Anti-human cytomegalovirus antibodies and uses thereof

Abstract

The present disclosure relates to an anti-human cytomegalovirus antibody and its use. The anti-human cytomegalovirus antibody can bind to CMV trimer and / or pentamer complexes, and in particular, is an antibody that completely targets the pentamer-cell receptor binding region, which can completely block the binding of the virus to cells and has important clinical significance.

Description

[0001] This application is a divisional application. The application date of the original application is October 27, 2022, the application number is 2022113262365, and the name of the invention is “Anti-human cytomegalovirus antibodies and their uses”. Technical Field

[0002] The present disclosure relates to antibodies or antigen-binding fragments thereof that are specific for and bind with high affinity to human cytomegalovirus, as well as methods for producing such antibodies. The invented antibodies also have high potency in neutralizing infection. The invention also relates to the epitopes to which the antibodies bind, and the use of the antibodies for diagnosis, prevention, and treatment of infected individuals. Background Art

[0003] Human cytomegalovirus (HCMV) is a ubiquitous pathogen belonging to the β-herpesvirus subfamily of the Herpesviridae family, also known as human herpesvirus 5 (HHV5). It usually presents asymptomatic infection in immunocompetent individuals and causes serious complications only in immunocompromised individuals. The incidence of CMV infection in adults varies widely, ranging from 36% to 77% of the population in developed countries to generally above 90% in developing countries (1). In immunocompromised individuals, reinfection or reactivation of CMV can cause serious complications and even be life-threatening (2,3). In addition, congenital HCMV infection is the main cause of visual / hearing impairment or intellectual disability in 0.7% of newborns (4,5).

[0004] Membrane fusion of human herpesviruses is a complex process that requires highly conserved envelope glycoproteins gB and gH / gL as well as multiple glycoprotein complexes to mediate viral attachment, fusion, or endocytosis to host cells (6,7). Currently, the function of HCMV glycoprotein gB is considered to be the primary fusion complex mediating membrane fusion between the virus and infected cells, which is triggered after the binding of the trimeric or pentameric complex to the receptor (8-10). Two key gH / gL-containing complexes encoded by HCMV are essential for cell entry. The HCMV trimeric complex is formed by gH / gL / gO, in which gL-Cys144 is linked to gO-Cys351 by a disulfide bond. The HCMV pentameric complex contains five different subunits gH / gL / UL128 / UL130 / UL131A, and gL-Cys144 is bound to UL128-Cys162 by a disulfide bond. Because gO-Cys351 in the trimer and UL128-Cys162 in the pentamer share the same binding site with gL-Cys144, the formation of pentamers and trimers is mutually exclusive (11). In addition, the ratio of pentamers to trimers on the viral membrane is regulated by UL148 and US16 (12,13), and UL116 has recently been shown to act as a gH chaperone during the assembly and maturation of the gH complex in infected cells and also influences the formation of the gH complex (14,15). Genetic studies have clearly demonstrated that pentamers are required for entry into epithelial and endothelial cells, but not for entry into fibroblasts. More importantly, UL128 / UL130 / UL131A in the pentamer can maintain the tropism of HCMV for non-fibroblast cells (16,17). In contrast, trimers are thought to be sufficient to mediate CMV entry into fibroblasts. Several studies have shown that gO is essential for maintaining cell-free viral infectivity and that gO-deficient viruses infect neither fibroblasts nor epithelial and endothelial cells (18-20).

[0005] HCMV was first isolated in 1956 (21). Although experimental vaccines and therapeutic monoclonal antibodies (mAbs) have entered clinical trials, there are currently no approved preventive vaccines or therapeutic mAbs for clinical use. To date, CMV gB protein as an immunogen and MF59 as an adjuvant (gB / MF59) are the best performing CMV vaccine candidates in clinical trials, with protection rates of 43% to 50% in phase II clinical trials in solid organ transplant recipients (SOT) (22). Chemotherapeutic drugs such as GCV, Letermovir, MBV, and CMV hyperimmune globulin (CMVIG) have shown efficacy against HCMV (23-25). CMVIG has shown good efficacy after SOT or allogeneic hematopoietic cell transplantation (26-28). After initial infection in the first trimester of pregnancy, CMVIG administered every two weeks effectively prevents mother-to-child transmission of HCMV (29). However, current treatments are severely limited by toxicity or resistance to chemotherapeutic drugs and the shortcomings of blood-derived CMVIG products (30,31). MAb-based therapy has the advantage of overcoming the limitations of currently available drugs and could be an effective clinical alternative. Pentamer-specific mAbs can block HCMV infection of trophoblast progenitor cells (32), and increased levels of antibodies targeting pUL128L within 30 days of HCMV infection in pregnant women are associated with a reduced risk of viral transmission to the fetus (33). In addition, mAbs with the ability to neutralize CMV in epithelial cells can protect solid organ transplant recipients (34). At the same time, gH-specific mAbs have shown broad-spectrum properties in inhibiting viral infection and transmission (35). A recent phase II clinical trial showed that vaccination with Genentech's RG7667 (including pentamer-specific and anti-GH antibodies) in high-risk renal transplant recipients delayed the emergence of CMV virus and resulted in less CMV disease compared with placebo (36). Therefore, antibody cocktail therapy consisting of pentamer-specific and gH-specific mAbs may lead to a breakthrough in CMV immunotherapy. However, there are still no approved preventive vaccines or therapeutic monoclonal antibodies (mAbs) for clinical prevention and treatment of HCMV infection.

[0006] HCMV pentamers primarily mediate viral infection of non-fibroblastic cells, including epithelial cells, endothelial cells, and immune cells. Neutralizing antibodies targeting pentamers are more effective in inhibiting viral infection of non-fibroblastic cells than neutralizing antibodies targeting gB protein. Therefore, in the future, neutralizing antibodies targeting pentamers could be combined with neutralizing antibodies targeting gB protein to effectively block viral infection of multiple cell types, potentially offering greater clinical value.

[0007] MSL-109 is a monoclonal antibody targeting the HCMV gH / gL complex that failed in Phase II clinical trials. This antibody cannot bind to free virus and cannot alleviate CMV viremia clinically (37). The binding epitope of antibody 8I21 on the pentamer only partially overlaps with the region where the pentamer binds to cell-associated receptors (43). 8I21 cannot completely block the binding of the virus to the cell receptor, and there is a possibility of viral breakthrough infection. Therefore, developing an antibody that completely targets the pentamer-cell receptor binding region and completely blocks the binding of the virus to cells is of great clinical significance. Summary of the Invention

[0008] The present disclosure identifies and characterizes a group of 8 HCMV pentamer-reactive antibodies, which can be divided into two groups based on their reactivity to CMV trimers and pentamers. The antibodies PC0004, PC0010, PC0012, PC0014, PC0035 and PC0037 in the first group bind to both trimers and pentamers. Experiments have shown that 3 of the 6 antibodies in the first group (PC0012, PC0014 and PC0035) can neutralize HCMV and recognize a highly conserved domain on the gH / gL protein in trimers and pentamers. These antibodies neutralize HCMV not by blocking the binding of CMV to host cells, but by inhibiting the post-adsorption process of the virus into host cells. The antibodies PC0031 and PC0034 in the second group only bind to pentamers. The inventors found that one of the two pentamer-specific antibodies (PC0034) neutralizes HCMV by blocking the adsorption of the virus to host cells. Further analysis showed that among the 214 CMV genome sequences currently collected in the NCBI database, the antigenic site targeted by antibody PC0034 is 100% conserved in the UL128 and UL131A proteins.

[0009] The present disclosure reveals the key residues on the antigenic epitopes targeted by these two groups of potent neutralizing antibodies and provides important information about neutralizing epitopes on trimers or pentamers for the design and development of trimers and / or pentamers as CMV vaccines. The potent neutralizing mAbs provided by the present disclosure can serve as attractive candidate antibodies for the development of "cocktail" antibody therapies for the prevention and treatment of HCMV infection.

[0010] In one aspect, the present disclosure provides an anti-human cytomegalovirus antibody or antigen-binding fragment thereof, comprising a heavy chain and a light chain CDR combination selected from the group consisting of:

[0011] (1) heavy chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs. 3-5, respectively, and light chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs. 8-10, respectively;

[0012] (2) heavy chain CDR1, CDR2, and CDR3 sequences comprising SEQ ID NOs. 13-15, respectively, and light chain CDR1, CDR2, and CDR3 sequences comprising SEQ ID NOs. 18-20, respectively;

[0013] (3) heavy chain CDR1, CDR2, and CDR3 sequences comprising SEQ ID NOs. 23-25, respectively, and light chain CDR1, CDR2, and CDR3 sequences comprising SEQ ID NOs. 28-30, respectively;

[0014] (4) heavy chain CDR1, CDR2, and CDR3 sequences comprising SEQ ID NOs. 33-35, respectively, and light chain CDR1, CDR2, and CDR3 sequences comprising SEQ ID NOs. 38-40, respectively;

[0015] (5) heavy chain CDR1, CDR2, and CDR3 sequences comprising SEQ ID NOs. 43-45, respectively, and light chain CDR1, CDR2, and CDR3 sequences comprising SEQ ID NOs. 48-50, respectively;

[0016] (6) heavy chain CDR1, CDR2, and CDR3 sequences comprising SEQ ID NOs. 53-55, respectively, and light chain CDR1, CDR2, and CDR3 sequences comprising SEQ ID NOs. 58-60, respectively;

[0017] (7) heavy chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs. 63-65, respectively, and light chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs. 68-70, respectively; and

[0018] (8) The heavy chain CDR1, CDR2 and CDR3 sequences comprised of SEQ ID NOs. 73-75, respectively, and the light chain CDR1, CDR2 and CDR3 sequences comprised of SEQ ID NOs. 78-80, respectively.

[0019] In another aspect, the present disclosure provides nucleic acids encoding the aforementioned antibodies, or antigen-binding portions thereof.

[0020] In another aspect, the present disclosure provides a vector comprising the aforementioned nucleic acid.

[0021] In another aspect, the present disclosure provides a host cell comprising the aforementioned nucleic acid or vector.

[0022] In another aspect, the present disclosure provides a pharmaceutical composition comprising the aforementioned antibody or antigen-binding portion thereof, nucleic acid, vector and / or cell.

[0023] In another aspect, the present disclosure provides an immunoconjugate comprising the aforementioned antibody or antigen-binding fragment thereof and a label.

[0024] In another aspect, the present disclosure provides a method for producing the aforementioned antibody or antigen-binding fragment thereof, the method comprising culturing a host cell containing the aforementioned nucleic acid or expression vector encoding the aforementioned antibody.

[0025] In another aspect, the present disclosure provides use of the aforementioned antibodies or antigen-binding portions thereof, nucleic acids, vectors, host cells, or immunoconjugates for preparing pharmaceutical compositions or kits for detecting, treating, preventing and / or alleviating CMV infection or CMV-related diseases.

[0026] In another aspect, the present disclosure provides a method for preventing or treating HCMV infection or HCMV-related disease in a human subject, comprising administering to a subject in need of treatment an effective amount of the aforementioned antibody or antigen-binding portion thereof, nucleic acid, vector, host cell, or immunoconjugate.

[0027] In another aspect, the present disclosure provides a method for improving, enhancing or stimulating resistance in a human subject infected with HCMV, comprising administering to a subject in need thereof an effective amount of the aforementioned antibody or antigen-binding portion thereof, nucleic acid, vector, host cell, or immunoconjugate.

[0028] In another aspect, the present disclosure provides a method of neutralizing HCMV in an individual or sample, comprising contacting the aforementioned antibody or antigen-binding fragment thereof with the individual or sample, and testing the ability of the aforementioned antibody or antigen-binding fragment thereof to bind to neutralize HCMV. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate embodiments consistent with the specification and, together with the description, serve to explain the principles of the specification.

[0030] Figure 1 The figures show the detection results of CMV trimer complex and pentameric complex by high performance liquid chromatography and polyacrylamide gel electrophoresis (SDS-PAGE), wherein A. high performance liquid chromatography detection result; B. polyacrylamide gel electrophoresis detection result.

[0031] Figure 2 Shown are the results of ELISA detection of antibodies that specifically bind to the pentameric complex.

[0032] Figure 3 Binding results are shown for antibodies that bind to CMV trimer and pentameric complexes.

[0033] Figure 4The neutralizing activity of antibodies that specifically bind to CMV pentamers is shown, wherein: A. the results of the antibody neutralizing the HCMV virus strain Towne in human embryonic lung fibroblasts (MRC-5); B. the results of the antibody neutralizing the HCMV virus strain BE13 / 2012 in human embryonic lung fibroblasts (MRC-5); C. the neutralization of HCMV virus strains VR1814, NR and AD169 FIX in ARPE-19 epithelial cells.

[0034] Figure 5 Binding results for antibodies specifically binding to CMV trimer and pentamer complexes are shown, with A. neutralizing antibody binding to pentamer; B. neutralizing antibody binding to trimer. In Panel A, the highest response curve corresponds to a pentamer concentration of 6 μg / mL or 4 μg / mL, the lowest response curve corresponds to a pentamer concentration of 0.375 μg / mL or 0.25 μg / mL, and the intermediate response curve corresponds to a two-fold serial dilution of the pentamer concentration. In Panel B, the highest response curve corresponds to a trimer concentration of 4 μg / mL, the lowest response curve corresponds to a trimer concentration of 0.25 μg / mL, and the intermediate response curve corresponds to a two-fold serial dilution of the trimer concentration.

[0035] Figure 6 Shown are the results of cross-competitive blocking of binding of neutralizing antibodies to CMV pentamers.

[0036] Figure 7 The binding results of neutralizing antibodies to pentamer mutants are shown, including: A. the binding results of neutralizing antibodies to pentamer mutants; B. the effect of neutralizing antibodies on CMV virus adhesion to host cells; C. the effect of neutralizing antibodies on CMV-infected cells; D. the effect of neutralizing antibodies on the spread of CMV.

[0037] Figure 8 Figure 2 shows antibody PC0034 blocking HCMV pentamer binding to epithelial and endothelial cells. A. Binding results of antibodies PC0034 and 9I6 to HCMV pentamers. The highest response curve in the figure corresponds to a pentamer concentration of 4 μg / mL, the lowest response curve corresponds to a pentamer concentration of 0.25 μg / mL, and the intermediate response curve corresponds to two-fold serial dilutions of the pentamer concentration. B. SPR analysis results of antibodies PC0034 and 9I6; C. Binding of HCMV pentamers to epithelial cells (APRE-19) and endothelial cells (HUVEC); D. Antibody PC0034 inhibits pentamer binding to epithelial and endothelial cells.

[0038] Figure 9The binding results of antibodies to HCMV pentamer mutants are shown, including: A. Binding results of antibody PC0034 to pentamer mutant UL131A_E23A. The highest response curve in the figure corresponds to a pentamer mutant concentration of 80 μg / mL, the lowest response curve corresponds to a pentamer mutant concentration of 5 μg / mL, and the intermediate response curve corresponds to a two-fold dilution of the pentamer mutant concentration. B. Binding results of antibody PC0034 to pentamer mutant UL131A_K27A. The highest response curve in the figure corresponds to a pentamer mutant concentration of 40 μg / mL, the lowest response curve corresponds to a pentamer mutant concentration of 2.5 μg / mL, and the intermediate response curve corresponds to a two-fold dilution of the pentamer mutant concentration. C. Binding results of antibody PC0034 to pentamer mutant UL128_K47A. The highest response curve in the figure corresponds to a 60 μg / mL concentration of the pentamer mutant, the lowest response curve corresponds to a 3.75 μg / mL concentration of the pentamer mutant, and the intermediate response curve corresponds to a two-fold serial dilution of the pentamer mutant. D. Binding results of antibody PC0034 to the pentamer mutant UL128_T94A. The highest response curve in the figure corresponds to a 40 μg / mL concentration of the pentamer mutant, the lowest response curve corresponds to a 2.5 μg / mL concentration of the pentamer mutant, and the intermediate response curve corresponds to a two-fold serial dilution of the pentamer mutant. E. Binding results of antibody PC0034 to the pentamer. The highest response curve in the figure corresponds to a 20 μg / mL concentration of the pentamer, the lowest response curve corresponds to a 1.25 μg / mL concentration of the pentamer, and the intermediate response curve corresponds to a two-fold serial dilution of the pentamer. F. Binding results of the pentamer mutant to epithelial cells. The highest response curve in the figure corresponds to a pentamer concentration of 20 μg / mL, the lowest response curve corresponds to a pentamer concentration of 1.25 μg / mL, and the intermediate response curve corresponds to a two-fold serial dilution of the pentamer concentration. G. Binding results of pentamer mutants to epithelial cells; H and I. Distribution of K27 on subunit UL131A and T94 and K47 on subunit UL128 within the pentamer complex. DETAILED DESCRIPTION

[0039] I. Definition

[0040] In this disclosure, unless otherwise indicated, scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. Furthermore, terms and laboratory procedures related to protein and nucleic acid chemistry, molecular biology, cell and tissue culture, microbiology, and immunology used herein are those widely used in the respective fields and are common procedures. To facilitate a better understanding of this disclosure, definitions and explanations of relevant terms are provided below.

[0041] To interpret this specification, the following definitions will apply, and wherever appropriate, terms used in the singular may also include the plural, and vice versa. It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.

[0042] The term "about" when used in conjunction with a numerical value is meant to encompass the numerical value within a range having a lower limit that is 5% less than the specified numerical value and an upper limit that is 5% greater than the specified numerical value.

[0043] As used herein, the term "and / or" means any one of the alternatives or two or more of the alternatives.

[0044] As used herein, the terms "comprising" or "including" are intended to include the stated elements, integers, or steps, but do not exclude any other elements, integers, or steps. In this document, when the terms "comprising" or "including" are used, unless otherwise indicated, the context of consisting of the stated elements, integers, or steps is also encompassed. For example, when reference is made to an antibody variable region "comprising" a specific sequence, it is intended to encompass an antibody variable region consisting of that specific sequence.

[0045] "Human cytomegalovirus" (HCMV) is a DNA double-helical virus of the genus Cytomegalovirus in the subfamily Herpesvirinae, also known as human herpersvirus 5 (HHV-5). As used herein, "human cytomegalovirus," "HCMV," "human herpesvirus 5," and "HHV-5" are all interchangeable.

[0046] The term "antibody" is used in the broadest sense herein and encompasses a variety of antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies) and antibody fragments, as long as they show the desired antigen-binding activity. A complete antibody will typically comprise at least two full-length heavy chains and two full-length light chains, but may comprise fewer chains in certain cases, such as antibodies naturally occurring in camels that may comprise only heavy chains. The antibody may be a humanized or human antibody and a single domain antibody, such as VH, VHH or VL. Examples of antibody fragments include but are not limited to Fab, Fab', F(ab')2, single-chain Fv (scFv), Fv, dsFv, diabodies, Fd and Fd' fragments and other fragments, including modified fragments (e.g., Methods in Molecular Biology, Vol 207: Recombinant Antibodies for Cancer Therapy Methods and Protocols (2003); Chapter 1; p3-25, Kipriyanov). Such fragments may comprise multiple chains linked together, for example, by disulfide bonds and / or by peptide linkers.Antibody fragments generally contain at least or about 50 amino acids, and typically at least or about 200 amino acids.

[0047] The term "complementarity determining region" or "CDR region" or "CDR" or "hypervariable region" refers to the amino acid region in the variable region of an antibody that is primarily responsible for binding to the antigen epitope. The CDRs of the heavy and light chains are usually referred to as CDR1, CDR2, and CDR3, and are numbered sequentially starting from the N-terminus.

[0048] A variety of methods are known in the art for determining CDR sequences in a given VH or VL amino acid sequence: Kabat complementarity determining regions (CDRs) are determined based on sequence variability and are the most commonly used (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Edition, Public Health Service, National Institutes of Health, Bethesda, Md. (1991)), while Chothia refers to the position of structural loops (Chothia et al., (1987) J. Mol. Biol. 196: 901-917; Chothia et al. (1989) Nature 342: 877-883). AbM CDRs are a compromise between Kabat CDRs and Chothia structural loops and are used by Oxford Molecular's AbM antibody modeling software. "Contact" CDRs are based on analysis of available complex crystal structures. The residues for each of these CDRs, according to different CDR determination schemes, are shown below.

[0049] Table 1 CDR residue scheme

[0050]

[0051]

[0052] A CDR can also be identified based on having the same Kabat numbering position as a reference CDR sequence (eg, any of the exemplary CDRs of the invention).

[0053] Unless otherwise indicated, in the present invention, the term "CDR" or "CDR sequence" encompasses CDR sequences determined in any of the above-mentioned ways.

[0054] Unless otherwise indicated, in the present invention, when referring to residue positions in the variable region of an antibody (including heavy chain variable region residues and light chain variable region residues), the numbering refers to the position according to the Kabat numbering system (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)).

[0055] In one embodiment, the CDRs of an antibody of the invention have boundaries defined by the IMGT rules, for example, using the IMGT database.

[0056] It should be noted that the boundaries of the CDRs of the variable regions of the same antibody obtained based on different assignment systems may be different. That is, the CDR sequences of the variable regions of the same antibody defined under different assignment systems may be different. Therefore, when referring to antibodies defined by specific CDR sequences defined in the present invention, the scope of the antibodies also covers antibodies whose variable region sequences contain the specific CDR sequences, but whose claimed CDR boundaries are different from the specific CDR boundaries defined in the present invention due to the application of different schemes (e.g., different assignment system rules or combinations).

[0057] The term "variant" associated with an antibody refers herein to an antibody comprising an antibody having an amino acid change in a target antibody region (e.g., a heavy chain variable region or a light chain variable region or a heavy chain CDR region or a light chain CDR region) that has been replaced, deleted, and / or inserted by at least one amino acid residue (e.g., 1-30, or 1-20 or 1-10, e.g., 1 or 2 or 3 or 4 or 5 amino acid residues), or by chemically derivatizing one or more amino acid residues, wherein the variant substantially retains the biological properties of the antibody molecule before the change. On the one hand, the present disclosure encompasses variants of any antibody mentioned herein. In one embodiment, the antibody variant retains at least 60%, 70%, 80%, 90%, or 100% of the biological activity (e.g., antigen binding ability) of the antibody before the change. It will be appreciated that the heavy chain variable region or light chain variable region, or each CDR region of an antibody can be changed individually or in combination. In some embodiments, the amino acid changes in one or more or all three heavy chain CDRs are no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. Preferably, the above-mentioned amino acid changes are amino acid substitutions, preferably conservative substitutions. In some embodiments, the antibody variants have at least greater than 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or higher sequence identity with the parent antibody over the region of the antibody sequence of interest.

[0058] The term "conservative substitution" refers to the substitution of one amino acid by another amino acid within the same class, for example, an acidic amino acid by another acidic amino acid, a basic amino acid by another basic amino acid, or a neutral amino acid by another neutral amino acid. Exemplary substitutions are shown in Table 2 below:

[0059] Table 2 Conservative amino acid substitutions

[0060]

[0061]

[0062] As used herein, the term "neutralize" refers to neutralizing the ability of a pathogen to initiate and / or maintain an infection in a host.

[0063] As used herein, the term "epitope" refers to a portion of an antigen (e.g., the gB glycoprotein of HCMV) that specifically interacts with an antibody molecule. An epitope within a protein antigen can be formed from continuous amino acids (typically linear epitopes) or discontinuous amino acids (typically conformational epitopes) juxtaposed by the tertiary folding of the protein. Epitopes formed by continuous amino acids are typically (but not always) exposed to denaturing solvents, while epitopes formed by tertiary folding are typically lost when treated with denaturing solvents.

[0064] An "antibody that binds to the same or overlapping epitope as a reference antibody" is an antibody that blocks 50%, 60%, 70%, 80%, 90% or 95% or more of the binding of the reference antibody to its antigen in a competition assay, whereas conversely, the reference antibody blocks 50%, 60%, 70%, 80%, 90% or 95% or more of the binding of the antibody to its antigen in a competition assay.

[0065] An antibody that competes with a reference antibody for binding to its antigen is one that blocks 50%, 60%, 70%, 80%, 90% or more of the binding of the reference antibody to its antigen in a competition assay. Conversely, a reference antibody blocks 50%, 60%, 70%, 80%, 90% or more of the binding of the antibody to its antigen in a competition assay. Numerous types of competition assays can be used to determine whether one antibody competes with another, such as ELISA, SPR, solid phase direct or indirect radioimmunoassay (RIA), solid phase direct or indirect enzyme immunoassay (EIA) (see, e.g., Stahli et al., 1983, Methods in Enzymology 9: 242-253).

[0066] An antibody that inhibits (e.g., competitively inhibits) the binding of a reference antibody to its antigen is an antibody that inhibits the binding of the reference antibody to its antigen by 50%, 60%, 70%, 80%, 90%, or 95% or more. Conversely, the reference antibody inhibits the binding of the antibody to its antigen by 50%, 60%, 70%, 80%, 90%, or 95% or more. The binding of an antibody to its antigen can be measured by affinity (e.g., equilibrium dissociation constant). Methods for determining affinity are known in the art.

[0067] An antibody that exhibits the same or similar binding affinity and / or specificity as a reference antibody is an antibody that has at least 50%, 60%, 70%, 80%, 90% or more than 95% of the binding affinity and / or specificity of the reference antibody. This can be determined by any method known in the art for determining binding affinity and / or specificity.

[0068] An "IgG-type antibody" refers to an antibody whose heavy chain constant region belongs to the IgG type. All antibodies of the same type have the same heavy chain constant region, but different antibodies of different types have different heavy chain constant regions. For example, an IgG1-type antibody refers to an antibody whose heavy chain constant region Ig domain is the IgG3 domain of IgG1.

[0069] A "human" antibody (HuMAb) is an antibody having variable regions in which both the framework and CDR regions are derived from human germline immunoglobulin sequences. In addition, if the antibody contains a constant region, the constant region is also derived from human germline immunoglobulin sequences.

[0070] A "humanized" antibody refers to an antibody in which some, most or all of the amino acids outside the CDR domains of a non-human antibody (e.g., a mouse antibody) are replaced by corresponding amino acids derived from human immunoglobulins. In one embodiment of a humanized form of an antibody, some, most or all of the amino acids outside the CDR domains have been replaced by amino acids from human immunoglobulins, while some, most or all of the amino acids within one or more CDR regions have not changed. Small additions, deletions, insertions, substitutions or modifications of amino acids are permitted as long as they do not eliminate the ability of the antibody to bind to a specific antigen. A "humanized" antibody retains an antigenic specificity similar to that of the original antibody.

[0071] As used herein, a "chimeric antibody" refers to an antibody in which the variable region is derived from one species and the constant region is derived from another species, such as an antibody in which the variable region is derived from a mouse antibody and the constant region is derived from a human antibody.

[0072] As used herein, "antibody fragment" refers to a molecule different from an intact antibody, which comprises a portion of an intact antibody and binds to the antigen to which the intact antibody binds. As used herein, the term "antigen-binding fragment" refers to one or more fragments of an antibody that retains the ability to specifically bind to an antigen (e.g., gB glycoprotein of human HCMV). Examples of antibody fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab')2, diabodies, linear antibodies, single-chain antibodies (e.g., scFv), single-domain antibodies, bivalent or bispecific antibodies or fragments thereof, camelid antibodies, and bispecific antibodies or multispecific antibodies formed from antibody fragments.

[0073] As used herein, "multispecific" refers to an antibody that specifically binds to at least two different antigens or two different epitopes within an antigen, such as three, four, or five different antigens or epitopes.

[0074] As used herein, "bispecific" refers to an antibody that specifically binds to two different antigens or two different epitopes within the same antigen. A bispecific antibody may have cross-reactivity to other related antigens, or may bind to an epitope shared between two or more different antigens.

[0075] An "immunoconjugate" is an antibody conjugated to one or more additional substances, including but not limited to a label.

[0076] The term "label" as used herein refers to a compound or composition that is directly or indirectly conjugated or fused to a reagent (such as a polynucleotide probe or antibody) and promotes the detection of the reagent to which it is conjugated or fused. The label itself can be detectable (e.g., radioisotope labeling or fluorescent labeling) or can catalyze the chemical alteration of a detectable substrate compound or composition in the case of an enzymatic labeling. The term is intended to encompass direct labeling of a probe or antibody by coupling (i.e., physically connecting) a detectable substance to the probe or antibody and indirect labeling of a probe or antibody by reacting with another reagent of direct labeling. Examples of indirect labeling include detection of a primary antibody using a fluorescently labeled secondary antibody and end-labeling of a DNA probe with biotin so that it can be detected with fluorescently labeled streptavidin.

[0077] The term "isolated" antibody refers to an antibody that has been separated from the components of its natural environment. In some embodiments, the antibody is purified to greater than 95% or 99% purity, as determined by, for example, electrophoresis (e.g., SDS-PAGE, isoelectric focusing (IEF), capillary electrophoresis) or chromatography (e.g., ion exchange or reversed-phase HPLC). For a review of methods for assessing antibody purity, see, e.g., Flatman et al., J. Chromatogr. B 848:79-87 (2007).

[0078] The term "isolated" nucleic acid refers to a nucleic acid molecule that has been separated from a component of its natural environment. An isolated nucleic acid includes a nucleic acid molecule contained in cells that normally contain the nucleic acid molecule, but the nucleic acid molecule is present extrachromosomally or at a chromosomal location that is different from its natural chromosomal location.

[0079] The term "affinity" or "binding affinity" refers to the intrinsic binding affinity that reflects the interaction between members of a binding pair (e.g., an antibody and an antigen). The affinity of a molecule X for its partner Y can usually be expressed in terms of the equilibrium dissociation constant (KD). The equilibrium dissociation constant is the ratio of the dissociation rate constant and the association rate constant (kdis and kon, respectively). The smaller the KD, the smaller the dissociation, which represents the stronger the affinity between the antibody and the antigen. Affinity can be measured by common methods known in the art, for example, using surface plasmon resonance (SPR) in a BIACORE instrument. Typically, antibodies (e.g., the neutralizing antibody TRN1021 of the present disclosure) are measured at a KD of no more than 1×10 -5 M, for example, is less than about 1×10 -6 M, 1×10 -7 M, 1×10 -8 M, 1×10 -9 M or 1×10 -10 The molecule dissociates from the antigen with an equilibrium dissociation constant (KD) of M or less.

[0080] The term "immunoconjugate" refers to an antibody conjugated to one or more heterologous molecules, including but not limited to a carrier. The term "pharmaceutical composition" refers to a preparation that is in a form that permits the biological activity of the active ingredient contained therein to be effective and does not contain additional ingredients that are unacceptably toxic to a subject to which the preparation is administered.

[0081] The term "pharmaceutically acceptable carrier" refers to one or more non-toxic materials that are administered with a therapeutic agent and do not interfere with the biological activity of the active ingredient, including but not limited to buffers, preservatives, compatible carriers, diluents, adjuvants (e.g., Freund's adjuvant (complete and incomplete)), excipients, vehicles, and optionally other additives or encapsulating materials. Pharmaceutical carriers suitable for use in the present disclosure can be conventional pharmaceutical formulation excipients; and compositions and formulations suitable for delivering the disclosed neutralizing antibodies.

[0082] The term "pharmaceutical composition" refers to a composition that is in form permitting the biological activity of the active ingredient contained therein to be effective, and that contains no additional ingredients that are unacceptably toxic to a subject to which the composition would be administered.

[0083] The term "effective amount" refers to an amount or dosage of an antibody, fragment, conjugate, or composition of the invention that produces the desired effect in a patient in need of treatment or prevention after administration to the patient in single or multiple doses. The effective amount can be readily determined by the attending physician, who is skilled in the art, by considering a variety of factors, such as the species of the mammal; its size, age, and general health; the specific disease involved; the extent or severity of the disease; the response of the individual patient; the specific antibody administered; the mode of administration; the bioavailability characteristics of the administered formulation; the selected dosing regimen; and the use of any concomitant therapy.

[0084] A "therapeutically effective amount" refers to an amount effective to achieve the desired therapeutic outcome at the desired dosage and for the desired period of time. A therapeutically effective amount of an antibody or antibody fragment, or conjugate or composition thereof, can vary depending on a variety of factors, such as the disease state, age, sex, and weight of the individual, and the ability of the antibody or antibody portion to elicit a desired response in the individual. A therapeutically effective amount is also an amount in which any toxic or deleterious effects of the antibody or antibody fragment, or conjugate or composition thereof, are outweighed by the therapeutically beneficial effects. A "therapeutically effective amount" preferably inhibits a measurable parameter by at least about 20%, more preferably at least about 40%, even more preferably at least about 50%, 60%, or 70%, and still more preferably at least about 80% or 90%, relative to an untreated individual. The ability of a compound to inhibit a measurable parameter can be evaluated in an animal model system that is predictive of efficacy in human autoimmune disease or inflammation.

[0085] A "prophylactically effective amount" refers to an amount effective to achieve the desired preventive result, at the required dosage and for the required period of time. Typically, a prophylactic amount will be less than a therapeutically effective amount because a prophylactic dose is used in an individual prior to or at an earlier stage of disease.

[0086] As used herein, "individual" or "subject" are used interchangeably and include mammals, such as humans.

[0087] As used herein, "treat," ...

[0088] The term "vaccine" or "vaccine composition" refers to a composition comprising at least one immunogenic composition that induces an immune response in an animal.

[0089] The term "subject" or "individual" is a primate (eg, a human and a non-human primate such as a monkey). In certain embodiments, the individual or subject is a human.

[0090] II. Detailed description of specific implementation plan

[0091] In one aspect, the present disclosure provides an anti-human cytomegalovirus antibody or antigen-binding fragment thereof, comprising a heavy chain and a light chain CDR combination selected from the group consisting of:

[0092] (1) heavy chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs. 3-5, respectively, and light chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs. 8-10, respectively;

[0093] (2) heavy chain CDR1, CDR2, and CDR3 sequences comprising SEQ ID NOs. 13-15, respectively, and light chain CDR1, CDR2, and CDR3 sequences comprising SEQ ID NOs. 18-20, respectively;

[0094] (3) heavy chain CDR1, CDR2, and CDR3 sequences comprising SEQ ID NOs. 23-25, respectively, and light chain CDR1, CDR2, and CDR3 sequences comprising SEQ ID NOs. 28-30, respectively;

[0095] (4) heavy chain CDR1, CDR2, and CDR3 sequences comprising SEQ ID NOs. 33-35, respectively, and light chain CDR1, CDR2, and CDR3 sequences comprising SEQ ID NOs. 38-40, respectively;

[0096] (5) heavy chain CDR1, CDR2, and CDR3 sequences comprising SEQ ID NOs. 43-45, respectively, and light chain CDR1, CDR2, and CDR3 sequences comprising SEQ ID NOs. 48-50, respectively;

[0097] (6) heavy chain CDR1, CDR2, and CDR3 sequences comprising SEQ ID NOs. 53-55, respectively, and light chain CDR1, CDR2, and CDR3 sequences comprising SEQ ID NOs. 58-60, respectively;

[0098] (7) heavy chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs. 63-65, respectively, and light chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs. 68-70, respectively; and

[0099] (8) The heavy chain CDR1, CDR2 and CDR3 sequences comprised of SEQ ID NOs. 73-75, respectively, and the light chain CDR1, CDR2 and CDR3 sequences comprised of SEQ ID NOs. 78-80, respectively.

[0100] In some embodiments, the aforementioned antibodies, or antigen-binding portions thereof, comprise a combination of heavy chain variable regions and light chain variable regions selected from the group consisting of:

[0101] (1) the heavy chain variable region comprises or consists of an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO. 1, and the light chain variable region comprises or consists of an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO. 6;

[0102] (2) the heavy chain variable region comprises or consists of an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO. 11, and the light chain variable region comprises or consists of an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO. 16;

[0103] (3) the heavy chain variable region comprises or consists of an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO. 21, and the light chain variable region comprises or consists of an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO. 26;

[0104] (4) the heavy chain variable region comprises or consists of an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO. 31, and the light chain variable region comprises or consists of an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO. 36;

[0105] (5) the heavy chain variable region comprises or consists of an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO. 41, and the light chain variable region comprises or consists of an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO. 46;

[0106] (6) the heavy chain variable region comprises or consists of an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO. 51, and the light chain variable region comprises or consists of an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO. 56;

[0107] (7) the heavy chain variable region comprises or consists of an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO. 61, and the light chain variable region comprises or consists of an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO. 66;

[0108] (8) The heavy chain variable region comprises or consists of an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO. 71, and the light chain variable region comprises or consists of an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO. 76.

[0109] In some embodiments, the aforementioned antibody is an IgG1, IgG2, IgG3, or IgG4 antibody.

[0110] In some embodiments, the aforementioned antibody is an IgG1 antibody.

[0111] In some embodiments, the constant region of the aforementioned heavy chain comprises or consists of an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence shown in SEQ ID NO.81.

[0112] In some embodiments, the constant region of the aforementioned light chain comprises or consists of an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence shown in SEQ ID NO. 89 or 91.

[0113] In some embodiments, the aforementioned nucleic acid molecule comprises an antibody heavy chain variable region nucleic acid sequence selected from SEQ ID NO. 2, 12, 22, 32, 42, 52, 62, 72 or any variant thereof, and an antibody light chain variable region nucleic acid sequence selected from SEQ ID NO. 7, 17, 27, 37, 47, 57, 67, 77 or any variant thereof.

[0114] In another aspect, the present disclosure provides nucleic acids encoding the aforementioned antibodies, or antigen-binding portions thereof.

[0115] In another aspect, the present disclosure provides a vector comprising the aforementioned nucleic acid.

[0116] In another aspect, the present disclosure provides a host cell comprising the aforementioned nucleic acid or vector.

[0117] In another aspect, the present disclosure provides a pharmaceutical composition comprising the aforementioned antibody or antigen-binding portion thereof, nucleic acid, vector and / or cell.

[0118] In another aspect, the present disclosure provides an immunoconjugate comprising the aforementioned antibody or antigen-binding fragment thereof and a label.

[0119] In another aspect, the present disclosure provides a method for producing the aforementioned antibody or antigen-binding fragment thereof, the method comprising culturing a host cell containing the aforementioned nucleic acid or expression vector encoding the aforementioned antibody.

[0120] In another aspect, the present disclosure provides use of the aforementioned antibodies or antigen-binding portions thereof, nucleic acids, vectors, host cells, or immunoconjugates for preparing pharmaceutical compositions or kits for detecting, treating, preventing and / or alleviating CMV infection or CMV-related diseases.

[0121] In some embodiments, the CMV is HCMV.

[0122] In another aspect, the present disclosure provides a method for preventing or treating HCMV infection or HCMV-related disease in a human subject, comprising administering to a subject in need of treatment an effective amount of the aforementioned antibody or antigen-binding portion thereof, nucleic acid, vector, host cell, or immunoconjugate.

[0123] In another aspect, the present disclosure provides a method for improving, enhancing or stimulating resistance in a human subject infected with HCMV, comprising administering to a subject in need thereof an effective amount of the aforementioned antibody or antigen-binding portion thereof, nucleic acid, vector, host cell, or immunoconjugate.

[0124] In some embodiments, the individual is infected with HCMV.

[0125] In another aspect, the present disclosure provides a method of neutralizing HCMV in an individual or sample, comprising contacting the aforementioned antibody or antigen-binding fragment thereof with the individual or sample, and testing the ability of the aforementioned antibody or antigen-binding fragment thereof to bind to neutralize HCMV.

[0126] The aforementioned anti-human cytomegalovirus antibodies can bind to CMV trimer and / or pentamer complexes, especially antibodies that completely target the pentamer and cell receptor binding region, which can completely block the binding of the virus to cells and have important clinical significance.

[0127] For purposes of clarity and conciseness, features are described herein as part of the same or separate embodiments; however, it will be understood that the scope of the present disclosure may include embodiments having a combination of all or some of the described features.

[0128] Example

[0129] Example 1 Expression, purification and identification of CMV recombinant antigen protein

[0130] The UL74 (gO), UL75 (gH), UL115 (gL), UL128, UL130, and UL131A gene sequences of the VR1814 (GU179289.1) strain from NCBI GenBank were selected and codon-optimized. The UL75 (gH) gene was truncated to amino acids 1-715 to remove the transmembrane and intracellular regions for expression of a soluble pentamer protein. The amino acid sequence of the UL74 (gO) antigen is shown in SEQ ID NO.83; the amino acid sequence of the truncated UL75 (gH) antigen protein is shown in SEQ ID NO.84; the amino acid sequence of the UL115 (gL) antigen is shown in SEQ ID NO.85; the amino acid sequence of the UL128 antigen is shown in SEQ ID NO.86; the amino acid sequence of the UL130 antigen is shown in SEQ ID NO.87; and the amino acid sequence of the UL131A antigen is shown in SEQ ID NO.88.

[0131] All antigenic proteins were labeled with six histidine tags at their carboxyl termini and were individually constructed into the pcDNA3.1 eukaryotic expression vector. The coding genes for each subunit of the gH / gL / UL128 / UL130 / UL131A pentamer complex (the mass ratio of each subunit to the plasmid was 1:0.8:0.6:0.6:0.6) and the gH / gL / gO trimer complex (the mass ratio of each subunit to the plasmid was 1:1:1) were mixed and transiently co-transfected into 293i cells. After 5 days of culture, the cell supernatant was collected and purified using Ni-NTA. The expression was detected by high-performance liquid chromatography and polyacrylamide gel electrophoresis (SDS-PAGE) ( Figure 1A and 1B), it was determined that the obtained recombinant protein complex had complete pentamer and trimer structures.

[0132] Example 2 Acquisition of CMV Pentamer and Trimer Reactive Antibodies

[0133] After obtaining the recombinant antigen protein prepared in Example 1, blood samples from healthy volunteers were screened using ELISA to identify samples with high antibody titers against the purified recombinant pentamer complex. PBMCs from blood samples with high antibody titers were flow cytometrically sorted using the dual-color fluorescently labeled pentamer as a probe to obtain single pentamer-specific memory B lymphocytes.

[0134] The immunoglobulin (Ig) variable regions (VHDJH and VLJL) of the heavy and light chain gene segments were amplified from sorted single B cells by RT / nested PCR. The linear expression vector of the antibody was constructed by overlapping PCR, and the recombinantly expressed antibody was obtained by transiently transforming 293T cells (38). A total of 16 antibodies that specifically bound to the pentamer were identified by ELISA binding experiments ( Figure 2 The somatic mutation rates of these antibodies ranged from 2.01-15.38% in the VH gene and 6.47-19.93% in the VL gene, respectively. These 16 antibodies were derived from 11 different Ig gene clones, of which eight (PC0004, PC0031, PC0010, PC0012, PC0014, PC0034, PC0035, and PC0037) represented eight different clones and were selected for production of purified antibodies for further characterization (Table 1). Thus, this example isolated antibodies that specifically bind to HCMV pentamers and trimers from volunteer blood samples.

[0135] Table 1. V(D)J region rearrangements of antibodies specifically binding to HCMV pentamers and trimers

[0136]

[0137] Table 2 Variable regions of anti-human cytomegalovirus antibodies

[0138]

[0139] PC0004

[0140] The VH amino acid sequence is shown in SEQ ID NO.1, its encoding nucleic acid is shown in SEQ ID NO.2, and its HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NOs.3, 4 and 5, respectively.

[0141] VH amino acid sequence:

[0142] EVQLVESGGAMIQPGGSLRLSCAAS GFSFDDYT MYWVRQTPGTGLEWVAL ITWNGVTT RYADSVQGRFTISRDNRKNSSLLQMNSLRPGDSGLYYC ARDIGPLRDSDYYYYGVGV WGLGTTVTVSS

[0143] VH nucleic acid sequence:

[0144] GAGGTTCAGCTGGTGGAGTCTGGGGGAGCCATGATACAGCCGGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCAGTTTTGATGATTATACCATGTATTGGGTCCCGGCAGACTCCGGGGACGGGTCTGGAGTGGGTCGCTCTCATTACTTGGAATGGTGTCACGACAAGATATGCAGACTCT GTGCAGGGCCGATTTACCATCTCCAGAGACAACAGGAAAAACTCTCTGTCTCTGCAAATGAATAGCCTGAGACCTGGGGACAGCGGCTTATATTACTGTGCAAGAGATATCGGCCCCCTACGAGACAGTGACTACTACTACGGTGTGGGCGTCTGGGGCCTAGGGACCACGGTCACCGTCTCCTCA

[0145] The VL amino acid sequence is shown in SEQ ID NO.6, its encoding nucleic acid is shown in SEQ ID NO.7, and its LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NO.8, 9 and 10 respectively. VL amino acid sequence:

[0146] DIVMTQSPLSLPVTPGEPASISCRSS QSLLHINGYNY LHWYLQKPGQSPQLLIY FGS NRASGVSDRFSGSGSGTEFTLKISKVEPEDVGTYYC MQGLQTPLT FGGGTRVEIK

[0147] VL nucleic acid sequence:

[0148] GATATTGTGATGACCCAGTCTCCACTCTCCCTGCCCGTCACCCCTGGAGAGCCGGCCTCCATCTCCTGCAGGTCTAGTCAGAGCCTCCTACATATTAATGGATACAACTATTTGCATTGGTACCTGCAGAAGCCAGGGCAGTCTCCACAGCTCCTGATCTATTTCGGT TCCAATCGGGCCTCCGGGGTCTCTGACAGGTTCAGTGGCAGTGGATCAGGCACAGAGTTCACACTGAAAATTAGTAAAGTTGAGCCTGAGGATGTTGGGACCTATTATTGCATGCAAGGTCTACAAACTCCCCTCACTTTCGGCGGGGGGACGAGGGTGGAGATCAAA

[0149] PC0010

[0150] The VH amino acid sequence is shown in SEQ ID NO.11, its encoding nucleic acid is shown in SEQ ID NO.12, and its HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NO.13, 14 and 15 respectively. VH amino acid sequence:

[0151] QVQLQESGPGLVRPSETLSLMCTVS GASISNTKYY WGWIRQPPGKRLEWVGS LYFSGTT YYNPSLQSRLTMSVDTSKNQFSLNLRSVTAADTAVYYC ARRPFVMSRGVRSDP WGQGILVSVST

[0152] VH nucleic acid sequence:

[0153] CAGGTGCAGCTGCAGGAGTCGGGCCCAGGACTGGTGAGGCCTTCGGAGACCCTGTCCCTCATGTGCACTGTCTCTGGTGCCTCCATCAGCAATACAAAATACTACTGGGGCTGGATCCGCCAGCCCCCAGGGAAGCGACTGGAGTGGGTTGGAAGTCTCTACTTTAGTGGGACCACCTACTACA ACCCGTCCCTCCAGAGTCGACTCACCATGTCCGTAGACACGTCGAAGAACCAGTTCTCCCTCAACCTGAGGTCTGTGACCGCCGCAGACACGGCTGTCTACTATTGTGCGCGACGCCCTTTTGTTATGAGTCGGGGAGTGAGGTCCGACCCCTGGGGCCAGGGAATCCTGGTCTCCGTCTCCACA

[0154] The VL amino acid sequence is shown in SEQ ID NO.16, its encoding nucleic acid is shown in SEQ ID NO.17, and its LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NO.18, 19 and 20 respectively. VL amino acid sequence:

[0155] DIVMTQTPLSLPVTPGEPASISCTSS QSLLQSNGYTY LDWYLQKPGQSPQLLIY LGS NRASGVPDRFSGSGSGTDFTLKISRLEAEDVGVYYC MQALQTPFT FGPGTRVDIK

[0156] VL nucleic acid sequence:

[0157] GATATTGTGATGACCCAGACTCCACTCTCCCTGCCCGTCACCCCTGGAGAGCCGGCCTCCATCTCCTGCACGTCTAGTCAGAGCCTCCTGCAAAGTAATGGATACACCTATTTGGATTGGTACCTGCAGAAGCCAGGGCAGTCTCCACAACTCCTGATCTATTTGGGT TCCAATCGGGCCTCCGGGGTCCCTGACAGGTTCAGTGGCAGTGGATCAGGCACAGATTTTACACTGAAAATCAGTAGACTGGAGGCCGAGGATGTTGGAGTTTATTACTGCATGCAAGCTCTACAAACTCCGTTCACTTTCGGCCCTGGGACCAGAGTGGACATCAAA

[0158] PC0012

[0159] The VH amino acid sequence is shown in SEQ ID NO. 21, its encoding nucleic acid is shown in SEQ ID NO. 22, and its HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NOs. 23, 24 and 25, respectively.

[0160] VH amino acid sequence:

[0161] EVQLVESGSELKKPGASVKLSCKTS GYSFTTYA ISWVRQAPGQGLEWLGR INTFTGNP TYAQGFTGRFVFSLDTSVTTAYLEISSLKAEDTAVYFC ARGASHLSGLDS WGQGGLVSVSS

[0162] VH nucleic acid sequence:

[0163] GAGGTGCAGCTGGTGGAGTCTGGGTCTGAGTTGAAGAAGCCGGGGGCCTCTGTGAAGCTTTCCTGCAAGACCTCTGGATACTCCTTCACTACTTATGCTATCAGTTGGGTGCGACAGGCCCTGGACAAGGGCTTGAGTGGCTGGGAAGGATCAACACCTTCACTGGAAACCCAACCT ATGCCCAGGGCTTCACAGGACGGTTTGTCTTCTCCTTGGACACCTCTGTCACCACGGCATATCTGGAGATCAGCAGCCTAAAGGCTGAGGACACCGCCGTCTATTTCTGTGCGAGGGGGGCGTCCCACCTAAGCGGCTTAGACTCCTGGGGCCAGGGAGGCCTGGTCAGCGTCTCCTCA

[0164] The VL amino acid sequence is shown in SEQ ID NO. 26, its encoding nucleic acid is shown in SEQ ID NO. 27, and its LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NOs. 28, 29 and 30, respectively.

[0165] VH amino acid sequence:

[0166] DIQLTQSPSSLSASVGDRVTITCRAS HNIGNW LAWYQQKPGQAPNLLIF KAS NLEYGVPSRFSGSGSGTEFTLTISSLQPDDFATYYC QQSETYPWT FGQGTRVEVK

[0167] VL nucleic acid sequence:

[0168] GACATCCAGTTGACCCAGTCTCCTTCCTCCCTGTCTGCATCTGTTGGAGACAGAGTCACCATCACTTGCCGGGCCAGTCACAACATTGGTAACTGGTTGGCCTGGTATCAGCAGAAACCAGGGCAAGCCCCTAACCTCCTCATCTTTAAGGCGTCTAATT TAGAATATGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCGACTTATTACTGCCAACAGTCTGAGACTTATCCGTGGACGTTCGGCCAAGGGACCAGGGTGGAAGTCAAA

[0169] PC0014

[0170] The VH amino acid sequence is shown in SEQ ID NO.31, its encoding nucleic acid is shown in SEQ ID NO.32, and its HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NOs.33, 34 and 35, respectively.

[0171] VH amino acid sequence:

[0172] QVQLQESGAGLLKPSETLSLTCAIY GGSFGNNY WNWIRQPPGEGLEWIGE INHRGST NSNPSLKSRVTMSVDTSKNQFSLKLTSVTAADAAVYFC ARREQLLLPDVFDI WGLGTRVAVSS

[0173] VH nucleic acid sequence:

[0174] CAGGGTGCAGCTGCAGGAGTCGGGCGCAGGACTGTTGAAGCCTTCGGAGACCCTGTCCCTCACCTGCGCTATCTATGGTGGGTCCTTCGGTAATAACTACTGGAACTGGATCCGCCAGCCCCCAGGGGAGGGGCTGGAATGGATTGGCGAAATCAATCATCGTGGAAGCACCAACTCCAAC CCGTCCCTCCAAGAGTCGAGTCACCATGTCGGTAGACACGTCCAAGAACCAGTTCTCCCTGAAGCTGACCTCTGTGACCGCCCGGGACGCGGCTGTCTATTTCTGTGCGAGACGAGAGCAGCTCCTATTGCCTGATGTCTTTGATATCTGGGGCCTCGGGACAAGGGTCGCCGTCTCTTCC

[0175] The VL amino acid sequence is shown in SEQ ID NO. 36, its encoding nucleic acid is shown in SEQ ID NO. 37, and its LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NOs. 38, 39 and 40, respectively.

[0176] VL amino acid sequence:

[0177] DIQLTQSPSSLSASVGDSVTITCRAS QRMSSY LNWYQQKPGKAPNLLIY AAS SLHSGVPSRFSGSGSGTDFTLTIASLQPEDFATYYC QQSYSAPYT FGQGTKLEIK

[0178] VL nucleic acid sequence:

[0179] GACATCCAGTTGACCCAGTCTCCGTCCTCCCTGTCTGCATCTGTAGGAGACAGTGTCACCATCACTTGCCGGGCAAGTCAGAGAATGAGCAGCTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAACCTCCTGATCTATGCTGCATCCAGTT TGCATAGTGGAGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCGCCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAGAGTTACAGTGCCCCGTACACTTTTGGCCAGGGGACCAAACTGGAGATCAAA

[0180] PC0031

[0181] The VH amino acid sequence is shown in SEQ ID NO.41, its encoding nucleic acid is shown in SEQ ID NO.42, and its HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NOs.43, 44 and 45, respectively.

[0182] VH amino acid sequence:

[0183] EVQLVESGGGVVQPGRSLRLSCAAS GFSFTKYA MHWVRQAPGKGLEWVAV IRSDGINK YYGDSVKGRFTISRDNSKSTVDLQMLSLRGEDTAVYYC AKGEGYTDYSTMYYYNGMDV WGQGTTVRVSS

[0184] VH nucleic acid sequence:

[0185] GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCTTGTGCAGCGTCTGGATTTAGTTTCACAAAATATGCAATGCACTGGGTCCGCCAGGCCCCAGGCAAGGGGCTGGAATGGGTGGCAGTTATTCGGAGTGATGGAATTAATAAATATTATGGAGACTCCG TGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAAAGCACAGTGGACCTGCAAATGCTCAGCCTGAGAGGCGAAGACACGGCTGTGTATTACTGTGCGAAAGGGGAGGGCTACACTGACTACTCCACCATGTACTATTACAATGGAATGGACGTCTGGGGCCAGGGGACCACGGTCAGAGTCTCCTCA

[0186] The VL amino acid sequence is shown in SEQ ID NO.46, its encoding nucleic acid is shown in SEQ ID NO.47, and its LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NOs.48, 49 and 50, respectively.

[0187] VL amino acid sequence:

[0188] QAVLTQPPSLSVSPGQTARISCSAR ELPNQY SHWYQQRPGQAPVLLIF KDT ERPPGIPERFSGSSSGTTVTLTISRIQPDDEADYYC QSSDNDGTHWV FGGGTHLTVRS

[0189] VL nucleic acid sequence:

[0190] CAGGCTGTGCTGACTCAGCCACCCTCGCTGTCAGTGTCCCCAGGCCAGACGGCCAGGATCTCCTGCTCTGCACGTGAATTGCCAAACCAATATTCTCATTGGTACCAGCAGAGGCCAGGCCAGGCCCCTGTATTGTTGATTTTCAAAGACACTGAGAGGCCCC CAGGCATCCCCGAGCGATTCTCTGGCTCCAGCTCAGGAACAACAGTCACGTTGACCATCTCTAGAATCCAACCAGACGACGAGGCTGACTATTATTGTCAATCATCAGACAACGATGGTACCCACTGGGTCTTCGGCGGGGGGACGCACTTAACCGTCCGCAGT

[0191] PC0034

[0192] The VH amino acid sequence is shown in SEQ ID NO.51, its encoding nucleic acid is shown in SEQ ID NO.52, and its HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NOs.53, 54 and 55, respectively.

[0193] VH amino acid sequence:

[0194] QVQLVQSGGGLVQPGGSLRLSCAAS GFTFSSYA MIWVRQAPGKGLEWVSG ISGRASPT YYADSVKGRFTISRDNSKSTLYLQMNSLRAEDTAVYYC VKDYSGSDYDILPGITALDF WGRGTLVTVSS

[0195] VH nucleic acid sequence:

[0196] CAGGTGCAGCTGGTGCAGTCTGGGGGAGGGTTGGTGCAGCCGGGGGGGTCCCTGAGACTCTCCTGTGCGGCCTCTGGATTCACGTTTAGCAGCTATGCCATGATCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGGTATTAGTGGTCGTGCAAGTCCCACATACTACGCAGACTCCG TAAAGGGCCGGTTTACCATCTCCAGAGACAATTCCAAGAGCACCCTGTATTTGCAAATGAACAGCCTGAGAGCTGAGGATACGGCCGTTTATTATTGTGTGAAAGATTATAGCGGCTCGGATTACGATATTTTGCCTGGAATCACCGCCCTTGACTTCTGGGGCCGGGGAACCCTGGTCACCGTGTCCTCA

[0197] The VL amino acid sequence is shown in SEQ ID NO.56, its encoding nucleic acid is shown in SEQ ID NO.57, and its LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NOs.58, 59 and 60, respectively.

[0198] VL amino acid sequence:

[0199] QSALTQPPSVSAAPGETARITCGGK NIGSKS VHWYQQKPGQAPVLVIH YDT DRPSGIPERFSGSNSGNTATLTISTVSAGDEADYYC QVWDSGSDHVV FGGGTKLTVL

[0200] VL nucleic acid sequence:

[0201] CAGTCTGCCCTGACTCAGCCACCCTCAGTGTCAGCGGCCCCAGGAGAGACGGCCAGGATTACCTGTGGGGGAAAGAACATTGGAAGTAAAAGTGTTCACTGGTACCAGCAGAAGCCAGGCCAGGCCCCTGTACTGGTCATCCATTATGATACCGACCGGCCC TCAGGGATCCCTGAGCGATTCTCCGGCTCCAACTCTGGGAACACGGCCACCTTGACCATCAGCACGGTCTCAGCCGGGGATGAGGCCGACTATTACTGTCAGGTGTGGGATTCAGGTAGTGATCATGTGGTTTTCGGCGGAGGGACCAAGCTGACCGTCCTA

[0202] PC0035

[0203] The VH amino acid sequence is shown in SEQ ID NO.61, its encoding nucleic acid is shown in SEQ ID NO.62, and its HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NOs.63, 64 and 65, respectively.

[0204] VH amino acid sequence:

[0205] QVQLVQSGSELMKPGASVKVSCKAS GYTFSYYA INWVRQVPGQGLEWMGW INTNTGKP SYARGLTGRFVFSLDTSVNTAFLQISSLLPDDSAIYYC ARGNLVRSLRGATGRNWIDP WGLGTLVTVSS

[0206] VH nucleic acid sequence:

[0207] CAGGTGCAGCTGGTGCAGTCTGGGTCTGAGTTGATGAAGCCTGGGGCCTCAGTGAAGGTTTCCTGCAAGGCCTCTGGATACACTTTTAGTTATTATGCTATAAATTGGGTGCGACAGGTCCCTGGACAAGGACTTGAGTGGATGGGATGGATCAACACCAACACTGGGAAACCAAGTTATGCCCGGGGCC TCACAGGACGATTTGTCTTCTCCTTGGACACGTCTGTCAACACGGCTTTTCTGCAGATCAGTAGCCTATTGCCTGACGACTCTGCCATTTATTACTGTGCGCGGGGTAATTTGGTTCGTTCGCTTCGGGGAGCCACGGGGCGCAACTGGATCGACCCCTGGGGCCTGGGAACTCTGGTCACCGTCTCCTCA

[0208] The VL amino acid sequence is shown in SEQ ID NO.66, its encoding nucleic acid is shown in SEQ ID NO.67, and its LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NOs.68, 69 and 70, respectively.

[0209] VL amino acid sequence:

[0210] QSALTQPASVSGSPGESITVSCTGS TSDVGGYNY VSWYQQHPGKAPKLLIY DVS HRPAGVSSRFSASKSGNTASLTISWLQADDEGDYYC SSYTSSNSYV FGTGTSVTVL

[0211] VL nucleic acid sequence:

[0212] CAGTCTGCCCTGACTCAGCCTGCCTCCGTGTCTGGGTCTCCTGGAGAGTCGATCACCGTCTCCTGCACTGGAAGCACCAGTGACGTTGGTGGATACAACTATGTCTCCTGGTACCAGCAACACCCAGGCAAAGCCCCCAAACTCTTGATCTATGATGTCAGTCAT CGGCCCGCAGGAGTTTCTAGTCGCTTCTCTGCGTCCAAGTCTGGCAACACGGCCTCCCTGACCATCTCTTGGCTCCAGGCTGACGACGAGGGTGATTATTACTGCAGCTCATATACAAGCAGCAATTCCTATGTCTTCGGCACTGGGACTTCGGTCACCGTCCTG

[0213] PC0037

[0214] The VH amino acid sequence is shown in SEQ ID NO.71, its encoding nucleic acid is shown in SEQ ID NO.72, and its HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NOs.73, 74 and 75, respectively.

[0215] VH amino acid sequence:

[0216] QVQLVQSGAEVKKPGESLKISCKGY GYNFATNW IGWVRQVPGKGLEWMGI IFPADSDT RYSPSFQGQVTISADKSTATAYLQWRGLKASDTAVYYC AKQSIPGWRWLDS WGQGALVTVSS

[0217] VH nucleic acid sequence:

[0218] CAGGTGCAGCTGGTGCAGTCTGGGGCAGAGGTGAAAAAGCCCGGGGAGTCCCTGAAGATCTCTTGTAAGGGTTATGGATACAACTTTGCCACGAACTGGATCGGCTGGGTGCGCCAGGTGCCCGGGAAAGGCCTGGAGTGGATGGG GATCATATTTCCTGCTGACTCTGACACCAGATATAGTCCGTCCTTCCAAGGCCAGGTCACCATATCAGCCGACAAGTCAACCGCCACCGCCTACCTCCAGTGGCGTGGCCTGAAGGCCTCGGACACCGCCGTGTATTATTGTGCGA AACAGTCAATACCTGGATGGAGGTGGCTTGACTCATGGGGCCAGGGGGCCCTGGTCACCGTCTCCTCA

[0219] The VL amino acid sequence is shown in SEQ ID NO.76, its encoding nucleic acid is shown in SEQ ID NO.77, and its LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NOs.78, 79 and 80, respectively.

[0220] VL amino acid sequence:

[0221] SYELTQPPSVSVSAGQTARITCSGD ALPKQH AHWYHQKPGQAPVLVMY KET ERPSGIPERFSGSSSGTTWTLTISAVRAEDEGDYYC QSEDSSATYLI FGGGTTLTVV

[0222] VL nucleic acid sequence:

[0223] TCTTATGAGCTGACTCAGCCACCCTCGGTGTCAGTGTCCGCAGGACAGACGGCCAGGATCACCTGCTCTGGAGATGCATTGCCAAAGCAACATGCTCATTGGTATCATCAGAAGCCAGGCCAGGCCCCTGTGTTGGTGATGTATAAAGAGACTGAGAGGCCG TCAGGGATACCTGAGCGATTCTCTGGCTCCAGTTCAGGGACAACAGTCACGTTGACAATCAGCGCAGTCCGGGCAGAGGACGAGGGCGACTATTACTGTCAATCAGAAGACAGCAGCGCCACTTATCTGATTTTTGGCGGAGGGACCACGCTGACCGTCGTA

[0224] The above eight antibodies are all of IgG1 subtype, the amino acid sequence of the heavy chain constant region is shown in SEQ ID NO.81, and the encoding nucleic acid is shown in SEQ ID NO.82.

[0225] Heavy chain constant region amino acid sequence:

[0226] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0227] Heavy chain constant region nucleic acid sequence:

[0228] GCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCCCCGGGTAAA

[0229] Among the above 8 antibodies, the light chains of PC0004, PC0010, PC0012 and PC0014 are κ light chains, the amino acid sequence of the κ light chain constant region is shown in SEQ ID NO.89, and the encoding nucleic acid thereof is shown in SEQ ID NO.90.

[0230] κ light chain constant region amino acid sequence

[0231] RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0232] κ light chain constant region nucleic acid sequence

[0233] CGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGG AGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT

[0234] Among the above 8 antibodies, the light chains of PC0031, PC0034, PC0035 and PC0037 are λ light chains, the amino acid sequence of the λ light chain constant region is shown in SEQ ID NO.91, and the encoding nucleic acid thereof is shown in SEQ ID NO.92.

[0235] The amino acid sequence of the lambda light chain constant region is as follows:

[0236] GQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS

[0237] The nucleotide sequence of the lambda light chain constant region is as follows:

[0238] GGTCAGCCCAAGGCTGCCCTCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTG GAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATGTTCA

[0239] Example 3 Reactivity of Antibodies Binding to CMV Trimer and Pentamer Complexes

[0240] In order to determine the protein components of the CMV pentamer complex recognized by the isolated pentamer-reactive antibodies, this example produced and purified recombinant CMV trimer and pentamer complexes ( Figure 1 A) and validated with known CMV gH / gL reactivity and pentamer-specific neutralizing antibodies ( Figure 3 ). Using 8I21 (gene synthesis, sequence from PDB database 5VOC) and 9I6 (gene synthesis, sequence from PDB database 5VOD) as CMV pentamer-specific antibodies and MSL-109 antibody (gene synthesis, sequence from PDB database 4LRI) as a control, the binding of 8 purified antibodies to recombinant CMV trimer and pentamer complexes was tested in ELISA. The results showed that the 8 antibodies tested could be divided into 2 groups based on their reactivity to CMV trimers and pentamers. The antibodies PC0004, PC0010, PC0012, PC0014, PC0035 and PC0037 in the first group and the control antibody MSL-109 can bind to both pentamers and trimers, but PC0004 has a weaker binding ability to CMV trimers ( Figure 3 MSL-109 binds to the gH / gL subunits of CMV trimer and pentamer complexes. PC0031 and PC0034 in group 2 and control antibodies 8I21 and 9I6 can only bind to CMV pentamers ( Figure 3 Since gH / gL are present in both CMV trimer and pentameric complexes, these results suggest that the antibodies in group 1 may recognize gH / gL in the complex, while the two antibodies in group 2 may recognize pUL128 / 130 / 131A ( Figure 3 ), the two groups of antibodies recognize different subunits or different antigenic epitopes, indicating that the two groups of antibodies have different antiviral activities. This has important clinical significance for developing an antibody that fully targets the binding region of the pentamer to the cell receptor and completely blocks the binding of the virus to cells.

[0241] Example 4 Neutralizing Activity of Antibodies Specific to CMV Pentamers

[0242] All eight purified antibodies were tested for their ability to neutralize CMV in human embryonic lung fibroblasts (MRC-5, purchased from ATCC, catalog number: CCL-171). The results showed that three of the six antibodies in group 1, PC0012, PC0014, and PC0035, along with the control antibody MSL-109, neutralized the HCMV laboratory standard Towne strain with EC50s of 0.648-0.938 μg / mL, and also neutralized the HCMV wild-type strain BE13 / 2012 with EC50s of 0.299-1.480 μg / mL ( Figure 4 A and 4B). In the fibroblast-based neutralization assay, two pentamer-specific antibodies in group 2, PC0031 and PC0034, did not neutralize CMV Towne strain ( Figure 4 A), nor did it neutralize the clinical isolate BE13 / 2012 (data not shown), which is consistent with the performance of the known pentamer-specific neutralizing antibodies 8I21 and 9I6 (39). However, among the two pentamer-specific antibodies, only PC0034 was shown to neutralize the two tested HCMV wild-type strains in the ARPE-19 epithelial cell-based neutralization assay, including VR1814 (GenBank: GU179289.1) (40), NR (GenBank: KX544831.1) (40) and the recombinant HCMV strain AD169 FIX (chimeric expression of the complete pentamer membrane protein on the standard strain AD169 (GenBank: FJ527563.1)) (40), with EC50s of 0.068, 0.070 and 0.079 μg / ml, respectively ( Figure 4 C).

[0243] Example 5 Antibody Binding Ability and Mechanism of Neutralizing CMV

[0244] (1) Binding and cross-reactivity of neutralizing antibodies

[0245] The binding affinities of these two groups of HCMV neutralizing antibodies to purified gH / gL / gO trimers and gH / gL / UL128 / UL130 / UL131A pentamers were determined by SPR and compared with the known neutralizing antibody MSL-109. The three gH / gL-binding neutralizing antibodies PC0012, PC0014, and PC0035 had high affinity for pentamers, with KD of 2.85×10 -10 M, 3.23×10 -10 M and 1.83×10 -10 M( Figure 5 A), the affinity KD for binding to trimer was 9.38×10 -11 M, 1.00×10 -9 M and 1.01×10 -10 M( Figure 5 B). The binding affinity of antibodies PC0012, PC0014, and PC0035 to the pentamer is approximately 4-7 times higher than that of antibody MSL-109 ( Figure 5 A and Table 2), and the affinity to the trimer is about 20-230 times higher than that of antibody MSL-109 ( Figure 5 B and Table 2).

[0246] Table 2. Affinity of antibodies PC0012, PC0014 and PC0035 binding to HCMV pentamers and trimers

[0247]

[0248] To gain further insights into the relationship between the epitopes recognized by antibodies PC0012, PC0014, and PC0035 and the known neutralizing antibody MSL-109, the cross-competitive blocking binding of these antibodies to CMV pentamers was determined by SPR and ELISA ( Figure 6 The results showed that the antibodies in group 1 did not compete with the antibodies in group 2 for binding, and vice versa in ELISA tests ( Figure 6 Among the three newly isolated gH / gL binding neutralizing antibodies in the first group, PC0012 blocked the binding of antibodies PC0014 and PC0035 to CMV pentamers, while PC0014 only partially blocked the binding, and PC0035 did not block the binding of PC0012 to CMV pentamers at all. PC0014 and PC0035 did not block each other's binding to CMV pentamers ( Figure 6 PC0012 and PC0035 completely blocked the binding of MSL-109 to CMV pentamers, while MSL-109 did not block the binding of PC0012 to CMV pentamers, but could block the binding of PC0035 to CMV pentamers. PC0014 and MSL-109 did not block each other's binding to CMV pentamers ( Figure 6 ).

[0249] Previous studies have shown that in MSL-109-resistant viral mutants, the W168C / R, P171H / S, and D446N mutations of the gH subunit lead to viral tolerance to the antibody MSL-109, while the antibody MSL-109 does not bind to the gH / gL with the W168C / R mutation (41). In order to evaluate whether the binding of antibodies PC0012, PC0014, and PC0035 to pentamers is affected by mutations at these three sites on the gH subunit, pentamer mutants targeting these three sites were recombinantly expressed. The experimental results showed that antibodies PC0012, PC0014, and PC0035 strongly bound to these three pentamer mutants ( Figure 7 A), while the antibody MSL-109 did not bind to these mutants or bound only weakly ( Figure 7 A). Therefore, antibodies PC0012, PC0014, and PC0035 recognize antigenic epitopes in similar regions, and their epitopes are different from the epitope of antibody MSL-109, but may partially overlap or be close to it.

[0250] (2) Mechanism of neutralization of CMV by gH / gL-binding antibodies

[0251] It has been reported that neutralizing antibodies that bind gH / gL and gB exert a neutralizing effect after the virus adheres to the host cell (35,41). To clarify how antibodies PC0012, PC0014, and PC0035 affect virus-infected cells, the complexes of antibodies PC0012, PC0014, and PC0035 with the virus were first incubated in 4°C pre-cooled MRC-5 cells. The results showed that antibodies PC0012, PC0014, and PC0035, as well as antibodies MSL-109 and unrelated antibody TRN006 (patent: CN103910796B), did not reduce the amount of CMV virus on the cell membrane surface, while the positive control heparin could hardly detect any viral DNA copies ( Figure 7 B). Subsequently, the researchers investigated whether antibodies PC0012, PC0014, and PC0035 could interfere with viral infection of cells attached to the cell membrane. CMV virus was pre-inoculated into MRC-5 cells at 4°C and incubated for 30 minutes before the addition of antibodies PC0012, PC0014, and PC0035. The results showed that antibodies PC0012, PC0014, and PC0035, as well as antibody MSL-109, were able to block CMV infection of cells ( Figure 7 C) Therefore, antibodies PC0012, PC0014, and PC0035, as well as antibody MSL-109, do not block viral adhesion to cells, but rather block the viral invasion process after adhesion.

[0252] HCMV also spreads between cells by mediating syncytia formation (42). The inventors evaluated the effects of antibodies PC0012, PC0014, and PC0035 on the spread of HCMV between cells. Five days after virus inoculation, the expression of CMV virus-specific CMV IE1 / IE2 proteins was detected to determine the status of HCMV infection. Compared with the larger and more numerous brown-stained satellite replication centers seen in virus-infected cells without added antibodies (only virus) or treated with the negative control antibody TRN006, uniformly dispersed smaller sizes and fewer brown-stained satellite virus replication centers were observed in infected cells treated with antibodies PC0012, PC0014, PC0035, or MSL-109 ( Figure 7 D) These results indicate that antibodies PC0012, PC0014, and PC0035, as well as MSL-109, are able to inhibit the cell-to-cell spread of HCMV in fibroblasts.

[0253] Example 6 Antibody PC0034 blocks the binding of HCMV pentamers to epithelial and endothelial cells

[0254] The CMV neutralizing antibody 9I6 was specifically defined to recognize a pentamer-specific site and block the binding of CMV pentamers to the cell surface receptor Nrp2 (43). The affinity of antibody PC0034 for binding to CMV pentamers was 1.05×10 -10 M, while antibody 9I6 had a slightly lower affinity for pentamer binding, with a KD of 4.84×10 -10 M, lower than PC0034 ( Figure 8 A). SPR analysis showed that antibodies PC0034 and 9I6 could completely block each other's binding to CMV pentamer ( Figure 8 B). Recombinantly expressed HCMV pentamer protein can bind to epithelial cells (APRE-19, purchased from ATCC, catalog number: CRL-2302) and endothelial cells (HUVEC, purchased from ATCC, catalog number: CRL-1730), and there is a dose-response relationship ( Figure 8 C). After antibody PC0034 binds to HCMV pentamer protein, antibody PC0034 can inhibit the binding of pentamer to epithelial cells and endothelial cells ( Figure 8 D) Therefore, these data indicate that antibody PC0034 exerts its antiviral activity by blocking the binding of pentamers to cell surface receptors.

[0255] Example 7 Study on the epitope recognized by antibody PC0034

[0256] Figure 9The binding results of antibody PC0034 to HCMV pentamer mutants are shown. The affinity changes of antibody PC0034 to these four pentamer mutants were evaluated by SPR, among which UL131A_E23A decreased by 9.96 times, UL131A_K27A decreased by 33.93 times, UL128_K47A decreased by 9.33 times, and UL128_T94A decreased by 81.06 times ( Figure 9 A to 9D and Table 3). The affinity of antibody PC0034 for pentamers is closely related to two sites on the pentamer (UL128_T94 and UL131A_K27), but the affinity of antibody 9I6 for pentamer mutants of these two sites does not change significantly. Therefore, the binding epitope of antibody PC0034 is different from that of 9I6. At the same time, in the experiment of pentamer protein binding to cells, it was found that the binding of pentamer mutants (UL128_T94A and UL131A_K27A) to epithelial cells was significantly reduced ( Figure 9 F and 9G). These results indicate that the binding epitope of antibody PC0034 on the pentamer overlaps with the binding epitope of the pentamer to cell surface receptors.

[0257] Table 3. Affinity of antibodies binding to HCMV pentamer and pentamer mutants

[0258]

[0259] References:

[0260] 1. Adland E, Klenerman P, Goulder P, Matthews PC. 2015. Ongoing burden of disease and mortality from HIV / CMV coinfection in Africa in the antiretroviral therapy era. Front Microbiol 6:1016.

[0261] 2.Styczynski J.2018.Who Is the Patient at Risk of CMV Recurrence:AReview of the Current Scientific Evidence with a Focus on Hematopoietic CellTransplantation.Infect Dis Ther 7:1-16.

[0262] 3.Ye L,Qian Y,Yu W,Guo G,Wang H,Xue X.2020.Functional Profile ofHuman Cytomegalovirus Genes and Their Associated Diseases:A Review.FrontMicrobiol 11:2104.

[0263] 4.Boppana SB,Ross SA,Fowler KB.2013.Congenital cytomegalovirusinfection:clinical outcome.Clin Infect Dis 57 Suppl 4:S178-81.

[0264] 5.Kirby T.2016.Congenital cytomegalovirus—a neglected healthproblem.The Lancet Infectious Diseases 16:900-901.

[0265] 6.Vollmer B,Grunewald K.2020.Herpesvirus membrane fusion-a teameffort.Curr Opin Struct Biol 62:112-120.

[0266] 7.Connolly SA,Jardetzky TS,Longnecker R.2021.The structural basis ofherpesvirus entry.Nat Rev Microbiol 19:110-121.

[0267] 8.Vanarsdall AL,Ryckman BJ,Chase MC,Johnson DC.2008.Humancytomegalovirus glycoproteins gB and gH / gL mediate epithelial cell-cellfusion when expressed either in cis or in trans.J Virol 82:11837-50.

[0268] 9.Vanarsdall AL,Johnson DC.2012.Human cytomegalovirus entry intocells.Curr Opin Virol 2:37-42.

[0269] 10.Sathiyamoorthy K,Chen J,Longnecker R,Jardetzky TS.2017.TheCOMPLEXity in herpesvirus entry.Curr Opin Virol 24:97-104.

[0270] 11.Ciferri C,Chandramouli S,Donnarumma D,Nikitin PA,Cianfrocco MA,Gerrein R,Feire AL,Barnett SW,Lilja AE,Rappuoli R,Norais N,Settembre EC,CarfiA.2015.Structural and biochemical studies of HCMV gH / gL / gO and Pentamerreveal mutually exclusive cell entry complexes.Proc Natl Acad Sci U S A 112:1767-72.

[0271] 12.Li G,Nguyen CC,Ryckman BJ,Britt WJ,Kamil JP.2015.A viral regulatorof glycoprotein complexes contributes to human cytomegalovirus celltropism.Proc Natl Acad Sci U S A 112:4471-6.

[0272] 13.Luganini A,Cavaletto N,Raimondo S,Geuna S,Gribaudo G.2017.Loss ofthe Human Cytomegalovirus US16 Protein Abrogates Virus Entry into Endothelialand Epithelial Cells by Reducing the Virion Content of the Pentamer.J Virol91.

[0273] 14.Siddiquey MNA,Schultz EP,Yu Q,Amendola D,Vezzani G,Yu D,Maione D,Lanchy JM,Ryckman BJ,Merola M,Kamil JP.2021.The Human Cytomegalovirus ProteinUL116 Interacts with the Viral Endoplasmic-Reticulum-Resident GlycoproteinUL148 and Promotes the Incorporation of gH / gL Complexes into Virions.J Virol95:e0220720.

[0274] 15.Vezzani G,Amendola D,Yu D,Chandramouli S,Frigimelica E,Maione D,Merola M.2021.The Human Cytomegalovirus UL116 Glycoprotein Is a Chaperone toControl gH-Based Complexes Levels on Virions.Front Microbiol 12:630121.

[0275] 16.Hahn G, Revello MG, Patrone M, Percivalle E, Campanini G, Sarasini A, Wagner M, Gallina A, Milanesi G, Koszinowski U, Baldanti F, Gerna G leukocytes.J Virol 78:10023–33.

[0276] 17.Ryckman BJ,Jarvis MA,Drummond DD,Nelson JA,Johnson DC.2006.Humancytomegalovirus entry into epithelial and endothelial cells depends on genesUL128 to UL150 and occurs by endocytosis and low-pH fusion.J Virol 80:710-22.

[0277] 18.Jiang XJ,Adler B,Sampaio KL,Digel M,Jahn G,Ettischer N,StierhofYD,Scrivano L,Koszinowski U,Mach M,Sinzger C.2008.UL74 of human cytomegalovirus contributes to virus release by promoting secondaryenvelopment of virions.J Virol 82:2802–12.

[0278] 19.Wille PT,Knoche AJ,Nelson JA,Jarvis MA,Johnson DC.2010.A humancytomegalovirus gO-null mutant fails to incorporate gH / gL into the virionenvelope and is unable to enter fibroblasts and epithelial and endothelialcells.J Virol 84:2585-96.

[0279] 20.Zhou M,Lanchy JM,Ryckman BJ.2015.Human Cytomegalovirus gH / gL / gOPromotes the Fusion Step of Entry into All Cell Types,whereas gH / gL / UL128-131Broadens Virus Tropism through a Distinct Mechanism.J Virol 89:8999-9009.

[0280] 21.Smith MG.1956.Propagation in tissue cultures of a cytopathogenicvirus from human salivary gland virus(SGV)disease.Proc Soc Exp Biol Med 92:424-30.

[0281] 22.Gomes AC,Griffiths PD,Reeves MB.2019.The Humoral Immune ResponseAgainst the gB Vaccine:Lessons Learnt from Protection in Solid OrganTransplantation.Vaccines(Basel)7.

[0282] 23.Snydman DR.1990.Cytomegalovirus immunoglobulins in the preventionand treatment of cytomegalovirus disease.Rev Infect Dis 12Suppl 7:S839-48.

[0283] 24.Hakki M.2020.Moving Past Ganciclovir and Foscarnet:Advances in CMVTherapy.Curr Hematol Malig Rep 15:90-102.

[0284] 25.Imlay HN,Kaul DR.2021.Letermovir and Maribavir for the Treatmentand Prevention of Cytomegalovirus Infection in Solid Organ and Stem CellTransplant Recipients.Clin Infect Dis 73:156-160.

[0285] 26.Bonaros N,Mayer B,Schachner T,Laufer G,Kocher A.2008.CMV-hyperimmune globulin for preventing cytomegalovirus infection and disease insolid organ transplant recipients:a meta-analysis.Clin Transplant 22:89-97.

[0286] 27.Alsuliman T,Kitel C,Dulery R,Guillaume T,Larosa F,Cornillon J,Labussiere-Wallet H,Mediavilla C,Belaiche S,Delage J,Alain S,Yakoub-AghaI.2018.Cytotect(R)CP as salvage therapy in patients with CMV infectionfollowing allogeneic hematopoietic cell transplantation:a multicenterretrospective study.Bone Marrow Transplant 53:1328-1335.

[0287] 28.Jenks JA,Goodwin ML,Permar SR.2019.The Roles of Host and ViralAntibody Fc Receptors in Herpes Simplex Virus(HSV)and Human Cytomegalovirus(HCMV)Infections and Immunity.Front Immunol 10:2110.

[0288] 29.Kagan KO,Enders M,Schampera MS,Baeumel E,Hoopmann M,Geipel A,BergC,Goelz R,De Catte L,Wallwiener D,Brucker S,Adler SP,Jahn G,HamprechtK.2019.Prevention of maternal-fetal transmission of cytomegalovirus afterprimary maternal infection in the first trimester by biweeklyhyperimmunoglobulin administration.Ultrasound Obstet Gynecol 53:383-389.

[0289] 30.Chou S,Marousek G,Li S,Weinberg A.2008.Contrasting drug resistancephenotypes resulting from cytomegalovirus DNA polymerase mutations at thesame exonuclease locus.J Clin Virol 43:107-9.

[0290] 31.Ohlin M,Soderberg-Naucler C.2015.Human antibody technology and thedevelopment of antibodies against cytomegalovirus.Mol Immunol 67:153-70.

[0291] 32.Tabata T,Petitt M,Fang-Hoover J,Freed DC,Li F,An Z,Wang D,Fu TM,Pereira L.2019.Neutralizing Monoclonal Antibodies Reduce HumanCytomegalovirus Infection and Spread in Developing Placentas.Vaccines(Basel)7.

[0292] 33.Lilleri D,Kabanova A,Revello MG,Percivalle E,Sarasini A,Genini E,Sallusto F,Lanzavecchia A,Corti D,Gerna G.2013.Fetal human cytomegalovirustransmission correlates with delayed maternal antibodies to gH / gL / pUL128-130-131 complex during primary infection.PLoS One 8:e59863.

[0293] 34.Blanco-Lobo P,Cordero E,Martin-Gandul C,Gentil MA,Suarez-ArtachoG,Sobrino M,Aznar J,Perez-Romero P.2016.Use of antibodies neutralizingepithelial cell infection to diagnose patients at risk for CMV Disease aftertransplantation.J Infect 72:597-607.

[0294] 35.Gardner TJ,Stein KR,Duty JA,Schwarz TM,Noriega VM,Kraus T,MoranTM,Tortorella D.2016.Functional screening for anti-CMV biologics identifies abroadly neutralizing epitope of an essential envelope protein.Nat Commun 7:13627.

[0295] 36.Ishida JH,Patel A,Mehta AK,Gatault P,McBride JM,Burgess T,DerbyMA,Snydman DR,Emu B,Feierbach B,Fouts AE,Maia M,Deng R,Rosenberger CM,GennaroLA,Striano NS,Liao XC,Tavel JA.2017.Phase 2 Randomized,Double-Blind,Placebo-Controlled Trial of RG7667,a Combination Monoclonal Antibody,for Preventionof Cytomegalovirus Infection in High-Risk Kidney TransplantRecipients.Antimicrob Agents Chemother 61.

[0296] 37.Eisenberg,R.J.,Cairns,T.M.&Cohen,G.H.HCMV grabs a mechanism toescape neutralization.Cell hostµbe 10,177-178,doi:10.1016 / j.chom.2011.08.011(2011).

[0297] 38.Tiller T,Meffre E,Yurasov S,Tsuiji M,Nussenzweig MC,WardemannH.2008.Efficient generation of monoclonal antibodies from single human Bcells by single cell RT-PCR and expression vector cloning.J Immunol Methods329:112-24.

[0298] 39.Macagno A,Bernasconi NL,Vanzetta F,Dander E,Sarasini A,Revello MG,Gerna G,Sallusto F,Lanzavecchia A.2010.Isolation of human monoclonalantibodies that potently neutralize human cytomegalovirus infection bytargeting different epitopes on the gH / gL / UL128-131A complex.J Virol 84:1005-13.

[0299] 40.Ye X,Su H,Wrapp D,Freed DC,Li F,Yuan Z,Tang A,Li L,Ku Z,Xiong W,Jaijyan D,Zhu H,Wang D,McLellan JS,Zhang N,Fu TM,An Z.2020.Recognition of ahighly conserved glycoprotein B epitope by a bivalent antibody neutralizingHCMV at a post-attachment step.PLoS Pathog 16:e1008736.

[0300] 41.Fouts AE,Comps-Agrar L,Stengel KF,Ellerman D,Schoeffler AJ,WarmingS,Eaton DL,Feierbach B.2014.Mechanism for neutralizing activity by the anti-CMV gH / gL monoclonal antibody MSL-109.Proc Natl Acad Sci U S A 111:8209-14.

[0301] 42.Ye X,Su H,Wrapp D,Freed DC,Li F,Yuan Z,Tang A,Li L,Ku Z,Xiong W,Jaijyan D,Zhu H,Wang D,McLellan JS,Zhang N,Fu TM,An Z.2020.Recognition of ahighly conserved glycoprotein B epitope by a bivalent antibody neutralizingHCMV at a post-attachment step.PLoS Pathog 16:e1008736.

[0302] 43. Martinez-Martin N, Marcandalli J, Huang CS, Arthur CP, Perotti M, Foglierini M, Ho H, Dosey AM, Shriver S, Payandeh J, Leitner A, Lanzavecchia A, Perez L, Ciferri C. 2018. An Unbiased Screen for Human Cytomegalovirus Identifies Neuropilin-2 as a Central Viral Receptor. Cell 174:1158-1171 e19.

Claims

1. An anti-human cytomegalovirus antibody or antigen-binding fragment thereof, wherein the CDR combinations of the heavy and light chain variable regions are as follows: The heavy chain CDR1, CDR2 and CDR3 sequences are SEQ ID NOs. 43-45, and the light chain CDR1, CDR2 and CDR3 sequences are SEQ ID NOs. 48-50.

2. The antibody or antigen-binding fragment thereof according to claim 1, which has the following combination of heavy chain variable region and light chain variable region: The heavy chain variable region comprises or consists of an amino acid sequence that is at least 90% identical to the amino acid sequence shown in SEQ ID NO.41, and the light chain variable region comprises or consists of an amino acid sequence that is at least 90% identical to the amino acid sequence shown in SEQ ID NO.

46.

3. The antibody or antigen-binding fragment thereof according to claim 1, which has the following combination of heavy chain variable region and light chain variable region: The heavy chain variable region has the amino acid sequence shown in SEQ ID NO.41, and the light chain variable region has the amino acid sequence shown in SEQ ID NO.

46.

4. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 3, wherein the antibody is an IgG1, IgG2, IgG3 or IgG4 type antibody. The antibody or antigen-binding fragment thereof according to claim 4 , wherein the antibody is an IgG1 antibody.

6. The antibody or antigen-binding fragment thereof according to claim 4, wherein the constant region of the heavy chain comprises or consists of an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO. 81; and / or The constant region of the light chain comprises or consists of an amino acid sequence that is at least 90% identical to the amino acid sequence shown in SEQ ID NO. 89 or 91.

7. The antibody or antigen-binding fragment thereof according to claim 4, wherein the constant region of the heavy chain has the amino acid sequence shown in SEQ ID NO. 81; and / or the constant region of the light chain has the amino acid sequence shown in SEQ ID NO. 89 or 91.

8. A nucleic acid encoding the antibody or antigen-binding fragment thereof according to any one of claims 1 to 7.

9. The nucleic acid according to claim 8, wherein the nucleic acid comprises a nucleic acid sequence encoding an antibody heavy chain variable region of SEQ ID NO. 42 or any variant thereof, and a nucleic acid sequence encoding an antibody light chain variable region of SEQ ID NO. 47 or any variant thereof.

10. A vector comprising the nucleic acid according to claim 8 or 9.

11. A host cell comprising the nucleic acid of claim 8 or 9 or the vector of claim 10.

12. A pharmaceutical composition comprising the antibody or antigen-binding fragment thereof according to any one of claims 1 to 7, the nucleic acid according to claim 8 or 9, the vector according to claim 10 and / or the cell according to claim 11.

13. An immunoconjugate comprising the antibody or antigen-binding fragment thereof according to any one of claims 1 to 7 and a label.

14. A method of producing the antibody or antigen-binding fragment thereof according to any one of claims 1 to 7, the method comprising culturing a host cell comprising the nucleic acid of claim 8 or 9 or the vector of claim 10.

15. Use of the antibody or antigen-binding fragment thereof according to any one of claims 1 to 7, the nucleic acid according to claim 8 or 9, the vector according to claim 10, the host cell according to claim 11, or the immunoconjugate according to claim 13 for preparing a pharmaceutical composition or kit for detecting, treating, preventing and / or alleviating CMV infection or CMV-related diseases.

16. The use according to claim 15, wherein the CMV is HCMV.

17. Use of the antibody or antigen-binding fragment thereof according to any one of claims 1 to 7, the nucleic acid according to claim 8 or 9, the vector according to claim 10, the host cell according to claim 11, the pharmaceutical composition according to claim 12 or the immunoconjugate according to claim 13 in the preparation of a medicament for improving, enhancing or stimulating resistance in a human subject infected with HCMV.

18. A method for neutralizing HCMV in an individual or sample for non-diagnostic and non-therapeutic purposes, comprising contacting the antibody or antigen-binding fragment thereof according to any one of claims 1 to 7 with the individual or sample, and testing the ability of the antibody or antigen-binding fragment thereof to bind to neutralize HCMV.

Images