-risk HPV infection causes great anxiety and panic among women.

[0051] Based on the process of HPV virus invading epithelial cells, it is known that a sustained high concentration of neutralizing antibodies can neutralize the viral particles continuously produced in the vaginal mucosa, preventing the virus from continuously infecting epithelial cells and helping the body accelerate the clearance of HPV virus. Studies have shown that the initial binding of the human papillomavirus capsid to host cells is mainly through the interaction between the viral capsid protein L1 and heparan sulfate proteoglycan (HSPG) on the cell surface.

[0052] Therefore, this disclosure isolates high and high levels of anti-HPV L1 protein antibodies from the serum of volunteers vaccinated against HPV. These three antibodies (H419, H7, and H421) are derived from memory B cells of vaccinated individuals and have not undergone any sequence optimization; their sequences are shown in Table 1. The binding affinity and neutralization levels of these antibodies against nine HPV types were also tested. Antibody drugs prepared from these neutralizing antibodies can directly act on the vaginal mucosa, neutralizing viral particles shed from infected vaginal areas, preventing newly generated HPV viruses from persistently infecting vaginal epithelial cells, and assisting the body in clearing HPV viruses, providing a new approach for the clinical treatment of persistent HPV infection.

[0053] Table 1. Amino acid sequences of the CDR regions of the three antibodies

[0054]

[0055]

[0056] Note: The CDR regions described in Table 1 are defined by IMGT rules.

[0057] definition

[0058] Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly used in the field to which this invention pertains. For the purposes of interpreting this specification, the following definitions will apply, and where appropriate, terms used in the singular will also include the plural forms, and vice versa.

[0059] Unless the context clearly indicates otherwise, the terms “a” and “an” as used herein include plural references.

[0060] The term "about" as used herein is as understood by one of ordinary skill in the art and varies within a certain range depending on the context in which it is used. If one of ordinary skill in the art is unfamiliar with the use of this term in the context in which it is used, "about" will mean a particular value plus or minus 10%.

[0061] As used herein, the term "specific binding" refers to binding selectivity for an antigen, which can be distinguished from unwanted or nonspecific binding. The ability of an antigen-binding molecule to bind to a specific antigen can be measured using techniques familiar to those skilled in the art, such as surface plasmon resonance (SPR), enzyme-linked immunosorbent assay (ELISA), or conventional binding assays.

[0062] As used herein, the term "antibody" encompasses a wide range of antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies, monospecific and multispecific antibodies (e.g., bispecific or trispecific antibodies), single-chain molecules, and antibody fragments, as long as they exhibit the desired antigen-binding activity. Each heavy chain may include a heavy chain variable region (VH) and a heavy chain constant region (CH). Each light chain may include a light chain variable region (VL) and a light chain constant region (CL).

[0063] As used herein, the term "antigen-binding fragment" refers to an antibody fragment that retains the ability to specifically bind to an antigen bound by a full-length antibody, such as a fragment retaining one or more CDR regions. Examples of antigen-binding fragments include, but are not limited to, Fab, Fab', F(ab')2, and Fv fragments; biantibodies; linear antibodies; single-chain antibody molecules, such as single-chain Fv (ScFv); nanobodies, etc.

[0064] The term "Fab fragment" as used in this article refers to the region of an antibody structure that can bind to an antigen. A Fab fragment consists of a complete light chain and a partial heavy chain structure, linked by a disulfide bond, and is relatively small in size. Fab fragments can be obtained by enzymatic digestion of full-length antibodies. For example, human immunoglobulin G (lgG) can be degraded into two Fab fragments and one Fc fragment by papain; or lgG can be degraded into an F(ab')2 fragment and a pFc' fragment by pepsin. The F(ab')2 fragment can be further reduced to form two Fab' fragments. Fab fragments can also be prepared by expression in prokaryotic systems (such as the E. coli system) and mammalian cell systems. The E. coli expression system has the advantages of low production cost and fast production speed, but it is prone to inclusion body formation, making subsequent purification and refolding more troublesome, and the refolded protein has very low or even no activity. Fab fragment expression in mammalian cells allows for successful disulfide bond formation, more closely resembling the structure of the natural Fab fragment, and exhibits higher activity.

[0065] The term "complementarity-determining region (CDR)" refers to a site in the variable region of an antibody that confers specificity for binding to an antigen. Those skilled in the art will understand that, unless otherwise specified, the terms "CDR" and "complementarity-determining region" for a given antibody or its region (e.g., the variable region) should be understood to encompass "CDR" and "complementarity-determining region" as defined by any of the methods known in the art. It is well known to those skilled in the art that the CDR of an antibody can be defined in various ways, such as the Kabat definition rule based on sequence variability, the Chothia definition rule based on the location of structural loop regions, the Martin definition rule based on sequence and frame regions, the IMGT definition rule based on germline V gene amino acid sequence alignment, and reference tools for antibody humanization design based on CDR transplantation, etc. Those skilled in the art will understand that, unless otherwise specified, the terms "CDR" and "complementarity-determining region" for a given antibody or its region (e.g., the variable region) should be understood to encompass "CDR" as defined by any of the known schemes described above, and the amino acid sequences corresponding to the defined CDR rules should also fall within the scope of protection of this invention.

[0066] As used in this article, the term "chimeric antibody" refers to an antibody that combines antibody fragments from different species. Specifically, for example, a monoclonal antibody from one species (e.g., mouse) whose Fc constant region is replaced by an Fc constant region from another species (e.g., human) via DNA recombination technology.

[0067] As used herein, the term "humanized antibody" refers to an antibody that contains the human immunoglobulin framework region and one or more core-residue junctions (CDRs) derived from non-human (e.g., mouse, rat, rabbit, or synthetic) immunoglobulins. Except for the CDRs, all other parts of a humanized antibody are substantially identical to the corresponding parts of the native human immunoglobulin sequence.

[0068] As used herein, the term "fully human antibody" is intended to include antibodies possessing variable and constant regions derived from human immunoglobulin sequences. As used herein, the term "fully human antibody" refers to an antibody whose protein molecule is virtually non-immunogenic in humans, with only minor sequence variations or alterations relative to human natural immunoglobulins, encompassing almost all parts of the protein molecule (e.g., CDR, FR, CL, HC domains (e.g., CH1, CH2, CH3), hinges, VL, VH). Therefore, fully human antibodies differ from chimeric or humanized antibodies. It should be noted that fully human antibodies can be produced by human cells, non-human animals, or prokaryotic or eukaryotic cells capable of expressing functionally rearranged human immunoglobulin (e.g., heavy chain and / or light chain) genes.

[0069] As used herein, the term "composition" refers to any composition comprising at least one bioactive pharmaceutical agent, such as the fusion protein. When used herein, the term "composition" also refers to a composition comprising an active pharmaceutical ingredient to be delivered to a subject to achieve, for example, a therapeutic, preventative, diagnostic, inhibitory, or prognostic effect. The compositions of the present invention can be administered as food or health products, such as nutritional supplements. Generally, the compositions of the present invention are intended for the treatment of humans, although they can be used to treat animals, including monogastric mammals such as poultry, pigs, cats, dogs, horses, or rabbits. The compositions of the present invention can be used to enhance the growth and performance of animals. If administered to animals, oral feeding can be used.

[0070] As used in this disclosure, the term "pharmaceutically acceptable carrier" refers to a substance suitable for human and / or animal use without excessive adverse side effects (such as toxicity, irritation, and allergic reactions), i.e., a reasonable benefit / risk ratio. Examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as methylcellulose, ethylcellulose, hydroxypropyl methylcellulose, sodium carboxymethylcellulose, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (such as Tween), wetting agents (such as sodium lauryl sulfate), buffers, chelating agents, thickeners, pH adjusters, transdermal penetration enhancers, colorants, flavoring agents, stabilizers, antioxidants, preservatives, antibacterial agents, pyrogen-free water, etc.

[0071] As used in this article, the term "nucleic acid molecule" refers to any one or more nucleic acid segments present in a polynucleotide, such as DNA or RNA fragments.

[0072] As used herein, "percentage homology," "% homology," or "% sequence identity" describes the degree of similarity between two nucleotide sequences or two amino acid sequences and has the same meaning as "percentage identity." The percentage homology of two sequences can be calculated by dividing the number of identical residue positions by the total length of the aligned sequences and then multiplying by 100%. Methods and tools for aligning two amino acid or nucleotide sequences are well known in the art, such as the BLAST kit available on the NCBI website. The phrase "at least 85% sequence identity" as used herein refers to having at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with that sequence.

[0073] As used herein, the term "expression vector" refers to a DNA molecule to which a specific gene, operatively linked, is introduced into a target cell and directed to express it. Expression vectors include vectors that function as self-replicating nucleic acid structures and vectors that are incorporated into the genome of the host cell into which they have been introduced. Expression vectors can perform transcription of large amounts of stable mRNA. Once the expression vector is within the target cell, cellular transcription and / or translation mechanisms generate the ribonucleic acid molecule or protein encoded by that gene.

[0074] The term "host cell" as used herein is used interchangeably and refers to a cell in which exogenous nucleic acids have been introduced, including the progeny of such cells. Host cells include primary transformed cells and their derived progeny. The nucleic acids of the progeny may be completely identical to or not completely identical to those of the parent cells, and may contain mutations. Host cells include cultured cells, such as cultured mammalian cells, such as CHO cells, 293 cells, Vero cells, BHK cells, NS0 cells, SP2 / 0 cells, YO myeloma cells, P3X63 mouse myeloma cells, PER cells, PER.C6 cells, or hybridoma cells, yeast cells, insect cells, and plant cells. The host cells disclosed herein also include cells contained within transgenic animals, transgenic plants, or cultured plant or animal tissues.

[0075] The composition can be in any dosage form, including, but not limited to, tablets, pills, powders, granules, capsules, suspensions, solutions, emulsions, syrups, injections, sprays, aerosols, powder sprays, lotions, liniments, ointments, plasters, pastes, patches, eye drops, nasal drops, suppositories, etc.

[0076] The compositions disclosed herein can be administered in pharmaceutically effective amounts. "Pharmaceutically effective amount" refers to an amount sufficient to treat a disease and a reasonable benefit / risk ratio suitable for any medical treatment. The effective dose level of the composition can be determined based on the subject's type, disease severity, age and sex, drug activity, drug sensitivity, time of administration, route of administration, excretion rate, duration of treatment, drugs used in combination with the composition, and other factors known in the medical field. The compositions disclosed herein can be used alone or in combination with other therapeutic agents, and can be administered sequentially or simultaneously with conventional therapeutic agents. The compositions can be administered in one or more dosage forms.

[0077] As used herein, the term "treatment" for a subject's disease or "treatment" for a subject who has or is suspected of having a disease refers to administering medication to the subject, such as one or more agents, to reduce or prevent the worsening of at least one symptom of the disease. Therefore, in some embodiments, "treatment" specifically refers to delaying progression, accelerating remission, inducing remission, increasing remission, accelerating recovery, increasing the efficacy of alternative therapies, or reducing resistance to alternative therapies, or combinations thereof.

[0078] The term "diagnosis" as used herein refers not only to "judgment" but also includes "determination," "differentiation," "identification," "examination," "detection," and "prediction." In some embodiments, the diagnosis includes differentiating or assisting in differentiating lesions, determining the degree of disease progression, or predicting the risk of disease development, treatment effectiveness, prognosis, and recurrence risk.

[0079] The term “prevention” as used herein is recognized in the art and is well known in the art when used in connection with conditions such as local recurrence, and includes administration that reduces or delays the onset of medical condition symptoms in a subject relative to a subject who has not received the composition.

[0080] The term “EC50” as used in this article refers to the half-maximal concentration, which is the antibody concentration that can produce a 50% maximum effect (in this article, the antibody’s ability to bind to the HPVVLP protein).

[0081] As used herein, the term "IC50" refers to the half-maximal inhibitory concentration, i.e., the antibody concentration that produces a 50% maximum inhibitory effect (in this document, the ability to inhibit cell activity). Examples and figures are provided below to aid in understanding the invention. However, it should be understood that these examples and figures are for illustrative purposes only and do not constitute any limitation. The actual scope of protection of the invention is set forth in the claims. It should be noted that various modifications and improvements made by those skilled in the art based on this inventive concept are within the scope of protection of this invention. In the following description, descriptions of well-known structures and techniques are omitted to avoid unnecessarily obscuring the concepts of this disclosure. Such structures and techniques have also been described in many publications. Furthermore, reagents used in the examples, unless otherwise specified, are all commercially available products.

[0082] Example

[0083] The technical routes for antibody screening, preparation, and detection described in Examples 1-4 are as follows: Figure 1 .

[0084] Example 1:

[0085] 1) Volunteer blood sample collection

[0086] One month after receiving the HPV vaccine, volunteers had 100 mL of blood collected. Volunteers were between 18 and 38 years old and had no history of vaccine allergies. The experimental protocol and ethical review were approved by the Ethics Review Committee of Peking Union Medical College Hospital, and all participants signed written informed consent forms.

[0087] 2) PBMC cell isolation

[0088] Volunteer blood was subjected to density gradient centrifugation using 50 mL centrifuge tubes containing Ficoll-Hypaque (Cytiva) lymphocyte separation medium at the bottom. After washing multiple times with Hanks buffer (Solarbio), the separated PBMCs were placed in 90% heat-inactivated FBS supplemented with 10% dimethyl sulfoxide (DMSO) and stored in liquid nitrogen until thawed for use in experiments.

[0089] 3) Flow cytometry for sorting mononuclear B cells

[0090] Frozen PBMCs were rapidly thawed in a 37°C water bath, then added to FACS buffer (PBS containing 2% fetal bovine serum (PAN Biotech)), centrifuged at 300g for 8 min, and the supernatant was discarded. The cells were resuspended in FACS buffer. The cell suspension was filtered through a 40μm cell sieve and counted. Fluorescent staining was performed, with approximately 1 × 10⁶ cells per negative and single-stain tube.6 One cell was transferred to a 1.5 mL Eppendorf tube; the remaining cells were used as sample tubes for fluorescent staining according to the experimental purpose. The staining solution included Percp-Cy5.5 anti-human CD3 (BioLegend), APC anti-human CD27 (BioLegend), Pacific Blue anti-human CD19 (BioLegend), PE anti-human IgG (BioLegend), PerCP / Cy5.5 anti-human IgM (BioLegend), and AF488-labeled VLP proteins of various HPV types. The cells were blocked at 4°C in the dark for 30 min. Afterwards, the cells were washed twice with 500 μL of FACS buffer and placed on ice for sorting. Using a BD FACSAria III flow cytometer (BD Biosciences), single memory B cells bound to AF488-labeled protein were sorted into 96-well PCR plates, followed by antibody sequencing and clonal expression of B cell monoclonal antibodies.

[0091] 4) Cloning, expression and purification of antibodies

[0092] Nucleic acid sequences encoding the variable regions of naturally paired antibody heavy and light chains were synthesized and subcloned into the pcDNA3.1 vector for expression in HEK293F cells. Recombinant plasmids encoding each antibody heavy and light chain were transiently co-transfected into HEK293F cells and cultured at 37°C for 7 days at 8% CO2 to achieve antibody expression. After 7 days of expression, the cell suspension was collected and centrifuged at 3400 rpm, 4°C for 40 min, and the cell culture supernatant was collected. The supernatant was filtered using a 0.45 μM filter cup and concentrated using a membrane with a 100 kDa cutoff, with the medium replaced more than 100 times with PBS. The intact antibody containing the Fc fragment was attached to the medium using a protein A affinity chromatography gravity column (Tiandi Renhe Company), and then eluted with IgG Elution Buffer (Pierce). After concentration and replacement with PBS, the intact antibody protein was obtained.

[0093] Example 2: ELISA binding assay of humanized antibody to HPV type 9 VLP protein

[0094] HPV VLP proteins (GenBank accessions: HPV6 AAC80442.1; HPV11 AAA46935.1; HPV16 ANA05496; HPV18 AAQ92369; HPV31 P17388; HPV33 AMY16565; HPV45 P36741; HPV52 AML80965; HPV58 AFS33402) were diluted to 2 μg / mL with PBS for antigen coating. 100 μL of the coating solution was added to each well of an ELISA plate and incubated overnight at 4°C. The coated plates were then washed three times with PBST (PBS + 0.05% Tween 20), 300 μL each time. Then, add 300 μL of 2% BSA to each well for blocking and incubate at room temperature for 2 hours. Wash the blocked plate three times with PBST. Dilute the original 1 mg / mL antibody with PBS (the first well is diluted 1000-fold) in eight 1:3 gradients and add 100 μL to each well of the corresponding ELISA plate. Incubate at 37°C for 30 minutes. Then, add 100 μL of 1:5000 diluted secondary antibody (HRP-conjugated anti-human IgG (Sigma-Aldrich)) to each well and incubate at 37°C for 30 minutes. Wash the plate five times with PBST. Add 100 μL of TMB chromogenic solution to each well and incubate at room temperature in the dark for 15 minutes. The reaction was then terminated by adding 50 μL of 2M H2SO4 solution. The plate was read using a microplate reader, and the absorbance value was read at 450 nm (OD value). Candidate antibodies H419, H7, and H421 with high binding capacity were screened out (the amino acid sequences of their heavy chain variable region, light chain variable region, and CDR are shown in Table 1, and the DNA sequences of their light chain variable region and heavy chain variable region are shown in Table 2). The antibody concentration at which the absorbance value reached 50% of the maximum value was recorded as the EC50 of the antibody. 50 Table 3 shows the EC values ​​of these three antibodies. 50 The values ​​show that the selected HPV antibodies have good cross-binding ability among HPV types 18 / 45, 16 / 31 / 52, and 6 / 11 / 33.

[0095] Table 2. DNA sequences of the light chain variable region and heavy chain variable region of the three antibodies

[0096]

[0097]

[0098] Table 3. Binding ability of HPV antibodies to various VLP proteins

[0099]

[0100]

[0101] Example 3: Detection of antibody neutralizing ability using a pseudovirus neutralization experiment

[0102] The antibody concentration was adjusted to 1 mg / mL. 250 μL of sterile water was added to the wells of a 96-well plate for edge sealing to reduce experimental error. The antibody was serially diluted seven times (4-fold) starting from 30-fold. An equal volume of HPV pseudovirus (provided by the China National Institutes for Food and Drug Control) was added to the antibody serial dilutions in the 96-well plate, and the plate was incubated at 37°C for 1 hour. After incubation, 100 μL of digested HEK 293T cells was added to each well. The plate was then incubated at 37°C in a 5% CO2 cell culture incubator for 72 hours. The plate was then analyzed using a Cytation5 microplate reader. The inhibition rate (%) was calculated as: 1 - (sample detection value - cell control value) / (virus control value - cell control value) × 100 (%). The antibody concentration corresponding to a 50% inhibition rate was taken as the IC50. 50 The IC50 of the candidate antibody was determined by in vitro pseudovirus neutralization assay. 50 As shown in Table 4, these HPV antibodies exhibit good cross-neutralizing activity against HPV types 18 / 45, 16 / 31 / 52 / 58, and 6 / 11 / 33.

[0103] Table 4 Neutralizing activity of HPV antibodies against various types of pseudoviruses

[0104]

[0105] Meanwhile, in order to ensure that the antibody drug can significantly neutralize all nine types of HPV, we mixed three antibodies to test the neutralizing antibody titers against HPV types 6, 11, 16, 18, 31, 33, 45, 52, and 58 pseudoviruses. As shown in Table 5, the mixed antibody showed good neutralizing effect against all nine types of HPV pseudoviruses.

[0106] Table 5. Neutralization levels of cross-neutralizing antibodies against three groups of mixed HPV antibodies.

[0107]

[0108] Example 4: Cryo-electron microscopy sample preparation and data collection

[0109] Because the HPVVLP protein morphology is not uniform and is easily deformed under electron microscopy, which is not conducive to subsequent structural analysis, we depolymerized the VLP protein into a uniform L1 pentamer using the reducing agent dithiothreitol (DTT). Specifically, the HPVVLP protein was repeatedly concentrated and buffered in a centrifugal ultrafiltration tube using a depolymerization buffer containing 1 mM DTT until a dilution ratio of 1:1000 was achieved.

[0110] 0.8 mg / mL of HPV L1 pentamer protein was mixed with the purified HPV antibody Fab fragment at a molar ratio of 1:1.2 and incubated on ice for 30 min for sample preparation. A 300-mesh gold grid (C-flat, CF-1.2 / 1.3-3Au-50, USA) covered with a discontinuous carbon film was used. First, the grid was subjected to glow discharge. The grid was placed in a Solarus plasma cleaner (Gatan, USA) with a mixed H2 and O2 gas mode, a discharge power of 25 W, and a discharge time of 25 s to complete the hydrophilication treatment of the grid. The Vitrobot cryogenic sample preparation instrument was adjusted to the following parameters: temperature 22℃, ambient humidity 100%, filter paper clamping time 6 s, and clamping force 0. Then, the treated gold grid was held with sample preparation tweezers, and 3 μL of the premixed sample was adsorbed onto the grid. Lightly press the foot control panel to start the preset program. After the filter paper is clamped onto the carrier mesh, it is quickly immersed in the prepared liquid ethane to prepare the sample.

[0111] Cryo-electron microscopy sample data were then collected using a FEI Titan Krios 300kV cryo-electron microscope, and data analysis was performed using software such as Relion 3.0 and cryoSPARC. Sample structures are shown below. Figures 2-4 As shown in the figure, the structural studies on the antibody Fab fragment and the L1 capsid protein that makes up the surface of HPV viral particles revealed the structural basis for their binding and neutralizing abilities. Most of the cross-neutralizing antibodies we selected bound to relatively conserved amino acid sites at the apex of the corresponding HPV type L1 pentamer. The antibody complex structure further validated previous experimental results, providing structural support for the use of HPV neutralizing antibody drugs to neutralize viral particles shed from infected vaginal sites.

[0112] The technical solutions of the present invention are not limited to the specific embodiments described above. Any technical modifications made in accordance with the technical solutions of the present invention fall within the protection scope of the present invention.

Claims

1. An anti-human papillomavirus antibody or an antigen-binding fragment thereof, said antibody or antigen-binding fragment comprising: LCDR1, LCDR2, and LCDR3 contained in the light chain variable region as shown in SEQ ID NO: 20; and HCDR1, HCDR2, and HCDR3 contained in the heavy chain variable region as shown in SEQ ID NO: 23; in, The LCDR1, LCDR2, LCDR3, HCDR1, HCDR2, and HCDR3 are defined based on the same Kabat, AbM, Chothia, Contact, or IMGT numbering system.

2. The antibody or its antigen-binding fragment according to claim 1, characterized in that, The antibody or its antigen-binding fragment comprises: LCDR1, LCDR2 and LCDR3 with amino acid sequences as shown in SEQ ID NO: 2, SEQ ID NO: 5 and SEQ ID NO: 8; and HCDR1, HCDR2 and HCDR3 with amino acid sequences as shown in SEQ ID NO: 11, SEQ ID NO: 14 and SEQ ID NO:

17.

3. The antibody or its antigen-binding fragment according to claim 1, characterized in that, The antibody or its antigen-binding fragment comprises: a light chain variable region containing an amino acid sequence as shown in SEQ ID NO: 20; and a heavy chain variable region containing an amino acid sequence as shown in SEQ ID NO:

23.

4. The antibody or its antigen-binding fragment according to claim 1, characterized in that, The antibody or antigen-binding fragment includes IgA, IgD, IgE, IgG, or IgM types.

5. The antibody or its antigen-binding fragment according to claim 1, characterized in that, The antibody or antigen-binding fragments include scFv, Fab, Fab', (Fab')2, Fv fragments, and dsFv.

6. The antibody or its antigen-binding fragment according to claim 1, characterized in that, The antibody or antigen-binding fragment is used to neutralize human papillomavirus, wherein the human papillomavirus is selected from HPV16, HPV18, HPV31, HPV33, HPV52 or HPV58.

7. The antibody or its antigen-binding fragment according to claim 1, characterized in that, The antibody or antigen-binding fragment includes murine antibodies, chimeric antibodies, humanized antibodies, or fully human antibodies.

8. A nucleic acid molecule encoding an antibody or an antigen-binding fragment thereof as described in any one of claims 1-7.

9. An expression vector comprising the nucleic acid molecule of claim 8.

10. A host cell comprising the nucleic acid molecule of claim 8 and / or the expression vector of claim 9; wherein the host cell is selected from prokaryotic cells and eukaryotic cells.

11. The host cell according to claim 10, characterized in that, The prokaryotic cells include bacterial cells.

12. The host cell according to claim 11, characterized in that, The bacterial cells include Escherichia coli and Streptomyces.

13. The host cell according to claim 10, characterized in that, The eukaryotic cells include yeast cells, mammalian cells, and insect cells.

14. The host cell according to claim 13, characterized in that, The mammals are selected from humans, monkeys, mice, rats, hamsters, goats, sheep, cattle, pigs, dogs, or cats.

15. The host cell according to claim 13, characterized in that, The mammalian cells include CHO cells, 293 cells, Vero cells, BHK cells, or PER.C6 cells.

16. A chimeric antigen receptor comprising an extracellular antigen-binding domain, a transmembrane domain, and an intracellular signal transduction domain, wherein the extracellular antigen-binding domain comprises the antibody or an antigen-binding fragment thereof as described in any one of claims 1-7.

17. A modified immune cell comprising the chimeric antigen receptor of claim 16.

18. A detection kit comprising the antibody or antigen-binding fragment thereof as described in any one of claims 1-7.

19. The use of the antibody or antigen-binding fragment thereof of any one of claims 1-7, the nucleic acid molecule of claim 8, the expression vector of claim 9, the host cell of any one of claims 10-15, the chimeric antigen receptor of claim 16, the modified immune cell of claim 17, and the detection kit of claim 18 in the preparation of products for the diagnosis, prevention, and / or treatment of human papillomavirus infection, wherein the human papillomavirus is selected from HPV16, HPV18, HPV31, HPV33, HPV52, or HPV58.

20. A composition comprising (A1) and (A2), and a pharmaceutically acceptable carrier, in, (A1) is an anti-human papillomavirus antibody or its antigen-binding fragment, (A1) comprising: LCDR1, LCDR2, and LCDR3 contained in the light chain variable region as shown in SEQ ID NO: 20; and HCDR1, HCDR2, and HCDR3 contained in the heavy chain variable region as shown in SEQ ID NO: 23; (A2) is an anti-human papillomavirus antibody or its antigen-binding fragment, (A2) comprising: LCDR1, LCDR2, and LCDR3 contained in the light chain variable region as shown in SEQ ID NO:19; and HCDR1, HCDR2, and HCDR3 contained in the heavy chain variable region as shown in SEQ ID NO:22; LCDR1, LCDR2, LCDR3, HCDR1, HCDR2, and HCDR3 are defined based on the same Kabat, AbM, Chothia, Contact, or IMGT numbering system.

21. The use of the composition of claim 20 in the preparation of products for the diagnosis, prevention and / or treatment of human papillomavirus infection, wherein the human papillomavirus is selected from HPV16, HPV18, HPV31, HPV33, HPV45, HPV52 or HPV58.

22. A composition comprising (A1) and (A3), and a pharmaceutically acceptable carrier, in, (A1) is an anti-human papillomavirus antibody or its antigen-binding fragment, (A1) comprising: LCDR1, LCDR2, and LCDR3 contained in the light chain variable region as shown in SEQ ID NO: 20; and HCDR1, HCDR2, and HCDR3 contained in the heavy chain variable region as shown in SEQ ID NO: 23; (A3) is an anti-human papillomavirus antibody or its antigen-binding fragment, (A3) comprising: LCDR1, LCDR2, and LCDR3 contained in the light chain variable region as shown in SEQ ID NO:21; and HCDR1, HCDR2, and HCDR3 contained in the heavy chain variable region as shown in SEQ ID NO:24; LCDR1, LCDR2, LCDR3, HCDR1, HCDR2, and HCDR3 are defined based on the same Kabat, AbM, Chothia, Contact, or IMGT numbering system.

23. The use of the composition of claim 22 in the preparation of products for the diagnosis, prevention and / or treatment of human papillomavirus infection, wherein the human papillomavirus is selected from HPV6, HPV11, HPV16, HPV18, HPV31, HPV33, HPV52 or HPV58.

24. A composition comprising (A1), (A2) and (A3), and a pharmaceutically acceptable carrier. in, (A1) is an anti-human papillomavirus antibody or its antigen-binding fragment, (A1) comprising: LCDR1, LCDR2, and LCDR3 contained in the light chain variable region as shown in SEQ ID NO: 20; and HCDR1, HCDR2, and HCDR3 contained in the heavy chain variable region as shown in SEQ ID NO: 23; (A2) is an anti-human papillomavirus antibody or its antigen-binding fragment, (A2) comprising: LCDR1, LCDR2, and LCDR3 contained in the light chain variable region as shown in SEQ ID NO:19; and HCDR1, HCDR2, and HCDR3 contained in the heavy chain variable region as shown in SEQ ID NO:22; (A3) is an anti-human papillomavirus antibody or its antigen-binding fragment, (A3) comprising: LCDR1, LCDR2, and LCDR3 contained in the light chain variable region as shown in SEQ ID NO:21; and HCDR1, HCDR2, and HCDR3 contained in the heavy chain variable region as shown in SEQ ID NO:24; LCDR1, LCDR2, LCDR3, HCDR1, HCDR2, and HCDR3 are defined based on the same Kabat, AbM, Chothia, Contact, or IMGT numbering system.

25. The use of the composition of claim 24 in the preparation of products for the diagnosis, prevention and / or treatment of human papillomavirus infection, wherein the human papillomavirus is selected from HPV6, HPV11, HPV16, HPV18, HPV31, HPV33, HPV45, HPV52 or HPV58.

26. A method for detecting human papillomavirus, the method comprising the step of using an antibody or antigen-binding fragment thereof as described in any one of claims 1-7 for detection, the method being a non-diagnostic method, wherein the human papillomavirus is selected from HPV16, HPV18, HPV31, HPV33, HPV52 or HPV58.

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