Specific monoclonal antibody for resisting Nipah virus Post-F protein

Abstract

The invention discloses a specific monoclonal antibody for resisting a Nipah virus Post-F protein, the monoclonal antibody comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3; wherein the HCDR1, the HCDR2 and the HCDR3 respectively comprise the HCDR1, the HCDR2 and the HCDR3 in a heavy chain variable region of which the amino acid sequence is any one of SEQ ID NO: 4, 13, 20, 25, 30, 34 or 42, or respectively have at least 85% sequence identity with the HCDR1, the HCDR2 and the HCDR3; and / or the LCDR1, the LCDR2 and the LCDR3 respectively comprise LCDR1, LCDR2 and LCDR3 in a light chain variable region with the amino acid sequence of any one of SEQ ID NO: 9, 16, 23, 28, 32, 37 or 46, or respectively have at least 85% sequence identity with the LCDR1, the LCDR2 and the LCDR3. The anti-Nipah virus Post-F protein monoclonal antibody disclosed by the invention can be specifically combined with a Post-F conformation protein, and has the application potential of being developed into a Nipah virus antibody detection kit or a virus neutralizing antibody drug.

Description

Technical Field

[0001] The invention provides a specific monoclonal antibody for Nipah virus Post-F protein. Background Art

[0002] Nipah virus (Niv) is a new type of zoonotic virus belonging to the family Paramyxoviridae. The viral genome consists of a non-segmented single-stranded negative-strand RNA of approximately 18.2 kb in length, encoding six proteins: nucleocapsid (N), phosphoprotein (P), matrix protein (M), fusion protein (F), glycoprotein (G) and RNA polymerase (L). Glycoprotein G recognizes and binds to Ephrin-B2 and B3 receptors on the surface of host cells, and fusion protein F induces fusion of the virus with the host cell membrane, thereby facilitating viral invasion. Nipah virus can cause widespread vasculitis, and infected people have symptoms such as fever, severe headache, and inflammatory membrane inflammation, which can cause serious harm to humans and animals. Developing antibodies against the fusion site of the F protein can effectively protect animals from infection. Post-F uses protein antigens to immunize mice, and selects mice with high antibody titers for immune library construction for screening mouse monoclonal antibodies.

[0003] Therefore, it is of great significance to develop antibodies against Nipah Post-F by immunizing against Post-F protein. Summary of the invention

[0004] The present invention provides a specific antibody against Nipah virus Post-F protein, which is of great significance for the treatment and prevention of subsequent Nipah virus infection.

[0005] In a first aspect of the present invention, an antibody or an antigen-binding fragment thereof against Nipah virus Post-F protein is provided, wherein the antibody or the antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3;

[0006] wherein the HCDR1, HCDR2 and HCDR3 respectively comprise the HCDR1, HCDR2 and HCDR3 in the heavy chain variable region of any one of SEQ ID NO: 4, 13, 20, 25, 30, 34 or 42, or respectively have at least 85% sequence identity thereto; and

[0007] The LCDR1, LCDR2 and LCDR3 respectively comprise LCDR1, LCDR2 and LCDR3 in the light chain variable region of any one of SEQ ID NO: 9, 16, 23, 28, 32, 37 or 46, or have at least 85% sequence identity thereto.

[0008] In some embodiments, the HCDR1 comprises the amino acid sequence shown in SEQ ID NO:1, 11, 18 or 39, the HCDR2 comprises the amino acid sequence shown in SEQ ID NO:2, 12 or 40, the HCDR3 comprises the amino acid sequence shown in SEQ ID NO:3, 19 or 41, the LCDR1 comprises the amino acid sequence shown in SEQ ID NO:6 or 44, the LCDR2 comprises the amino acid sequence shown in SEQ ID NO:7, and the LCDR3 comprises the amino acid sequence shown in SEQ ID NO:8, 15, 22, 27, 36 or 45.

[0009] In some embodiments, in the antibody or antigen-binding fragment thereof:

[0010] (i) the HCDR1 comprises the amino acid sequence shown in SEQ ID NO:1, the HCDR2 comprises the amino acid sequence shown in SEQ ID NO:2, the HCDR3 comprises the amino acid sequence shown in SEQ ID NO:3, the LCDR1 comprises the amino acid sequence shown in SEQ ID NO:6, the LCDR2 comprises the amino acid sequence shown in SEQ ID NO:7, and the LCDR3 comprises the amino acid sequence shown in SEQ ID NO:8; or

[0011] (ii) the HCDR1 comprises the amino acid sequence shown in SEQ ID NO:11, the HCDR2 comprises the amino acid sequence shown in SEQ ID NO:12, the HCDR3 comprises the amino acid sequence shown in SEQ ID NO:3, the LCDR1 comprises the amino acid sequence shown in SEQ ID NO:6, the LCDR2 comprises the amino acid sequence shown in SEQ ID NO:7, and the LCDR3 comprises the amino acid sequence shown in SEQ ID NO:15; or

[0012] (iii) the HCDR1 comprises the amino acid sequence shown in SEQ ID NO:18, the HCDR2 comprises the amino acid sequence shown in SEQ ID NO:2, the HCDR3 comprises the amino acid sequence shown in SEQ ID NO:19, the LCDR1 comprises the amino acid sequence shown in SEQ ID NO:6, the LCDR2 comprises the amino acid sequence shown in SEQ ID NO:7, and the LCDR3 comprises the amino acid sequence shown in SEQ ID NO:22; or

[0013] (iv) the HCDR1 comprises the amino acid sequence shown in SEQ ID NO:11, the HCDR2 comprises the amino acid sequence shown in SEQ ID NO:12, the HCDR3 comprises the amino acid sequence shown in SEQ ID NO:3, the LCDR1 comprises the amino acid sequence shown in SEQ ID NO:6, the LCDR2 comprises the amino acid sequence shown in SEQ ID NO:7, and the LCDR3 comprises the amino acid sequence shown in SEQ ID NO:27; or

[0014] (v) the HCDR1 comprises the amino acid sequence shown in SEQ ID NO:11, the HCDR2 comprises the amino acid sequence shown in SEQ ID NO:12, the HCDR3 comprises the amino acid sequence shown in SEQ ID NO:3, the LCDR1 comprises the amino acid sequence shown in SEQ ID NO:6, the LCDR2 comprises the amino acid sequence shown in SEQ ID NO:7, and the LCDR3 comprises the amino acid sequence shown in SEQ ID NO:15; or

[0015] (vi) the HCDR1 comprises the amino acid sequence shown in SEQ ID NO:1, the HCDR2 comprises the amino acid sequence shown in SEQ ID NO:2, the HCDR3 comprises the amino acid sequence shown in SEQ ID NO:3, the LCDR1 comprises the amino acid sequence shown in SEQ ID NO:6, the LCDR2 comprises the amino acid sequence shown in SEQ ID NO:7, and the LCDR3 comprises the amino acid sequence shown in SEQ ID NO:36; or

[0016] (vii) the HCDR1 comprises the amino acid sequence shown in SEQ ID NO:39, the HCDR2 comprises the amino acid sequence shown in SEQ ID NO:40, the HCDR3 comprises the amino acid sequence shown in SEQ ID NO:41, the LCDR1 comprises the amino acid sequence shown in SEQ ID NO:44, the LCDR2 comprises the amino acid sequence shown in SEQ ID NO:7, and the LCDR3 comprises the amino acid sequence shown in SEQ ID NO:45.

[0017] In some embodiments, the framework region of the heavy chain variable region and / or the light chain variable region is a human, mouse or rabbit framework region.

[0018] In some embodiments, the heavy chain variable region comprises an amino acid sequence as shown in SEQ ID NO:4, 13, 20, 25, 30, 34 or 42, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:4, 13, 20, 25, 30, 34 or 42; and / or

[0019] The light chain variable region comprises an amino acid sequence as shown in SEQ ID NO:9, 16, 23, 28, 32, 37 or 46, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:9, 16, 23, 28, 32, 37 or 46;

[0020] In some embodiments, in the antibody or antigen-binding fragment thereof:

[0021] (i) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO:4, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO:9; or

[0022] (ii) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 13, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 16; or

[0023] (iii) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 20, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 23; or

[0024] (iv) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 25, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 28; or

[0025] (v) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 30, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 32; or

[0026] (vi) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 34, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 37; or

[0027] (vii) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO:42, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO:46.

[0028] In some embodiments, the antibody or antigen-binding fragment thereof is a full-length antibody, Fab, Fab', F(ab')2 or Fv, and the Fv is preferably a scFv.

[0029] In some embodiments, the antibody is a full-length antibody.

[0030] In some embodiments, the amino acid sequence of the heavy chain of the antibody is shown in SEQ ID NO:5, 14, 21, 26, 31, 35 or 43; and / or the amino acid sequence of the light chain of the antibody is shown in SEQ ID NO:10, 17, 24, 29, 33, 38 or 47.

[0031] In some embodiments, in the antibody or antigen-binding fragment thereof:

[0032] (i) the amino acid sequence of the heavy chain of the antibody is shown in SEQ ID NO: 5, and the amino acid sequence of the light chain of the antibody is shown in SEQ ID NO: 10; or

[0033] (ii) the amino acid sequence of the heavy chain of the antibody is shown in SEQ ID NO: 14, and the amino acid sequence of the light chain of the antibody is shown in SEQ ID NO: 17; or

[0034] (iii) the amino acid sequence of the heavy chain of the antibody is shown in SEQ ID NO: 21, and the amino acid sequence of the light chain of the antibody is shown in SEQ ID NO: 24; or

[0035] (iv) the amino acid sequence of the heavy chain of the antibody is shown in SEQ ID NO: 26, and the amino acid sequence of the light chain of the antibody is shown in SEQ ID NO: 29; or

[0036] (v) the amino acid sequence of the heavy chain of the antibody is shown in SEQ ID NO:31, and the amino acid sequence of the light chain of the antibody is shown in SEQ ID NO:33; or

[0037] (vi) the amino acid sequence of the heavy chain of the antibody is shown in SEQ ID NO: 35, and the amino acid sequence of the light chain of the antibody is shown in SEQ ID NO: 38; or

[0038] (vii) the amino acid sequence of the heavy chain of the antibody is shown in SEQ ID NO:43, and the amino acid sequence of the light chain of the antibody is shown in SEQ ID NO:47.

[0039] In a second aspect of the present invention, an isolated nucleic acid is provided, wherein the isolated nucleic acid encodes the antibody or antigen-binding fragment thereof as described in the first aspect of the present invention.

[0040] In some embodiments, the nucleotide sequence encoding the heavy chain of the antibody or its antigen-binding fragment is as shown in SEQ ID NO:52, 62, 69, 75, 80, 84 or 92; and / or, the nucleotide sequence encoding the light chain of the antibody or its antigen-binding fragment is as shown in SEQ ID NO:57, 65, 73, 78, 82, 87 or 96.

[0041] In the third aspect of the present invention, a recombinant expression vector is provided, wherein the recombinant expression vector comprises the isolated nucleic acid as described in the second aspect of the present invention.

[0042] In some embodiments, the recombinant expression vector is a plasmid, a cosmid, a phage or a viral vector, and the viral vector is preferably a retroviral vector, a lentiviral vector, an adenoviral vector or an adeno-associated viral vector.

[0043] In the fourth aspect of the present invention, a transformant is provided, the transformant comprising the isolated nucleic acid as described in the second aspect of the present invention or the recombinant expression vector as described in the third aspect of the present invention, and the host cell of the transformant is a prokaryotic cell or a eukaryotic cell.

[0044] In some embodiments, the host cell is a yeast cell or a mammalian cell, such as a HEK293 cell or a CHO cell.

[0045] In some embodiments, the host cell is a 293F cell.

[0046] In the fifth aspect of the present invention, a method for preparing an antibody or an antigen-binding fragment thereof against Nipah virus Post-F protein is provided, the method comprising the following steps: culturing the transformant as described in the fourth aspect of the present invention under suitable conditions, and obtaining the antibody from the culture.

[0047] In the sixth aspect of the present invention, an immunoconjugate is provided, wherein the immunoconjugate comprises the antibody or antigen-binding fragment thereof as described in the first aspect of the present invention and a coupling portion.

[0048] In some embodiments, the conjugated moiety is selected from the group consisting of a detectable label, a drug, a cytokine, a radionuclide, an enzyme, a gold nanoparticle / nanorod, a nanomagnetic particle, a viral coat protein or a VLP, or a combination thereof.

[0049] In some embodiments, the radionuclide comprises a diagnostic isotope and / or a therapeutic isotope.

[0050] In some embodiments, the diagnostic isotope is selected from the group consisting of Tc-99m, Ga-68, F-18, I-123, I-125, I-131, In-111, Ga-67, Cu-64, Zr-89, C-11, Lu-177, Re-188, or a combination thereof; and / or, the therapeutic isotope is selected from the group consisting of Lu-177, Y-90, Ac-225, As-211, Bi-212, Bi-213, Cs-137, Cr-51, Co-60, Dy-165, Er-169, Fm-255, Au-198, Ho-166, 1-125, 1-131, Ir-192, Fe-59, Pb-212, Mo-99, Pd-103, P-32, K-42, Re-186, Re-188, Sm-153, Ra223, Ru-106, Na24, Sr89, Tb-149, Th-227, Xe-133, Yb-169, Yb-177, or a combination thereof.

[0051] In a seventh aspect of the present invention, a pharmaceutical composition is provided, comprising:

[0052] (i) the antibody or antigen-binding fragment thereof according to the first aspect of the present invention or the immunoconjugate according to the sixth aspect of the present invention; and

[0053] (ii) a pharmaceutically acceptable carrier.

[0054] In an eighth aspect of the present invention, a kit is provided, the kit comprising the antibody or antigen-binding fragment thereof as described in the first aspect of the present invention or the immunoconjugate as described in the sixth aspect of the present invention.

[0055] In the ninth aspect of the present invention, there is provided the use of the antibody or antigen-binding fragment thereof as described in the first aspect of the present invention, the isolated nucleic acid as described in the second aspect of the present invention, the recombinant expression vector as described in the third aspect of the present invention, the transformant as described in the fourth aspect of the present invention, the immunoconjugate as described in the sixth aspect of the present invention, the pharmaceutical composition as described in the seventh aspect of the present invention and / or the kit as described in the eighth aspect of the present invention for preparing (a) a reagent or kit for detecting Nipah virus; and / or (b) a drug for preventing and / or treating a disease or condition caused by Nipah virus infection.

[0056] In some embodiments, the reagent is a diagnostic reagent. In some embodiments, the diagnostic reagent is a detection sheet or a detection plate. In some embodiments, the diagnostic reagent is used to detect Nipah virus Post-F protein.

[0057] In the tenth aspect of the present invention, a method for treating a disease or condition caused by Nipah virus infection is provided, the method comprising administering to a subject in need thereof an effective amount of one or more of the antibody or antigen-binding fragment thereof as described in the first aspect of the present invention, the immunoconjugate as described in the sixth aspect of the present invention, the pharmaceutical composition as described in the seventh aspect of the present invention, and the kit as described in the eighth aspect of the present invention.

[0058] In some embodiments, the subject comprises a mammal, such as a human.

[0059] In some embodiments, the disease or condition caused by Nipah virus infection is selected from the group consisting of fever, severe headache, meningitis, or a combination thereof.

[0060] In the eleventh aspect of the present invention, there is provided an antibody or antigen-binding fragment thereof as described in the first aspect of the present invention, an isolated nucleic acid as described in the second aspect of the present invention, a recombinant expression vector as described in the third aspect of the present invention, a transformant as described in the fourth aspect of the present invention, an immunoconjugate as described in the sixth aspect of the present invention, a pharmaceutical composition as described in the seventh aspect of the present invention and / or a kit as described in the eighth aspect of the present invention for preventing and / or treating a disease or condition caused by Nipah virus infection in a subject.

[0061] In some embodiments, the subject comprises a mammal, such as a human.

[0062] In some embodiments, the disease or condition caused by Nipah virus infection is selected from the group consisting of fever, severe headache, meningitis, or a combination thereof.

[0063] In the twelfth aspect of the present invention, a method for detecting Nipah virus Post-F protein in a sample is provided, the method comprising: (i) contacting the sample with the antibody or antigen-binding fragment thereof as described in the first aspect of the present invention or the immunoconjugate as described in the sixth aspect of the present invention; and (ii) detecting whether an antigen-antibody complex is formed, wherein the formation of an antigen-antibody complex indicates the presence of Nipah virus Post-F protein in the sample.

[0064] In some embodiments, the detecting is an in vitro or ex vivo detecting.

[0065] In some embodiments, the detection is for non-diagnostic purposes.

[0066] In order to solve the above problems in the field, a technical solution adopted in a specific embodiment of the present invention is: using eukaryotic cells to express and purify post-F protein, and using it as an antigen to immunize mice. The mice are regularly bled, the spleen of the mouse with the best titer is taken, RNA is extracted, a phage immune library is constructed, and antibody screening is performed.

[0067] The antibody titer and immune effect were determined by indirect ELISA; the spleen of the mouse with the best titer was taken, the total RNA of the spleen cells was extracted and reverse transcribed into cDNA, which was used as a template to amplify the heavy and light chain variable region VH and VL gene fragments, and connected to Scfv through G4Slinker. After the Scfv gene fragment and the phage plasmid were connected, an anti-Scfv phage antibody library was constructed by electric shock, and panning was performed to enrich the phage with high affinity, and finally the anti-Post-F antibody was expressed and purified after recombination.

[0068] On the basis of being in accordance with the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain the preferred embodiments of the present invention.

[0069] The reagents and raw materials used in the present invention are commercially available.

[0070] The positive and progressive effect of the present invention is that the anti-Nipah virus specific monoclonal antibody of the present invention can specifically bind to the Post-F conformational protein and has the potential to be developed into a Nipah virus antibody detection kit or a virus neutralizing antibody drug. BRIEF DESCRIPTION OF THE DRAWINGS

[0071] Figure 1 This is a graph showing the mouse serum titer determination in Example 1 of the present application.

[0072] Figure 2 It is a non-reducing SDS-PAGE electrophoresis diagram of the monoclonal antibody of Example 2 of the present application.

[0073] Figure 3 It is the reduced SDS-PAGE electrophoresis diagram of the monoclonal antibody of Example 2 of the present application.

[0074] Figure 4 This is the antibody binding curve provided in Example 2 of the present application. DETAILED DESCRIPTION

[0075] In the present invention, unless otherwise specified, the scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. In addition, the molecular genetics, nucleic acid chemistry, chemistry, molecular biology, biochemistry, cell culture, microbiology, cell biology, genomics and recombinant DNA procedures used herein are conventional procedures widely used in the corresponding fields. At the same time, in order to better understand the present invention, the definitions and explanations of the relevant terms are provided below:

[0076] In one aspect of the present invention, an antibody or an antigen-binding fragment thereof that specifically targets the Nipah virus Post-F protein is provided.

[0077] In the present invention, the letters in the amino acid sequence represent the single-letter abbreviations of amino acids known in the art, such as those described in J.Biol.Chem, 243, p3558 (1968): alanine: Ala-A, arginine: Arg-R, aspartic acid: Asp-D, cysteine: Cys-C, glutamine: Gln-Q, glutamic acid: Glu-E, histidine: His-H, glycine: Gly-G, asparagine: Asn-N, tyrosine: Tyr-Y, proline: Pro-P, serine: Ser-S, methionine: Met-M, lysine: Lys-K, valine: Val-V, isoleucine: Ile-I, phenylalanine: Phe-F, leucine: Leu-L, tryptophan: Trp-W, threonine: Thr-T.

[0078] In the present invention, the amino acid sequences of complementarity determining regions (CDRs) listed are shown according to the definition of IMGT numbering rules. However, it is well known to those skilled in the art that the CDR of an antibody can be defined in the art by a variety of methods, such as based on the three-dimensional structure of the antibody and the topology of the CDR loop (Chothia et al. (1989) Nature 342:877-883); "Standard conformations for the canonical structures of immunoglobulins" published by Al-Lazikani et al. (Journal of Molecular Biology, 273, 927-948 (1997)); "Sequences of Proteins of Immunological Interest" published by Kabat et al. (USDepartment of Health and Human Services, National Institutes of Health (1987)); AbM (University of Bath), Contact (University College London), the international ImMunoGeneTics database (IMGT, imgt.cines.fr / on the World Wide Web); and the North CDR definition based on affinity propagation clustering using a large number of crystal structures. It will be appreciated by those skilled in the art that, unless otherwise specified, the terms "CDR" and "complementarity determining region" of a given antibody or region thereof (e.g., variable region) should be understood to encompass complementarity determining regions as defined by any of the above-mentioned known schemes described in the present invention.

[0079] 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). Although the scope of the present invention is based on the sequences shown in the definition according to the IMGT numbering rules, the amino acid sequences corresponding to the definition rules of other CDRs should also fall within the scope of the present invention.

[0080] In the present invention, the term "full-length antibody" is used interchangeably to refer to a glycoprotein comprising at least two heavy chains (HC) and two light chains (LC) interconnected by disulfide bonds. Each heavy chain consists of a heavy chain variable region (abbreviated as VH in the present invention) and a heavy chain constant region. The heavy chain constant region consists of three domains CH1, CH2 and CH3. Each light chain consists of a light chain variable region (abbreviated as VL in the present invention) and a light chain constant region (abbreviated as CL in the present invention). The light chain constant region consists of one domain CL. Mammalian heavy chains are classified as α, δ, ε, γ and μ. Mammalian light chains are classified as λ or κ. Immunoglobulins containing α, δ, ε, γ and μ heavy chains are classified as immunoglobulins IgA, IgD, IgE, IgG and IgM. Complete antibodies form a "Y" shape. The stem of the Y is composed of the second and third constant regions of the two heavy chains (and the fourth constant region for IgE and IgM) combined together, and a disulfide bond (interchain) is formed in the hinge. Heavy chains γ, α and δ have a constant region consisting of three tandem (in a row) Ig domains, and a hinge region for increased flexibility; heavy chains μ and ε have a constant region consisting of four immunoglobulin domains. The second and third constant regions are called "CH2 domains" and "CH3 domains", respectively. Each arm of the Y includes the variable region of a single heavy chain bound to the variable and constant regions of a single light chain and the first constant region. The variable regions of the light and heavy chains are responsible for antigen binding.

[0081] In the present invention, "Fab fragment" consists of one light chain and CH1 and variable region of one heavy chain. The heavy chain of the Fab molecule cannot form a disulfide bond with another heavy chain molecule. The "Fc" region contains two heavy chain fragments including the CH2 and CH3 domains of the antibody. The two heavy chain fragments are held together by two or more disulfide bonds and by the hydrophobic interaction of the CH3 domain. "Fab' fragment" contains one light chain and a portion of one heavy chain including the VH domain and the CH1 domain and the region between the CH1 and CH2 domains, thereby forming an interchain disulfide bond between the two heavy chains of the two Fab' fragments to form a F(ab')2 molecule. "F(ab')2 fragment" contains two light chains and two heavy chains including a portion of the constant region between the CH1 and CH2 domains, thereby forming an interchain disulfide bond between the two heavy chains. Therefore, the F(ab')2 fragment consists of two Fab' fragments held together by a disulfide bond between the two heavy chains. The term "Fv" means an antibody fragment consisting of the VL and VH domains of a single arm of an antibody, but lacking a constant region.

[0082] In the present invention, the scFv refers to a single chain antibody fragment, which includes a heavy chain variable region, a light chain variable region and a connecting peptide of 15 to 20 amino acids. The VL and VH domains are paired to form a monovalent molecule by a connecting peptide that enables them to be produced as a single polypeptide chain [see, for example, Bird et al., Science 242: 423-426 (1988) and Huston et al., Proc. Natl. Acad. Sci. USA 85: 5879-5883 (1988)]. Such scFv molecules may have a general structure: NH2-VL-connecting peptide-VH-COOH or NH2-VH-connecting peptide-VL-COOH.

[0083] In one aspect of the present invention, provided are nucleic acid molecules encoding the antibodies or antigen-binding fragments thereof of the present invention, vectors containing the nucleic acid molecules, and host cells and methods for expressing the antibodies or antigen-binding fragments thereof of the present invention.

[0084] In the present invention, "nucleic acid" refers to a nucleotide chain of any length, and includes DNA and RNA. The nucleotides can be deoxyribonucleotides, ribonucleotides, modified nucleotides or bases, and / or their analogs, or any substrate that can be incorporated into the chain by DNA or RNA polymerase.

[0085] In the present invention, the term "recombinant expression vector" means a genetically modified oligonucleotide or polynucleotide construct that permits expression of mRNA, protein, polypeptide or peptide by a host cell when the construct comprises a nucleotide sequence encoding mRNA, protein, polypeptide or peptide, and the vector contacts the cell under conditions sufficient to allow mRNA, protein, polypeptide or peptide to be expressed in the cell. The vector of the present invention is generally not naturally occurring. However, portions of the vector may be naturally occurring. The recombinant expression vector of the present invention may comprise any type of nucleotide, including but not limited to DNA and RNA, which may be single-stranded or double-stranded, synthesized or partially obtained from a natural source, and which may contain natural, non-natural or altered nucleotides. The recombinant expression vector may comprise naturally occurring or non-naturally occurring nucleotides, or both types of connections. In exemplary aspects, altered nucleotides or non-naturally occurring nucleotides are connected without hindering transcription or replication of the vector.

[0086] The recombinant expression vector of the present invention can be any suitable recombinant expression vector, which can be used for transformation or transfection to deliver one or more genes or sequences of interest into any suitable host cell and preferably express the gene or sequence in the host cell. Suitable vectors include those designed for expansion and amplification or for expression or both, examples of vectors include but are not limited to viral vectors, naked DNA or RNA expression vectors, plasmids, cosmids or phage vectors, DNA or RNA expression vectors associated with cationic coagulants, DNA or RNA expression vectors encapsulated in liposomes, and certain eukaryotic cells, such as production cells.

[0087] In the present invention, the term "host cell" refers to any type of cell that can contain the nucleic acid or vector described herein. In exemplary aspects, the host cell is a eukaryotic cell, such as a plant, an animal, a fungus, or an alga; or it can be a prokaryotic cell, such as a bacterium or a protozoan.

[0088] In another aspect of the present invention, a pharmaceutical composition is also provided.

[0089] In one embodiment, the pharmaceutical composition of the present invention comprises an antibody or an active fragment thereof or a fusion protein thereof of the present invention, and a pharmaceutically acceptable carrier. Typically, these substances can be formulated in a non-toxic, inert and pharmaceutically acceptable aqueous carrier medium, wherein the pH is generally about 5-8, preferably about 6-8, although the pH value may vary depending on the nature of the formulated substance and the condition to be treated. The formulated pharmaceutical composition can be administered by conventional routes, including (but not limited to): intraperitoneal, intravenous, or topical administration.

[0090] The pharmaceutical composition of the present invention contains a safe and effective amount (such as 0.001-99wt%, preferably 0.01-90wt%, more preferably 0.1-80wt%) of the above-mentioned antibody (or its conjugate) of the present invention and a pharmaceutically acceptable carrier or excipient.

[0091] Such carriers include (but are not limited to): saline, buffer, glucose, water, glycerol, ethanol, and combinations thereof. The pharmaceutical preparation should be compatible with the mode of administration. The pharmaceutical composition of the present invention can be prepared in the form of an injection, for example, by conventional methods using physiological saline or an aqueous solution containing glucose and other adjuvants. Pharmaceutical compositions such as injections and solutions are preferably manufactured under sterile conditions.

[0092] When using a pharmaceutical composition, a safe and effective amount of the antibody or antigen-binding fragment immunoconjugate of the present invention is administered to a mammal, wherein the safe and effective amount is usually at least about 10 μg / kg body weight, and in most cases does not exceed about 50 mg / kg body weight, preferably the dose is about 10 μg / kg body weight to about 10 mg / kg body weight. Of course, the specific dose should also take into account factors such as the route of administration and the patient's health status, which are all within the skill of a skilled physician.

[0093] In addition, the pharmaceutical composition of the present invention can also be used together with other therapeutic agents for treating diseases or conditions caused by Nipah virus infection.

[0094] The nipah postF protein is expressed and purified by eukaryotic cells, and used as an antigen to immunize mice. Blood is collected from mice regularly, and the spleen of the mouse with the best titer is taken. The total RNA of the spleen cells is extracted and reversely transcribed into cDNA, which is used as a template for amplification to obtain VH and VL gene fragments, which are constructed into scFv and connected with phage plasmids. An anti-nipah phage antibody library is constructed by electric shock, and panning, sequencing and expression purification are performed. The selected mouse antibodies are subjected to affinity screening, thereby obtaining the monoclonal antibody of the present invention. On this basis, the inventors have completed the present invention.

[0095] The present invention is further described below by way of examples, but the present invention is not limited to the scope of the examples. The experimental methods in the following examples without specifying specific conditions are carried out according to conventional methods and conditions, or selected according to the product specifications.

[0096] Reagents and instruments used in the examples:

[0097] Main reagents

[0098] PrimeScript 1st Strand cDNA Synthesis kit (6110A, takara), 5U / μl LA Taq (RR02MA, TaKaRa); TG1 Electrocompetent Cells4 (60502-2 DaKaRa); M-280 Streptavidin (11206D, Invitrogen); SfiI restriction endonuclease (R0123S, NEB), T4 DNA ligase (M0202L, NEB); M13KO7 helper phage (2N0315S Chengdu Renyu Biological); BSA (A600332-0100, Shanghai Shenggong Biotechnology Co., Ltd.); goat anti-mouse (C030237, Luoyang Bio-Tong Experimental Materials Center).

[0099] Main instruments

[0100] Microplate reader varioskan lux (source: Thermo); Nanodrop one (source: ThermoScientific); electroporator BLO-RAD (source: BIO-RAD); shaking incubator ZQZY-88AH (source: Shanghai Zhichu); high-speed centrifuge LYNX6000 (source: Thermo Scientific); biological clean bench BCM-1300A-Ⅱ (source: Suzhou Antai Air Technology Co., Ltd.).

[0101] The embodiments of the present application provide monoclonal antibodies to the postF protein, including seven monoclonal antibodies numbered #P3, #P4, #P5, #P6, #P7, #P9 and #P10.

[0102] The amino acid sequences of monoclonal antibodies #P3, #P4, #P5, #P6, #P7, #P9 and #P10 are shown in Tables 1-7. The following complementary determining region CDR amino acid sequences are shown in accordance with the definition of IMGT numbering rules.

[0103] Table 1 Amino acid sequence of antibody #P3

[0104]

[0105]

[0106] Table 2 Amino acid sequence of antibody #P4

[0107]

[0108] Table 3 Amino acid sequence of antibody #P5

[0109]

[0110]

[0111] Table 4 Amino acid sequence of antibody #P6

[0112]

[0113]

[0114] Table 5 Amino acid sequence of antibody #P7

[0115]

[0116] Table 6 Amino acid sequence of antibody #P9

[0117]

[0118]

[0119] Table 7 Amino acid sequence of antibody #P10

[0120]

[0121]

[0122] Table 8 Nucleotide sequence of antibody #P3

[0123]

[0124]

[0125] Table 9 Nucleotide sequence of antibody #P4

[0126]

[0127]

[0128]

[0129] Table 10 Nucleotide sequence of antibody #P5

[0130]

[0131]

[0132] Table 11 Nucleotide sequence of antibody #P6

[0133]

[0134]

[0135]

[0136] Table 12 Nucleotide sequence of antibody #P7

[0137]

[0138]

[0139] Table 13 Nucleotide sequence of antibody #P9

[0140]

[0141]

[0142]

[0143] Table 14 Nucleotide sequence of antibody #P10

[0144]

[0145]

[0146] Example 1: Preparation of anti-post-F protein monoclonal antibodies

[0147] The Nipah Post-F mouse monoclonal antibody in Table 1-7 of the present invention is prepared by the following steps:

[0148] 1.1 Selection of immunogens

[0149] The amino acid sequence verified by Nipah Post-F (RNAcure) is as follows (SEQ ID NO: 97):

[0150] MVVILDKRCYCNLLILILMISECSVGILHYEKLSKIGLVKGVTRKYKIKSNPLTKDIVIKMIPNVS NMSQCTGSVMENYKTRLNGILTPIKGALEIYKNNTHDLVGDVRATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVYVLTALQDYINTNLVPTIDKISC KQTELSLDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAISQAFGGNYETLLRTLGYATEDFDDLLESDSITGQIIYVDLSSYYIIVRVYFPILTEIQQAYIQELLPVSFNNDN SEWISIVPNFILVRNTLISNIEIGFCLITKRSVICNQDYATPMTNNMRECLTGSTEKCPRELVVSSHVPRFALSNGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTTCP TAVLGNVIISLGKYLGSVNYNSEGIAIGPPVFTDKVDISSQISSMNQSLQQSKDYIKEAQRLLDTVNPSLEVLFQGPHHHHHHSGSWSHPQFEKGGGSGGGSGGSAWSHPQFEK

[0151] 1.2 Mouse immunization and antibody acquisition

[0152] Niv Post-F was used as the specific antigen to prepare mouse monoclonal antibodies.

[0153] 1.2.1 Protein Immunization

[0154] Wipe the needle of the emulsifier twice with 75% alcohol (use a new cotton ball for the second time). The range of the single emulsified liquid volume is about 250μl to 20ml. Dilute the Nipah Post-F (PON-V52H3) protein to 2mg / mL, take 250μL protein and 250μL complete adjuvant or incomplete adjuvant, and use an emulsifier to emulsify, from top to bottom as a whole, and swing up and down locally. Generally, 1-10ml of emulsification volume can be emulsified in 3-5 minutes, and no more than 10 minutes.

[0155] Administration method: subcutaneous multiple-point immunization, and impact immunization is muscle immunization.

[0156] Strain: Balb / C, 6-8 weeks, female.

[0157] Quantity: 2 for control, 3 for sample, 2 for phage screening, 1 for backup.

[0158] Immunization dose: 100 μg / mouse / time, once every two weeks.

[0159] 1.2.2 Determination of mouse serum antibody titer

[0160] After the third immunization with Nipah Post-F, blood was collected one week apart to measure the mouse serum antibody titer. The specific method is as follows:

[0161] (1) Add 100uL of Nipah Post-F (PON-V52H3, Arco) antigen (100ng) into a 96-well plate for coating;

[0162] (2) Incubate overnight at 4°C in the dark;

[0163] (3) Wash the plate 4 times, adding 300uL 1×PBST each time. After the last wash, shake off as much as possible and blot on absorbent paper.

[0164] (4) Add 200 μL of 5% skim milk blocking solution to each well for blocking;

[0165] (5) Incubate at 37°C in the dark for 2 hours;

[0166] (6) Wash the plate four times, adding 300 μL 1× PBST each time. After the last wash, shake off as much as possible and blot on absorbent paper.

[0167] (7) Use the NAYO 96-well pipetting station to dilute the serum or cell supernatant into the blocked 96-well plate, 100 μL per well. For positive quality control screening, add 1000-fold diluted positive serum (without control antibody) to the cell supernatant.

[0168] (8) Incubate at 37°C in the dark for 1 h;

[0169] (9) Wash the plate 4 times, adding 300 μL 1× PBST each time. After the last wash, shake off as much as possible and blot on absorbent paper.

[0170] (10) Add 20,000-fold diluted goat anti-mouse IgG H+L secondary antibody to each well;

[0171] (11) Incubate at 37°C in the dark for 1 h;

[0172] (12) Wash the plate 4 times, adding 300 μL 1× PBST each time. After the last wash, shake off as much as possible and blot on absorbent paper.

[0173] (13) Add 100uL of TMB colorimetric solution (the colorimetric solution itself is colorless and cannot be used if it changes color);

[0174] (14) Incubate at 37°C in the dark for 10-15 min. Add 50 μL of stop solution to terminate the reaction and read the OD value at 450 nm.

[0175] like Figure 1 As shown, the serum of mice immunized with Nipah Pre-F mRNA-LNP was used as a negative control. The serum titers of mice No. 1-3 were all above 50,000, among which the serum titer of mouse No. 3 was the highest. Mice No. 1 and No. 3 were selected for phage screening, and the mouse sera were bound to Nipah Post-F (PON-V52H3) and Nipah Post-F (RNAcure) proteins.

[0176] 1.2.3 Immune library construction

[0177] Total RNA from mouse spleen cells was extracted and reverse transcribed into cDNA. The variable regions of heavy and light chains were amplified using cDNA as a template, connected to scfv with linkers and then connected to the phagemid vector. A 9-fold immune library was constructed by electroporation and packaged into scfv-phage for screening.

[0178] 1.2.4 Panning of phage library

[0179] After incubating scfv-phage with biotin-post F antigen at room temperature for 1 hour, the scfv-phage was added to the blocked SA-coated magnetic beads and incubated at room temperature for 15 minutes. The magnetic beads were washed 10 times with PBST, and then the specifically bound phages were eluted with trypsin at room temperature and used to infect E. coli TG1 in the logarithmic phase. After 3 rounds of panning, a highly enriched phage library was obtained.

[0180] 1.2.5 Screening of single positive phage

[0181] The selected phage enrichment library was used to pick out single clones one by one and packaged into scFv-phage in a 96-well round-bottom deep-well plate. Then, ELISA was performed with phage, and the positive wells were specific antibodies against Nipah Post-F antigen. The corresponding antibody sequence was obtained by sending the bacterial solution to the second-generation sequencing.

[0182] 1.2.6 Antibody clone construction and plasmid preparation

[0183] Primers were designed to amplify the heavy and light chain variable regions and loaded into a vector carrying the heavy and light chain constant regions of rabbit antibodies to construct a full antibody molecule, and a plasmid was prepared using a kit (MN740410.100).

[0184] Example 2: Antibody preparation and affinity determination

[0185] 2.1 Preparation of recombinant rabbit antibodies and determination of binding curves

[0186] The recombinant plasmid heavy and light chains were transfected into 293F cells at a ratio of 1:1. When the cell viability dropped to 80% on the fifth day, the supernatant was collected to purify the antibody.

[0187] Resuspend the magnetic beads by shaking or vortexing; take an appropriate amount of magnetic beads (L00965, GenScript magnetic beads loading 40mg / ml) and put them into a 50ml centrifuge tube, place it on the magnetic base of the magnetic stand, and discard the supernatant; add 10 times the volume of 0.1% PBST, invert and mix thoroughly, place it on the magnetic base of the magnetic stand, and discard the supernatant; repeat washing once, add an equal volume of PBST to resuspend for use; add the expression supernatant to the resuspended magnetic beads, rotate at 4℃ overnight; place it on the magnetic base of the magnetic stand, discard the supernatant or save it for use; wash three times with PBS, discard the supernatant Clear, and suck it clean with a 1ml pipette for the last time; add an equal volume of elution buffer, incubate at room temperature for 15 minutes, and resuspend 2-3 times during this period; place it on the magnetic base of the magnetic stand, collect the liquid into a clean 50ml centrifuge tube, and repeat the elution once; add neutralizing solution to the centrifuge tube at 100μl / ml, and mix gently; place the centrifuge tube on the magnetic base to adsorb the magnetic beads, use a 1ml gun to transfer the supernatant to the prepared EP tube, and measure the concentration; concentrate the antibody with an ultrafiltration tube, replace 1×PBS 3 times, suck out and measure the concentration, and store it in aliquots at -20℃.

[0188] 2.2 Antibody SDS-PAGE

[0189] Add water to the non-reduced and reduced samples to 9μl, add 3μL 4×LDS Buffer (NP0008, Thermo) to the non-reduced sample, and add 3μL 4×LDS Buffer (containing 10mM DTT) to the reduced sample. After mixing, boil the reduced sample at 95℃ for 10min. Take a piece of protein electrophoresis gel, tear off the protective film, insert the side with the switch, insert a baffle on the other side, turn off both switches at the same time, put it into the electrophoresis tank, add the newly prepared 1×MOPS buffer to the electrophoresis tank, and add 1×MOPS recovery solution to the outside. Unplug the comb, add 5ul protein marker to the first column, and then add the prepared sample to be tested. Note that the sample is added to the center of the well. Cover the lid, pay attention to the positive and negative poles, and stop the electrophoresis band at 170V55min when it runs below the red. Pour MOPS into the MOPS recovery liquid bottle, turn on the switch and take out the protein gel, pry it open with scissors, be careful to avoid the gel from drying, cut off the sample part with a comb, transfer the protein gel to clean water, boil, and repeat three times. Transfer the treated gel to Coomassie brilliant blue solution, boil, shake on a shaker for 5-30 minutes, change to clean water to de-brighten, until clear bands are visible.

[0190] Figure 2 , Figure 3 This is the SDS-PAGE electrophoresis diagram of the non-reduced and reduced forms of the purified Nipah Post-F antibodies (P3, P4, P5, P6, P7, P9 and P10) in this example. The antibodies have high purity and are further used for antibody performance measurement and analysis.

[0191] 2.3 Antibody binding curve determination

[0192] Prepare an antigen diluent with a final concentration of 1g / ml, mix well, and use a multichannel pipette to add 100μl per well to a 96-well plate, 100ng / well, and incubate overnight at 4℃. The next day, wash the plate three times with 0.05% PBST, then add 5% skim milk and block at 37℃ for 1-3 hours. After blocking, wash the plate three times with 0.05% PBST, add gradient diluted antibody samples, and incubate at 37℃ for 1 hour. After the primary antibody incubation is completed, wash the plate three times with 0.05% PBST, add 20,000-fold diluted goat anti-mouse IgG (H+L)-HRP, and incubate at 37℃ for 1 hour. After the secondary antibody incubation is completed, wash the plate three times with 0.05% PBST, add 50μL TMB, incubate at room temperature in the dark for 5-10 minutes, add 100μL 1M concentrated sulfuric acid to stop, and read the absorbance at 450nm.

[0193] Figure 4The binding curves of the Nipah Post-F antibodies (P3, P4, P5, P6, P7, P9 and P10) in this example and the control 13G5 (manufacturer: Absolute antibody, catalog number: Ab02857-1.1) were measured. As shown in the figure, the Nipah Post-F antibody binds strongly to the Post protein.

[0194] 2.4 Antibody affinity determination

[0195] According to the ELISA results, select the appropriate chip and use the manual mode to explore the binding Ru value of the antigen and antibody at a certain concentration. Use the anti-mouse antibody chip coupled with the Protein G chip (29179315, cytiva) or the CM5 chip (29104988, cytiva), select the Multi-cycle kinetics affinity using GST Capture kit program, set the injection and dissociation time, flow rate, regeneration solution, Startup cycle number, analyte concentration gradient and sample position. In Biacore Insight Evaluation Software, click Predefined to find the corresponding analysis method (Kinetics / affinity–Antibody / general–Multi-cycle kinetics affinity using GSTCapture kit) (corresponding to the experimental method), double-click or click the Open button in the lower right corner for analysis. The analysis software automatically performs data fitting, and the affinity of the screened antibodies with the purchased Nipah Post-F protein (PON-V52H3) and the homemade Nipah Post-F (RNAcure) are shown in Tables 8 and 9.

[0196] Table 8 Affinity determination of Nipah antibody and Nipah Post-F (PON-V52H3)

[0197]

[0198] Table 9 Affinity determination of Nipah antibody and Nipah Post-F (RNAcure)

[0199]

Claims

1. An antibody or antigen-binding fragment thereof against Nipah virus Post-F protein, characterized in that: The antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3; wherein the HCDR1, HCDR2 and HCDR3 respectively comprise the HCDR1, HCDR2 and HCDR3 in the heavy chain variable region whose amino acid sequence is any one of SEQ ID NO: 4, 13, 20, 25, 30, 34 or 42; and The LCDR1, LCDR2 and LCDR3 respectively comprise LCDR1, LCDR2 and LCDR3 in the light chain variable region of any one of SEQ ID NO: 9, 16, 23, 28, 32, 37 or 46.

2. The antibody or antigen-binding fragment thereof according to claim 1, wherein The HCDR1 comprises the amino acid sequence shown in SEQ ID NO: 1, 11, 18 or 39, the HCDR2 comprises the amino acid sequence shown in SEQ ID NO: 2, 12 or 40, the HCDR3 comprises the amino acid sequence shown in SEQ ID NO: 3, 19 or 41, the LCDR1 comprises the amino acid sequence shown in SEQ ID NO: 6 or 44, the LCDR2 comprises the amino acid sequence shown in SEQ ID NO: 7, and the LCDR3 comprises the amino acid sequence shown in SEQ ID NO: 8, 15, 22, 27, 36 or 45; Preferably, (i) the HCDR1 comprises the amino acid sequence shown in SEQ ID NO:1, the HCDR2 comprises the amino acid sequence shown in SEQ ID NO:2, the HCDR3 comprises the amino acid sequence shown in SEQ ID NO:3, the LCDR1 comprises the amino acid sequence shown in SEQ ID NO:6, the LCDR2 comprises the amino acid sequence shown in SEQ ID NO:7, and the LCDR3 comprises the amino acid sequence shown in SEQ ID NO:8; or (ii) the HCDR1 comprises the amino acid sequence shown in SEQ ID NO:11, the HCDR2 comprises the amino acid sequence shown in SEQ ID NO:12, the HCDR3 comprises the amino acid sequence shown in SEQ ID NO:3, the LCDR1 comprises the amino acid sequence shown in SEQ ID NO:6, the LCDR2 comprises the amino acid sequence shown in SEQ ID NO:7, and the LCDR3 comprises the amino acid sequence shown in SEQ ID NO:15; or (iii) the HCDR1 comprises the amino acid sequence shown in SEQ ID NO:18, the HCDR2 comprises the amino acid sequence shown in SEQ ID NO:2, the HCDR3 comprises the amino acid sequence shown in SEQ ID NO:19, the LCDR1 comprises the amino acid sequence shown in SEQ ID NO:6, the LCDR2 comprises the amino acid sequence shown in SEQ ID NO:7, and the LCDR3 comprises the amino acid sequence shown in SEQ ID NO:22; or (iv) the HCDR1 comprises the amino acid sequence shown in SEQ ID NO:11, the HCDR2 comprises the amino acid sequence shown in SEQ ID NO:12, the HCDR3 comprises the amino acid sequence shown in SEQ ID NO:3, the LCDR1 comprises the amino acid sequence shown in SEQ ID NO:6, the LCDR2 comprises the amino acid sequence shown in SEQ ID NO:7, and the LCDR3 comprises the amino acid sequence shown in SEQ ID NO:27; or (v) the HCDR1 comprises the amino acid sequence shown in SEQ ID NO:11, the HCDR2 comprises the amino acid sequence shown in SEQ ID NO:12, the HCDR3 comprises the amino acid sequence shown in SEQ ID NO:3, the LCDR1 comprises the amino acid sequence shown in SEQ ID NO:6, the LCDR2 comprises the amino acid sequence shown in SEQ ID NO:7, and the LCDR3 comprises the amino acid sequence shown in SEQ ID NO:15; or (vi) the HCDR1 comprises the amino acid sequence shown in SEQ ID NO:1, the HCDR2 comprises the amino acid sequence shown in SEQ ID NO:2, the HCDR3 comprises the amino acid sequence shown in SEQ ID NO:3, the LCDR1 comprises the amino acid sequence shown in SEQ ID NO:6, the LCDR2 comprises the amino acid sequence shown in SEQ ID NO:7, and the LCDR3 comprises the amino acid sequence shown in SEQ ID NO:36; or (vii) the HCDR1 comprises the amino acid sequence shown in SEQ ID NO:39, the HCDR2 comprises the amino acid sequence shown in SEQ ID NO:40, the HCDR3 comprises the amino acid sequence shown in SEQ ID NO:41, the LCDR1 comprises the amino acid sequence shown in SEQ ID NO:44, the LCDR2 comprises the amino acid sequence shown in SEQ ID NO:7, and the LCDR3 comprises the amino acid sequence shown in SEQ ID NO:

45.

3. The antibody or antigen-binding fragment thereof according to claim 1 or 2, characterized in that: The framework regions of the heavy chain variable region and / or light chain variable region are human, mouse or rabbit framework regions; Preferably, the heavy chain variable region comprises an amino acid sequence as shown in SEQ ID NO: 4, 13, 20, 25, 30, 34 or 42, or comprises an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with SEQ ID NO: 4, 13, 20, 25, 30, 34 or 42; and / or The light chain variable region comprises an amino acid sequence as shown in SEQ ID NO:9, 16, 23, 28, 32, 37 or 46, or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:9, 16, 23, 28, 32, 37 or 46; More preferably, (i) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO:4, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO:9; or (ii) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 13, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 16; or (iii) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 20, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 23; or (iv) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 25, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 28; or (v) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 30, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 32; or (vi) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 34, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 37; or (vii) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO:42, and the light chain variable region comprises 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, characterized in that The antibody or antigen-binding fragment thereof is a full-length antibody, Fab, Fab', F(ab')2 or Fv, and the Fv is preferably scFv; Preferably, the antibody is a full-length antibody; Further preferably, the amino acid sequence of the heavy chain of the antibody is shown in SEQ ID NO: 5, 14, 21, 26, 31, 35 or 43; and / or, the amino acid sequence of the light chain of the antibody is shown in SEQ ID NO: 10, 17, 24, 29, 33, 38 or 47; More preferably, (i) the amino acid sequence of the heavy chain of the antibody is shown in SEQ ID NO: 5, and the amino acid sequence of the light chain of the antibody is shown in SEQ ID NO: 10; or (ii) the amino acid sequence of the heavy chain of the antibody is shown in SEQ ID NO: 14, and the amino acid sequence of the light chain of the antibody is shown in SEQ ID NO: 17; or (iii) the amino acid sequence of the heavy chain of the antibody is shown in SEQ ID NO: 21, and the amino acid sequence of the light chain of the antibody is shown in SEQ ID NO: 24; or (iv) the amino acid sequence of the heavy chain of the antibody is shown in SEQ ID NO: 26, and the amino acid sequence of the light chain of the antibody is shown in SEQ ID NO: 29; or (v) the amino acid sequence of the heavy chain of the antibody is shown in SEQ ID NO:31, and the amino acid sequence of the light chain of the antibody is shown in SEQ ID NO:33; or (vi) the amino acid sequence of the heavy chain of the antibody is shown in SEQ ID NO: 35, and the amino acid sequence of the light chain of the antibody is shown in SEQ ID NO: 38; or (vii) the amino acid sequence of the heavy chain of the antibody is shown in SEQ ID NO:43, and the amino acid sequence of the light chain of the antibody is shown in SEQ ID NO:

47.

5. An isolated nucleic acid, characterized in that The isolated nucleic acid encodes the antibody or antigen-binding fragment thereof according to any one of claims 1 to 4; Preferably, the nucleotide sequence encoding the heavy chain of the antibody or its antigen-binding fragment is as shown in SEQ ID NO:52, 62, 69, 75, 80, 84 or 92; and / or, the nucleotide sequence encoding the light chain of the antibody or its antigen-binding fragment is as shown in SEQ ID NO:57, 65, 73, 78, 82, 87 or 96.

6. A recombinant expression vector, characterized in that: The recombinant expression vector comprises the isolated nucleic acid of claim 5; Preferably, the recombinant expression vector is a plasmid, a cosmid, a phage or a viral vector, and the viral vector is preferably a retroviral vector, a lentiviral vector, an adenoviral vector or an adeno-associated viral vector.

7. A transformant, characterized in that: The transformant comprises the isolated nucleic acid according to claim 5 or the recombinant expression vector according to claim 4, and the host cell of the transformant is a prokaryotic cell or a eukaryotic cell; Preferably, the host cell is a yeast cell or a mammalian cell, such as a HEK293 cell or a CHO cell; More preferably, the host cell is a 293F cell.

8. A method for preparing an antibody or an antigen-binding fragment thereof against Nipah virus Post-F protein, characterized in that: The method comprises the following steps: culturing the transformant according to claim 7 under appropriate conditions, and obtaining the antibody from the culture.

9. An immunoconjugate, characterized in that: The immunoconjugate comprises an antibody or antigen-binding fragment thereof as described in any one of claims 1 to 4 and a conjugated portion; Preferably, the coupling moiety is selected from the group consisting of a detectable marker, a drug, a cytokine, a radionuclide, an enzyme, a gold nanoparticle / nanorod, a nanomagnetic particle, a viral coat protein or a VLP, or a combination thereof; More preferably, the radionuclide comprises a diagnostic isotope and / or a therapeutic isotope; Further preferably, the diagnostic isotope is selected from the group consisting of Tc-99m, Ga-68, F-18, I-123, I-125, I-131, In-111, Ga-67, Cu-64, Zr-89, C-11, Lu-177, Re-188, or a combination thereof; and / or the therapeutic isotope is selected from the group consisting of Lu-177, Y-90, Ac-225, As-211, Bi-212, Bi-213, Cs-137, Cr-51, C o-60, Dy-165, Er-169, Fm-255, Au-198, Ho-166, 1-125, 1-131, Ir-192, Fe-59, Pb-212, Mo-99, Pd-103, P-32, K-42, Re-186, Re-188, Sm-153, Ra223, Ru-106, Na24, Sr89, Tb-149, Th-227, Xe-133, Yb-169, Yb-177, or a combination thereof.

10. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises: (i) the antibody or antigen-binding fragment thereof according to any one of claims 1 to 4 or the immunoconjugate according to claim 9; and (ii) a pharmaceutically acceptable carrier.

11. A kit, characterized in that: The kit comprises the antibody or antigen-binding fragment thereof according to any one of claims 1 to 4 or the immunoconjugate according to claim 9.

12. Use of the antibody or antigen-binding fragment thereof according to any one of claims 1 to 4, the isolated nucleic acid according to claim 5, the recombinant expression vector according to claim 6, the transformant according to claim 7, the immunoconjugate according to claim 9, the pharmaceutical composition according to claim 10 and / or the kit according to claim 11 for preparing (a) a reagent or kit for detecting Nipah virus; and / or (b) a drug for preventing and / or treating a disease or condition caused by Nipah virus infection; Preferably, the reagent is a diagnostic reagent; more preferably, the diagnostic reagent is a detection sheet or a detection plate; further more preferably, the diagnostic reagent is used to detect Nipah virus Post-F protein.

13. A method for detecting Nipah virus Post-F protein in a sample, characterized in that: The method comprises: (i) contacting the sample with the antibody or antigen-binding fragment thereof as described in any one of claims 1 to 4 or the immunoconjugate as described in claim 9; and (ii) detecting whether an antigen-antibody complex is formed, wherein the formation of the antigen-antibody complex indicates the presence of Nipah virus Post-F protein in the sample; Preferably, the detection is for non-diagnostic purposes.

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