Antibody specifically binding to rabies virus and application thereof

Abstract

The present disclosure provides antibodies or antigen-binding fragments thereof that specifically bind to rabies virus. The disclosure also relates to the use of said antibodies or antigen-binding fragments thereof in the treatment, prevention and / or diagnosis of rabies virus infection.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of biotechnology, and particularly relates to an antibody or antigen-binding fragment thereof specifically binding to rabies virus and application of the antibody or antigen-binding fragment thereof. BACKGROUND

[0002] Rabies is an acute central nervous system infectious disease caused by rabies virus (RABV), with a mortality rate close to 100%, and is considered as one of the most deadly infectious diseases. It is estimated that about 50,000 people die of rabies worldwide each year, of which about 40% are children under 15 years old. To date, rabies is still a serious public health challenge in more than 150 countries and regions worldwide, especially in Asia and Africa.

[0003] Rabies vaccines have achieved certain results in preventing the onset of the disease, but passive immunotherapy is still the most effective means for individuals who have been exposed to rabies virus. At present, the main drug for emergency prophylactic treatment of rabies is blood-derived immunoglobulin (RIG). However, the existing RIG treatment has many limitations, including high cost, supply shortage, potential pathogen contamination risk, and limited broad spectrum, which is difficult to cover the strains with far genetic distance in Lyssavirus.

[0004] Therefore, it is urgent to develop new humanized monoclonal antibodies against rabies virus G protein with broad-spectrum neutralizing activity, and to establish an efficient antibody discovery, screening and verification system to meet the needs of global rabies prevention and control. SUMMARY

[0005] The present disclosure provides an antibody specifically binding to rabies virus and application thereof. The antibody provided by the present application exhibits high affinity to G protein of various classic rabies virus vaccine strains, and has significant neutralizing ability to representative RABV strains and part of non-classic Lyssavirus strains in a pseudovirus system, with wide coverage and broad-spectrum neutralizing activity.

[0006] According to one aspect of the present disclosure, an antibody or antigen-binding fragment thereof specifically binding to rabies virus is provided, which comprises: (1) the following three heavy chain variable region complementarity determining regions (HCDRs): HCDR1, which has an amino acid sequence of HCDR1 contained in a heavy chain variable region as shown in any one of SEQ ID NOs: 1, 8, 15, 22, 29 and 36, or an amino acid sequence having substitution, deletion or addition of one or several amino acids compared with the amino acid sequence of HCDR1 contained in the heavy chain variable region; HCDR2 having an amino acid sequence of HCDR2 contained in a heavy chain variable region as shown in any one of SEQ ID NOs: 1, 8, 15, 22, 29, and 36, or an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared to the amino acid sequence of HCDR2 contained in the heavy chain variable region; HCDR3 having an amino acid sequence of HCDR3 contained in a heavy chain variable region as shown in any one of SEQ ID NOs: 1, 8, 15, 22, 29, and 36, or an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared to the amino acid sequence of HCDR3 contained in the heavy chain variable region; and (2) the following 3 light chain variable region complementarity determining regions (LCDRs): LCDR1 having an amino acid sequence of LCDR1 contained in a light chain variable region as shown in any one of SEQ ID NOs: 2, 9, 16, 23, 30, and 37, or an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared to the amino acid sequence of LCDR1 contained in the light chain variable region; LCDR2 having an amino acid sequence of LCDR2 contained in a light chain variable region as shown in any one of SEQ ID NOs: 2, 9, 16, 23, 30, and 37, or an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared to the amino acid sequence of LCDR2 contained in the light chain variable region; LCDR3 having an amino acid sequence of LCDR3 contained in a light chain variable region as shown in any one of SEQ ID NOs: 2, 9, 16, 23, 30, and 37, or an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared to the amino acid sequence of LCDR3 contained in the light chain variable region.

[0007] In some embodiments, the HCDR1-3 and / or the LCDR1-3 are defined by the rules of Kabat, AbM, Chothia, Contact, IMGT, or a combination thereof.

[0008] In some embodiments, the antibody or antigen-binding fragment thereof comprises: the 3 HCDRs contained in the heavy chain variable region set forth in SEQ ID NO: 1, or an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared to the amino acid sequence of the 3 HCDRs contained in the heavy chain variable region, or an amino acid sequence that has at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto, and the 3 LCDRs contained in the light chain variable region set forth in SEQ ID NO: 2, or an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared to the amino acid sequence of the 3 LCDRs contained in the light chain variable region, or an amino acid sequence that has at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto.

[0009] In some embodiments, the antibody or antigen-binding fragment thereof comprises: the 3 HCDRs contained in the heavy chain variable region set forth in SEQ ID NO: 8, or an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared to the amino acid sequence of the 3 HCDRs contained in the heavy chain variable region, or an amino acid sequence that has at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto, and the 3 LCDRs contained in the light chain variable region set forth in SEQ ID NO: 9, or an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared to the amino acid sequence of the 3 LCDRs contained in the light chain variable region, or an amino acid sequence that has at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto.

[0010] In some embodiments, the antibody or antigen-binding fragment thereof comprises: the 3 HCDRs contained in the heavy chain variable region set forth in SEQ ID NO: 15, or an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared to the amino acid sequence of the 3 HCDRs contained in the heavy chain variable region, or an amino acid sequence that has at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto, and the 3 LCDRs contained in the light chain variable region set forth in SEQ ID NO: 16, or an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared to the amino acid sequence of the 3 LCDRs contained in the light chain variable region, or an amino acid sequence that has at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto.

[0011] In some embodiments, the antibody or antigen-binding fragment thereof comprises: the 3 HCDRs contained in the heavy chain variable region of SEQ ID NO: 22, or an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared to the amino acid sequence of the 3 HCDRs contained in the heavy chain variable region, or an amino acid sequence having at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto, and the 3 LCDRs contained in the light chain variable region of SEQ ID NO: 23, or an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared to the amino acid sequence of the 3 LCDRs contained in the light chain variable region, or an amino acid sequence having at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto.

[0012] In some embodiments, the antibody or antigen-binding fragment thereof comprises: the 3 HCDRs contained in the heavy chain variable region of SEQ ID NO: 29, or an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared to the amino acid sequence of the 3 HCDRs contained in the heavy chain variable region, or an amino acid sequence having at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto, and the 3 LCDRs contained in the light chain variable region of SEQ ID NO: 30, or an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared to the amino acid sequence of the 3 LCDRs contained in the light chain variable region, or an amino acid sequence having at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto.

[0013] In some embodiments, the antibody or antigen-binding fragment thereof comprises: the 3 HCDRs contained in a heavy chain variable region set forth in SEQ ID NO: 36, or an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared to the amino acid sequence of the 3 HCDRs contained in said heavy chain variable region, or an amino acid sequence having at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto, and the 3 LCDRs contained in a light chain variable region set forth in SEQ ID NO: 37, or an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared to the amino acid sequence of the 3 LCDRs contained in said light chain variable region, or an amino acid sequence having at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto.

[0014] In some embodiments, the antibody or antigen-binding fragment thereof comprises: a HCDR1 having an amino acid sequence as set forth in any one of SEQ ID NOs: 3, 10, 17, 24, 31, or 38, an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared thereto, or an amino acid sequence having at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto; and / or, a HCDR2 having an amino acid sequence as set forth in any one of SEQ ID NOs: 4, 11, 18, 25, 32, or 39, an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared thereto, or an amino acid sequence having at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto; and / or, a HCDR3 having an amino acid sequence as set forth in any one of SEQ ID NOs: 5, 12, 19, 26, 33, or 40, an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared thereto, or an amino acid sequence having at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto; and / or, a LCDR1 having an amino acid sequence as set forth in any one of SEQ ID NOs: 6, 13, 20, 27, 34, or 41, an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared thereto, or an amino acid sequence having at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto; and / or, a LCDR2 having an amino acid sequence as set forth in WAS, ASS, GAS, EVT, EDN, or AAS, or an amino acid sequence having one amino acid substitution, deletion, or addition compared thereto; and / or, a LCDR3 having an amino acid sequence as set forth in any one of SEQ ID NOs: 7, 14, 21, 28, 35, or 42, an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared thereto, or an amino acid sequence having at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto.

[0015] In some embodiments, the antibody or antigen-binding fragment thereof comprises: HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, or an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared to the amino acid sequence of said HCDR1, HCDR2 and HCDR3, or an amino acid sequence having at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto, and / or LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO: 6, amino acid sequence WAS and SEQ ID NO: 7, or an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared to the amino acid sequence of said LCDR1, LCDR2 and LCDR3, or an amino acid sequence having at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto.

[0016] In some embodiments, the antibody or antigen-binding fragment thereof comprises: HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO: 10, SEQ ID NO: 11 and SEQ ID NO: 12, or an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared to the amino acid sequence of said HCDR1, HCDR2 and HCDR3, or an amino acid sequence having at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto, and / or LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO: 13, amino acid sequence ASS and SEQ ID NO: 14, or an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared to the amino acid sequence of said LCDR1, LCDR2 and LCDR3, or an amino acid sequence having at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto.

[0017] In some embodiments, the antibody or antigen-binding fragment thereof comprises: HCDR1, HCDR2, and HCDR3 as set forth in SEQ ID NO: 17, SEQ ID NO: 18, and SEQ ID NO: 19, or an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared to the amino acid sequence of the HCDR1, HCDR2, and HCDR3, or an amino acid sequence having at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto, and / or LCDR1, LCDR2, and LCDR3 as set forth in SEQ ID NO: 20, an amino acid sequence of GAS, and SEQ ID NO: 21, or an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared to the amino acid sequence of the LCDR1, LCDR2, and LCDR3, or an amino acid sequence having at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto.

[0018] In some embodiments, the antibody or antigen-binding fragment thereof comprises: HCDR1, HCDR2, and HCDR3 as set forth in SEQ ID NO: 24, SEQ ID NO: 25, and SEQ ID NO: 26, or an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared to the amino acid sequence of the HCDR1, HCDR2, and HCDR3, or an amino acid sequence having at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto, and / or LCDR1, LCDR2, and LCDR3 as set forth in SEQ ID NO: 27, an amino acid sequence of EVT, and SEQ ID NO: 28, or an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared to the amino acid sequence of the LCDR1, LCDR2, and LCDR3, or an amino acid sequence having at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto.

[0019] In some embodiments, the antibody or antigen-binding fragment thereof comprises: HCDR1, HCDR2, and HCDR3 as set forth in SEQ ID NO: 31, SEQ ID NO: 32, and SEQ ID NO: 33, or an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared to the amino acid sequence of the HCDR1, HCDR2, and HCDR3, or an amino acid sequence having at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto, and / or LCDR1, LCDR2, and LCDR3 as set forth in SEQ ID NO: 34, an amino acid sequence of EDN, and SEQ ID NO: 35, or an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared to the amino acid sequence of the LCDR1, LCDR2, and LCDR3, or an amino acid sequence having at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto.

[0020] In some embodiments, the antibody or antigen-binding fragment thereof comprises: HCDR1, HCDR2, and HCDR3 as set forth in SEQ ID NO: 38, SEQ ID NO: 39, and SEQ ID NO: 40, or an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared to the amino acid sequence of the HCDR1, HCDR2, and HCDR3, or an amino acid sequence having at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto, and / or LCDR1, LCDR2, and LCDR3 as set forth in SEQ ID NO: 41, an amino acid sequence of AAS, and SEQ ID NO: 42, or an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared to the amino acid sequence of the LCDR1, LCDR2, and LCDR3, or an amino acid sequence having at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto.

[0021] In some embodiments, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 1, an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared thereto, or an amino acid sequence having at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto; and / or, a light chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 2, an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared thereto, or an amino acid sequence having at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto.

[0022] In some embodiments, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 1, an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared thereto, or an amino acid sequence having at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto; and a light chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 2, an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared thereto, or an amino acid sequence having at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto.

[0023] In some embodiments, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 8, an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared thereto, or an amino acid sequence having at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto; and a light chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 9, an amino acid sequence having one or several amino acid substitutions, deletions, or additions compared thereto, or an amino acid sequence having at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto.

[0024] In some embodiments, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 15, an amino acid sequence that differs from that by one or several amino acid substitutions, deletions, or additions, or an amino acid sequence that has at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto; and a light chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 16, an amino acid sequence that differs from that by one or several amino acid substitutions, deletions, or additions, or an amino acid sequence that has at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto.

[0025] In some embodiments, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 22, an amino acid sequence that differs from that by one or several amino acid substitutions, deletions, or additions, or an amino acid sequence that has at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto; and a light chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 23, an amino acid sequence that differs from that by one or several amino acid substitutions, deletions, or additions, or an amino acid sequence that has at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto.

[0026] In some embodiments, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 29, an amino acid sequence that differs from that by one or several amino acid substitutions, deletions, or additions, or an amino acid sequence that has at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto; and a light chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 30, an amino acid sequence that differs from that by one or several amino acid substitutions, deletions, or additions, or an amino acid sequence that has at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto.

[0027] In some embodiments, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 36, an amino acid sequence that has one or several amino acid substitutions, deletions, or additions compared thereto, or an amino acid sequence that has at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto; and a light chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 37, an amino acid sequence that has one or several amino acid substitutions, deletions, or additions compared thereto, or an amino acid sequence that has at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, 100% sequence identity thereto.

[0028] In some embodiments, the antibody is of the IgA, IgD, IgE, IgG, or IgM type.

[0029] In some embodiments, the antibody or antigen-binding fragment thereof is a scFv, Fab, Fab', (Fab')2, Fv fragment, dsFv, diabody, bispecific antibody, and multispecific antibody.

[0030] In some embodiments, the antibody or antigen-binding fragment thereof is a chimeric antibody, human antibody, or humanized antibody.

[0031] In some embodiments, the antibody or antigen-binding fragment thereof further comprises a heavy chain constant region and / or a light chain constant region.

[0032] In some embodiments, the heavy chain constant region is selected from an IgGl, IgG2, IgG3, or IgG4 heavy chain constant region.

[0033] In some embodiments, the heavy chain constant region is selected from an hIgGl, hIgG2, hIgG3, or hIgG4 heavy chain constant region.

[0034] In some embodiments, the antibody or antigen-binding fragment thereof is capable of binding to Lyssavirus. In some embodiments, the antibody or antigen-binding fragment thereof is capable of binding to a RABV strain and / or a non-classical Lyssavirus strain. In some embodiments, the antibody or antigen-binding fragment thereof is capable of binding to a G protein of a Lyssavirus RABV strain.

[0035] In some embodiments, the antibody or antigen-binding fragment thereof has strong neutralization ability to G protein of RABV strains (e.g., P08667.1, CAI43218.1, ACR39382.1, ADM32132.1, ACR39382.1, ADJ29911.1, ADD84785.1, CAI43218.1, P08667.1, etc.), G protein of non-classical Lyssavirus strains (ABLV: KU739052.1, DUVV: KU761302.1, EBLV1: EU352768.1, EBLV2: AAX62813.1, IRKV: EF614260.1, MOKV: NC006429.1, etc.).

[0036] According to yet another aspect of the present disclosure, there is provided an antibody composition comprising a first antibody and a second antibody, the first antibody and / or the second antibody being selected from the antibodies or antigen-binding fragments thereof described in the present disclosure.

[0037] In some embodiments, the first antibody comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 10, SEQ ID NO: 11 and SEQ ID NO: 12, and LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 13, amino acid sequence of ASS and SEQ ID NO: 14; and the second antibody comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 31, SEQ ID NO: 32 and SEQ ID NO: 33, and LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 34, amino acid sequence of EDN and SEQ ID NO: 35.

[0038] In some embodiments, the first antibody comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 17, SEQ ID NO: 18 and SEQ ID NO: 19, and LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 20, amino acid sequence of GAS and SEQ ID NO: 21; and the second antibody comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 6, amino acid sequence of WAS and SEQ ID NO: 7.

[0039] In some embodiments, the first antibody comprises HCDR1, HCDR2, and HCDR3 as set forth in SEQ ID NO: 38, SEQ ID NO: 39, and SEQ ID NO: 40, and LCDR1, LCDR2, and LCDR3 as set forth in SEQ ID NO: 41, the amino acid sequence of AAS, and SEQ ID NO: 42; and the second antibody comprises HCDR1, HCDR2, and HCDR3 as set forth in SEQ ID NO: 3, SEQ ID NO: 4, and SEQ ID NO: 5, and LCDR1, LCDR2, and LCDR3 as set forth in SEQ ID NO: 6, the amino acid sequence of WAS, and SEQ ID NO: 7.

[0040] In some embodiments, the first antibody comprises HCDR1, HCDR2, and HCDR3 as set forth in SEQ ID NO: 17, SEQ ID NO: 18, and SEQ ID NO: 19, and LCDR1, LCDR2, and LCDR3 as set forth in SEQ ID NO: 20, the amino acid sequence of GAS, and SEQ ID NO: 21; and the second antibody comprises HCDR1, HCDR2, and HCDR3 as set forth in SEQ ID NO: 24, SEQ ID NO: 25, and SEQ ID NO: 26, and LCDR1, LCDR2, and LCDR3 as set forth in SEQ ID NO: 27, the amino acid sequence of EVT, and SEQ ID NO: 28.

[0041] In some embodiments, the first antibody comprises HCDR1, HCDR2, and HCDR3 as set forth in SEQ ID NO: 38, SEQ ID NO: 39, and SEQ ID NO: 40, and LCDR1, LCDR2, and LCDR3 as set forth in SEQ ID NO: 41, the amino acid sequence of AAS, and SEQ ID NO: 42; and the second antibody comprises HCDR1, HCDR2, and HCDR3 as set forth in SEQ ID NO: 24, SEQ ID NO: 25, and SEQ ID NO: 26, and LCDR1, LCDR2, and LCDR3 as set forth in SEQ ID NO: 27, the amino acid sequence of EVT, and SEQ ID NO: 28.

[0042] In some embodiments, the first antibody comprises a HCDR1, HCDR2, and HCDR3 as set forth in SEQ ID NO: 17, SEQ ID NO: 18, and SEQ ID NO: 19, and a LCDR1, LCDR2, and LCDR3 as set forth in SEQ ID NO: 20, the amino acid sequence of GAS, and SEQ ID NO: 21; and the second antibody comprises a HCDR1, HCDR2, and HCDR3 as set forth in SEQ ID NO: 31, SEQ ID NO: 32, and SEQ ID NO: 33, and a LCDR1, LCDR2, and LCDR3 as set forth in SEQ ID NO: 34, the amino acid sequence of EDN, and SEQ ID NO: 35.

[0043] In some embodiments, the first antibody comprises a heavy chain variable region as set forth in SEQ ID NO: 8 and a light chain variable region as set forth in SEQ ID NO: 9, and the second antibody comprises a heavy chain variable region as set forth in SEQ ID NO: 29 and a light chain variable region as set forth in SEQ ID NO: 30.

[0044] In some embodiments, the first antibody comprises a heavy chain variable region as set forth in SEQ ID NO: 15 and a light chain variable region as set forth in SEQ ID NO: 16, and the second antibody comprises a heavy chain variable region as set forth in SEQ ID NO: 1 and a light chain variable region as set forth in SEQ ID NO: 2.

[0045] In some embodiments, the first antibody comprises a heavy chain variable region as set forth in SEQ ID NO: 36 and a light chain variable region as set forth in SEQ ID NO: 37, and the second antibody comprises a heavy chain variable region as set forth in SEQ ID NO: 1 and a light chain variable region as set forth in SEQ ID NO: 2.

[0046] In some embodiments, the first antibody comprises a heavy chain variable region as set forth in SEQ ID NO: 15 and a light chain variable region as set forth in SEQ ID NO: 16, and the second antibody comprises a heavy chain variable region as set forth in SEQ ID NO: 22 and a light chain variable region as set forth in SEQ ID NO: 23.

[0047] In some embodiments, the first antibody comprises a heavy chain variable region as set forth in SEQ ID NO: 36 and a light chain variable region as set forth in SEQ ID NO: 37, and the second antibody comprises a heavy chain variable region as set forth in SEQ ID NO: 22 and a light chain variable region as set forth in SEQ ID NO: 23.

[0048] In some embodiments, the first antibody comprises a heavy chain variable region as set forth in SEQ ID NO: 15 and a light chain variable region as set forth in SEQ ID NO: 16, and the second antibody comprises a heavy chain variable region as set forth in SEQ ID NO: 29 and a light chain variable region as set forth in SEQ ID NO: 30.

[0049] According to another aspect of the present disclosure, there is provided a protein comprising an antigen binding domain, the antigen binding domain comprising an antibody or antigen binding fragment thereof as described in the present disclosure or an antibody composition as described.

[0050] In some embodiments, the protein is a chimeric antigen receptor comprising an antigen binding domain, a transmembrane domain, and an intracellular signaling domain, the antigen binding domain comprising an antibody or antigen binding fragment thereof as described in the present disclosure or an antibody composition as described.

[0051] In some embodiments, the protein is a multispecific antibody or antigen binding fragment thereof comprising two or more (e.g., three or four) antigen binding domains, wherein one antigen binding domain comprises an antibody or antigen binding fragment thereof as described in the present disclosure or an antibody composition as described.

[0052] According to yet another aspect of the present disclosure, there is provided a biological material related to an antibody or antigen binding fragment thereof as described in the present disclosure, an antibody composition as described, a chimeric antigen receptor as described, or a multispecific antibody or antigen binding fragment thereof as described, the biological material comprising any one of a1) - a9): a1) a nucleic acid molecule encoding an antibody or antigen binding fragment thereof as described in the present disclosure, an antibody composition as described, or a protein as described; a2) an expression cassette comprising the nucleic acid molecule of a1); a3) a vector comprising the nucleic acid molecule of a1); a4) a vector comprising the expression cassette of a2); a5) a cell comprising the nucleic acid molecule of a1); a6) a cell comprising the expression cassette of a2); a7) a cell comprising the vector of a3); a8) a cell comprising the vector of a4); a9) a cell comprising an antibody or antigen binding fragment thereof as described in the present disclosure, an antibody composition as described, or a protein as described; Any of the cells of a5) - a9) do not comprise reproductive material.

[0053] In some embodiments, the vector can be a eukaryotic cell vector and / or a prokaryotic cell vector, such as a retroviral vector, a lentiviral vector, a bacteriophage vector, an adenoviral vector, an adeno-associated vector, or a herpes simplex vector.

[0054] In some embodiments, the vector is present in a nanoparticle, a liposome, an exosome, a microvesicle, or a gene gun.

[0055] In some embodiments, the cell is a host cell that is conventional in the art, as long as the expression vector can stably express the nucleic acid molecule carried by the expression vector as the antibody or antigen-binding fragment thereof of the present application. Preferably, the host cell is a prokaryotic cell, preferably an E. coli cell such as TG1, BL21 (for expressing single-chain antibody or Fab antibody), or a eukaryotic cell, preferably a HEK293 cell or a CHO cell (for expressing full-length IgG antibody). The host cell of the present application can be obtained by transforming the expression vector into the host cell. The transformation method is conventional in the art, and preferably is chemical transformation, heat shock, or electroporation.

[0056] According to yet another aspect of the present disclosure, there is provided an antibody drug conjugate comprising: the antibody or antigen-binding fragment thereof of the present disclosure, or the antibody composition, and a drug covalently linked to the antibody or antigen-binding fragment thereof.

[0057] According to yet another aspect of the present disclosure, there is provided a pharmaceutical composition comprising: the antibody or antigen-binding fragment thereof of the present disclosure, the antibody composition, the chimeric antigen receptor, the multispecific antibody or antigen-binding fragment thereof, the biomaterial, or the antibody drug conjugate; and a pharmaceutically acceptable carrier.

[0058] In some embodiments, the pharmaceutically acceptable carrier can be conventional in the art, and can be any suitable physiologically or pharmaceutically acceptable pharmaceutical excipient. The pharmaceutical excipient is conventional in the art, and preferably includes a pharmaceutically acceptable excipient, a filler, or a diluent, etc. More preferably, the pharmaceutical composition comprises 0.01-99.99% of the antibody or antigen-binding fragment thereof and / or other small molecule drugs or antibodies or polypeptides, and 0.01-99.99% of a pharmaceutical carrier, the percentages being mass percentages of the pharmaceutical composition.

[0059] In some embodiments, the pharmaceutical composition can be administered parenterally, by injection or orally. The pharmaceutical composition can be prepared in a form suitable for administration, such as a solid, semi-solid or liquid form, and can be in the form of an aqueous solution, non-aqueous solution or suspension, a powder, tablet, capsule, granule, injection or infusion. It can be administered intravascularly, subcutaneously, intraperitoneally, intramuscularly, by inhalation, intranasally, airway instillation or intrathoracically. The pharmaceutical composition can also be administered in the form of an aerosol or spray, such as intranasally; or intrathecally, intramedullary or intraventricularly. It can also be administered transdermally, transcutaneously, topically, enterally, intravaginally, sublingually or rectally. The pharmaceutical composition can be prepared in various dosage forms as needed, and the dosage that is beneficial to the patient can be determined by a physician according to the patient's species, age, weight and general disease condition, administration mode and other factors.

[0060] In some embodiments, the antibody or antigen-binding fragment thereof in the pharmaceutical composition can be administered simultaneously or sequentially with other active ingredients.

[0061] According to yet another aspect of the present disclosure, a diagnostic or therapeutic kit is provided, comprising: an antibody or antigen-binding fragment thereof of the present disclosure, an antibody composition of the present disclosure, a chimeric antigen receptor of the present disclosure, a multispecific antibody or antigen-binding fragment thereof of the present disclosure, a biomaterial of the present disclosure, an antibody drug conjugate of the present disclosure, or a pharmaceutical composition of the present disclosure.

[0062] In some embodiments, the kit comprises one or more antibodies or antigen-binding fragments thereof of the present disclosure.

[0063] According to yet another aspect of the present disclosure, the use of an antibody or antigen-binding fragment thereof of the present disclosure, an antibody composition of the present disclosure, a chimeric antigen receptor of the present disclosure, a multispecific antibody or antigen-binding fragment thereof of the present disclosure, a biomaterial of the present disclosure, an antibody drug conjugate of the present disclosure, or a pharmaceutical composition of the present disclosure in the preparation of a product for any one of b1) - b3): b1) preventing, treating and / or diagnosing a rabies virus infection, b2) diagnosing a rabies virus infection or a disease caused thereby; b3) detecting the presence or level of a rabies virus in a sample.

[0064] In some embodiments, the rabies virus infection comprises rabies.

[0065] In some embodiments, the sample is whole blood, red blood cell concentrate, platelet concentrate, white blood cell concentrate, tissue, bone marrow aspirate, plasma, serum, cerebrospinal fluid, feces, urine, cultured cells, saliva, oral secretion and / or nasal secretion.

[0066] According to yet another aspect of the present disclosure, a method for detecting a Lyssavirus in a sample is provided, comprising the step of detecting the Lyssavirus in the sample with the antibody or antigen-binding fragment thereof of the first aspect.

[0067] In some embodiments, the sample is whole blood, red blood cell concentrate, platelet concentrate, white blood cell concentrate, tissue, bone marrow aspirate, plasma, serum, cerebrospinal fluid, stool, urine, cultured cells, saliva, oral secretion, and / or nasal secretion.

[0068] According to yet another aspect of the present disclosure, a method for preventing, treating, and / or diagnosing a Lyssavirus infection is provided, comprising administering to a subject an effective amount of the antibody or antigen-binding fragment thereof of the present disclosure, the antibody composition, the chimeric antigen receptor, the multispecific antibody or antigen-binding fragment thereof, the biomaterial, the antibody drug conjugate, or the pharmaceutical composition of the present disclosure.

[0069] In some embodiments, the Lyssavirus infection comprises rabies. BRIEF DESCRIPTION OF DRAWINGS

[0070] Figure 1 Monoclonal antibody RABV pseudovirus neutralization results are shown.

[0071] Figure 2 Monoclonal antibody ELISA binding results are shown.

[0072] Figure 3 Monoclonal antibody RABV pseudovirus neutralization results are shown.

[0073] Figure 4 Monoclonal antibody BLI evaluation results are shown.

[0074] Figure 5 Monoclonal antibody RABV street pseudovirus neutralization results are shown.

[0075] Figure 6 Monoclonal antibody Lyssavirus other virus pseudovirus neutralization results are shown. DETAILED DESCRIPTION

[0076] To obtain specific antibodies against the G protein of rabies virus, the present disclosure utilizes flow cytometry to screen G protein-specific memory B cells from the PBMC of vaccinees with different immune backgrounds. The selected individual B cells are sorted into 96-well PCR plates, subjected to single-cell lysis and RT-PCR reaction, and successfully construct cDNA containing naturally paired heavy and light chain variable region sequences. Subsequently, the full-length sequences of the antibody heavy and light chains are obtained by specific primer amplification, and an expression frame (TAD) is constructed by fragment splicing. The resulting product is transiently transfected into HEK293T cells, the antibody is expressed in a 96-well plate on a small scale, and the cell supernatant is collected for pseudovirus neutralization experiments for functional primary screening. The neutralization experiment uses a rabies virus VSV pseudovirus system to evaluate the neutralization ability of the antibody against representative rabies virus strains. Some of the expression products show good neutralization activity, thereby preliminarily screening a batch of positive candidate antibodies Figure 1 ).

[0077] To further screen candidate antibodies with strong binding ability and neutralization activity, ELISA binding experiments and pseudovirus neutralization experiments were performed on the preliminarily screened positive antibodies. ELISA results show that most antibodies have good binding ability to the G protein of the classical vaccine strain (P08667.1, CAI43218.1, ACR39382.1), and some antibodies exhibit broad-spectrum binding ability between different strains Figure 2 ).

[0078] Pseudovirus neutralization experiments further evaluated the neutralization activity of candidate antibodies against the G protein of multiple rabies virus strains (ADM32132.1, ACR39382.1, ADJ29911.1, ADD84785.1, CAI43218.1, P08667.1), and the results showed that some antibodies had strong neutralization ability against multiple strains (such as #230, #272, #249, #296, #213, #253), suggesting their potential as broad-spectrum neutralizing antibodies Figure 3 ).

[0079] To determine the affinity of purified antibodies to the G protein of rabies virus, a biological layer interference (BLI) experiment was performed. The results showed that most antibodies exhibited high affinity to the G protein of the vaccine strain (P08667.1) Figure 4 To verify the broad-spectrum neutralization ability of candidate antibodies in a real infection scenario, a pseudovirus neutralization screening system containing 46 street strains of RABV was constructed. The experimental results showed that most candidate antibodies retained neutralization ability in the street strain background, and some antibody combinations (such as #230 / #272, #249 / #213, #296 / #213, #249 / #253, #296 / #253, #249 / #272) exhibited synergistic or complementary neutralization effects between strains Figure 5). These results suggest that the antibody combination strategy can further expand the protection coverage and is a key direction for the optimization of rabies virus biological agents.

[0080] In addition, to determine whether these antibodies have neutralizing effects on other viruses of the Lyssavirus genus known to infect humans, the antibodies were further tested for neutralizing activity against the G protein of various Lyssavirus strains (ABLV: KU739052.1, ARAV: EF614259.1, BBLV: KU761304.1, EBLV1: EU352768.1, EBLV2: AAX62813.1, IRKV: EF614260.1, KHUV: EF614261.1). The results showed that some antibodies maintained high neutralizing potency against various Lyssavirus strains, and some combinations may exhibit potential broad-spectrum neutralization advantages. Figure 6 ).

[0081] The humanized antibodies obtained by screening of the present disclosure exhibit high affinity for the G protein of various classical vaccine strains in ELISA binding experiments, and have significant neutralizing ability for representative RABV strains and some non-classical Lyssavirus strains in a pseudovirus system, with a wide coverage range and potential as broad-spectrum neutralizing antibodies.

[0082] In the pseudovirus neutralization screening system of 46 naturally derived RABV street viruses, the protective ability of the candidate antibodies and their combinations was further verified in a context closer to natural infection, and some antibody combinations exhibited synergistic or complementary neutralization effects among different strains, significantly expanding the neutralization coverage. Moreover, the antibodies obtained by the present disclosure are all fully humanized IgG1 frameworks, suitable for large-scale mammalian cell expression, with low safety and immunogenicity risks, mature production processes, and can be used as candidate clinical prevention or treatment preparations to enter the drug development process.

[0083] To make the objectives, technical solutions and advantages of the present application clearer, further detailed description will be made in combination with the drawings and examples. The specific examples described herein are only used to explain the present application and do not constitute any limitation on the present application. In addition, in the following description, the description of well-known structures and technologies is omitted to avoid unnecessary confusion of the concept of the present disclosure. Such structures and technologies are also described in many publications.

[0084] Unless otherwise defined, all technical and scientific terms used in the present application have the same meaning as commonly used in the art to which the present application belongs. For the purpose of interpreting the present specification, the following definitions will apply, and terms used in the singular will also include the plural, and vice versa, if appropriate.

[0085] As used herein, the terms "a" and "one" include plural references unless the context clearly dictates otherwise. For example, reference to "a cell" includes a plurality of such cells, and equivalents thereof known to those skilled in the art, and so on.

[0086] As used herein, the term "about" means ±20% of the number that it precedes. In some embodiments, the term "about" means ±10% of the number that it precedes. In some embodiments, the term "about" means ±5% of the number that it precedes.

[0087] As used herein, the term "rabies virus" refers to the genus Lyssavirus. The genus Lyssavirus is a genus of viruses under the family Rhabdoviridae, whose member viruses can infect mammals, with the core feature of being highly pathogenic.

[0088] Rabies Virus (RABV) is the most classic, most pathogenic to humans, and most widely distributed virus species in the genus Lyssavirus, and is also the main pathogen of human rabies worldwide. Non-classical Lyssavirus is other virus species in the genus Lyssavirus except RABV.

[0089] Rabies Virus (RABV) belongs to the genus Lyssavirus of the family Rhabdoviridae, and is the pathogen of the zoonotic and highly infectious disease "rabies". It is mainly transmitted through the saliva of sick animals (such as dogs, cats, and bats), and the mortality rate after infection is almost 100%. The virus particle structure is composed of a nucleocapsid and an envelope. The nucleocapsid contains a single-stranded negative-strand RNA genome, which encodes five proteins: N, P, M, G, and L. The envelope surface is inlaid with the key glycoprotein G, which is the "key" for the virus to invade host cells and is also the core antigen for inducing the production of neutralizing antibodies in the body.

[0090] Street Virus refers to wild-type RABV isolated from naturally infected hosts (such as the brain tissue and saliva of sick animals), which is opposite to "fixed virus" (adapted to cell culture, with reduced pathogenicity, used for vaccine production) that has been long-term passaged and domesticated in the laboratory. Street virus has natural pathogenicity, strong virulence, and a wide host range (can infect dogs, cats, humans, bats, and other mammals), and the genome may have natural variation, which is more characteristic of the real epidemic virus strain.

[0091] Pseudovirus, also known as "pseudo virus", is an artificially constructed, non-replicating virus simulation particle, the core feature is "infectious but non-pathogenic". Structurally, only the "infection-related structure" of the virus (such as the envelope glycoprotein G of RABV) is retained, and the "replication essential genomic fragment" (such as the RNA genome of RABV is replaced by a reporter gene, or the coding sequence of L, P and other replication key proteins is deleted) is replaced or deleted. Functionally: can combine host cell receptors and invade cells through surface antigens (such as G protein) like natural viruses, but due to the lack of complete genome, it cannot complete replication in cells, nor can it form new infectious particles, and it will not cause diseases.

[0092] The term "antibody" as used herein encompasses a variety of antibody structures, including, but not limited to, monoclonal antibodies, polyclonal antibodies, monospecific and multispecific antibodies (e.g., bispecific or trispecific antibodies), single chain molecules, and antibody fragments, as long as they exhibit the desired antigen-binding activity.

[0093] The term "monoclonal antibody" as used herein refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical and / or bind the same epitope, except for possible variants containing naturally occurring mutations or arising during production of the monoclonal antibody preparation, typically present at low frequency. Unlike polyclonal antibody preparations, which typically include different antibodies directed against different antigenic determinants (epitopes), each monoclonal antibody of a monoclonal antibody preparation is directed against a single determinant on the antigen.

[0094] The terms "antibody or antigen-binding fragment thereof" and "antibody" are used interchangeably herein to refer to antibodies that are substantially similar to native antibodies. A "native antibody" refers to an immunoglobulin molecule that naturally occurs. For example, a native IgG class antibody is a heterotetrameric glycoprotein of about 150,000 daltons, composed of two light chains and two heavy chains that are disulfide-bonded. From N- to C-terminus, each heavy chain has a variable region (VH), also called a variable heavy chain domain or a heavy chain variable domain, and three constant domains (CH1, CH2 and CH3), also called a heavy chain constant region. From N- to C-terminus, each light chain has a variable region (VL), also called a variable light chain domain or a light chain variable domain, and a light chain constant domain (CL), also called a light chain constant region. The heavy chains of an antibody can be one of five types, a (IgA), d (IgD), e (IgE), g (IgG), or m (IgM), and can be further divided into subtypes, e.g., g1 (IgG1), g2 (IgG2), g3 (IgG3), g4 (IgG4), a1 (IgA1), and a2 (IgA2). The light chain of an antibody can be one of two types, a k light chain and a l light chain, based on the amino acid sequence of its constant domain.

[0095] Within the light and heavy chains, the variable and constant regions are joined by a "J" region of about 12 or more amino acids, with the heavy chain also including a "D" region of about 3 or more amino acids. Each heavy chain is comprised of a heavy chain variable region (VH) and a heavy chain constant region (CH). The heavy chain constant region is comprised of three domains, CH1, CH2 and CH3. Each light chain is comprised of a light chain variable region (VL) and a light chain constant region (CL). The light chain constant region is comprised of one domain, CL. The constant regions of the antibodies can mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (Clq) of the classical complement system.

[0096] The term "variable region" or "variable domain" as used herein refers to the domain of an antibody heavy or light chain that is involved in binding of the antigen-binding molecule to an antigen. The variable domains of the heavy chain and light chain (VH and VL, respectively) of a native antibody generally have similar structures, with each domain comprising four conserved framework regions (FRs) and three hypervariable regions (HVRs). A single VH or VL domain can be sufficient to confer antigen-binding specificity.

[0097] The term "variable" as used herein refers to certain segments of the variable domain that are universally different in sequence between antibodies. The V domain mediates antigen binding and defines the specificity of a particular antibody for its particular antigen. However, the variability is not evenly distributed throughout the variable domains. Instead, it is concentrated in three segments called hypervariable regions (HVRs) within the variable domains of both the light and heavy chains. The more highly conserved portions of variable domains are called the framework regions (FR). The variable domains of the heavy and light chains each comprise four FR regions, largely adopting a beta-sheet configuration, connected by three HVRs, which form loops connecting, and in some cases forming part of, the beta-sheet structure. The HVRs in each chain are held together in part by the FR regions of that chain, and, in part, by the HVRs of the other chain, to form the antibody's antigen binding site (see Kabat et al., Sequences of Immunological Interest, 5th Ed. National Institute of Health, Bethesda, MD (1991)). The constant domains are not involved directly in binding of an antibody to an antigen, but exhibit other effector functions, such as participation of the antibody in antibody dependent cellular cytotoxicity.

[0098] The term "hypervariable region" or "HVR", as used herein, refers to the regions of an antibody variable domain that are hypervariable in sequence and / or form structurally defined loops ("hypervariable loops") that are believed to be involved in binding the antigen. Generally, native four-chain antibodies comprise six HVRs; three in the VHand three in the VL. The term "complementarity determining region" or "CDR", as used herein, are the regions of most concentrated variation in amino acids in the variable region, with three CDRs in each of VHand VL(HCDR1-HCDR3 and LCDR1-LCDR3). The HVRs generally comprise amino acid residues from a "hypervariable loop" and / or from a "complementarity determining region" (CDR) that are hypervariable in sequence and / or involved in antigen recognition.

[0099] "Framework" or "FR" refers to the variable domain residues other than the hypervariable region (HVR) residues. The FR of a variable domain is generally composed of four FR domains: FR1, FR2, FR3, and FR4. Thus, the HVR and FR sequences generally appear in the following sequence in VHand (or VL): FR1-H1 (L1)-FR2-H2 (L2)-FR3-H3 (L3)-FR4.

[0100] The "class" of an antibody refers to the type of constant domain or constant region of the heavy chain possessed by an antibody. There are five classes of antibodies: IgA, IgD, IgE, IgG, and IgM and several of these can be further divided into subclasses (isotypes), e.g., IgGi, IgG2, IgG3, IgG4, IgAi and IgA2. The heavy chain constant domains that correspond to the different classes of immunoglobulins are called a, d, e, g, and m, respectively.

[0101] A "humanized antibody" comprises amino acid residues from non-human HVRs and amino acid residues from human FRs. In certain embodiments, a humanized antibody will comprise at least one, and typically two, variable domains in which all or substantially all HVRs (e.g., CDRs) correspond to those of a non-human antibody, and all or substantially all FRs correspond to those of a human antibody. A humanized antibody optionally can comprise at least a portion of an antibody constant region derived from a human antibody. An "antibody in a humanized form" of a non-human antibody, for example, refers to an antibody that has undergone humanization.

[0102] A "humanized antibody" has an amino acid sequence corresponding to that of an antibody produced by a human or a human cell or derived from an antibody of non-human origin that utilizes a human antibody repertoire or other human antibody-encoding sequences. This definition specifically excludes a humanized antibody comprising non-human antigen binding residues.

[0103] The present application also relates to amino acid sequence variants, which can be prepared by introducing appropriate modifications into the nucleotide sequence encoding the molecule, or by peptide synthesis. Such modifications include, for example, deletions from, insertions into, and / or substitutions of residues within the amino acid sequence of the antibody. Any combination of deletion, insertion, and substitution is made to arrive at the final construct, provided that the final construct possesses the desired characteristics, e.g., antigen binding activity. Sites of interest for substitutional

[0104] The term "polynucleotide" or "nucleic acid" or "nucleotide sequence" as used herein refers to an isolated nucleic acid molecule or construct, such as messenger RNA (mRNA), virus-derived RNA or plasmid DNA (pDNA). A polynucleotide can comprise conventional phosphodiester bonds or non-conventional bonds (e.g., amide, baseborate, etc.). The term "nucleic acid molecule" refers to any one or more nucleic acid segments, e.g., DNA or RNA fragments, present in a polynucleotide.

[0105] An "antibody fragment" or "antigen binding fragment" comprises a portion of an intact antibody. Examples of antibody fragments include but are not limited to Fab, Fab', F(ab')2, and Fv; diabodies, triabodies, tetrabodies, cross-Fab fragments; linear antibodies; single-chain antibody molecules (e.g., scFv); multispecific antibodies formed from antibody fragments and single domain antibodies (dAbs).

[0106] The term "antigen binding domain" or "antigen binding site" as used herein refers to the part of an antigen binding molecule that specifically binds to an antigenic determinant. More specifically, the term "antigen binding domain" refers to the part of an antibody that comprises the region that specifically binds to and is complementary to a part or the whole of an antigen. In the case of an antigen molecule that is large, the antigen binding molecule can bind only to a specific part of the antigen, which is referred to as an epitope. The antigen binding domain can be provided by, for example, one or more variable domains (also referred to as variable regions). Preferably, the antigen binding domain comprises an antibody light chain variable region (VL) and an antibody heavy chain variable region (VH). In one aspect, the antigen binding domain is capable of binding to its antigen and blocking or partially blocking the function of said antigen.

[0107] The term "antigenic determinant" as used herein is synonymous with "antigen" and "epitope" and refers to a site on a polypeptide macromolecule (e.g., a contiguous stretch of amino acids or a conformational configuration composed of different regions of non-contiguous amino acids) to which an antigen binding moiety binds, thereby forming an antigen binding moiety-antigen complex. Antigenic determinants can be found, for example, on the surface of tumor cells, microbe-infected cells, other diseased cells, immune cells, free agents in serum, and / or extracellular matrix (ECM). Unless otherwise specified, proteins used as antigens in the present application can be any native form of protein of any vertebrate origin, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The antigen can also be a human protein, or the antigen is a "full-length," unprocessed protein, as well as any form of the protein produced upon intracellular processing, or a naturally occurring variant of the protein, such as a splice variant or allelic variant.

[0108] The term "specifically binds to" as used herein refers to binding selectivity for an antigen and can be distinguished from non-specific or nonspecific binding. The term "affinity" or "binding affinity" as used herein refers to the strength of the noncovalent interaction between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Binding affinity can generally be expressed as a dissociation constant (Kd), which is the ratio of the dissociation rate constant to the association rate constant (K off and K on) can include different rate constants, as long as the ratio of the rate constants remains the same. The ability of an antigen binding molecule to bind to a particular antigen can be measured by enzyme linked immunosorbent assay (ELISA) or other techniques familiar to one skilled in the art, such as surface plasmon resonance (SPR) techniques, as well as traditional binding assays. In one embodiment, the extent of binding of an antigen binding molecule to an unrelated protein is less than about 10% of the binding of the antigen binding molecule to the antigen, as measured, for example, by SPR. In certain embodiments, the dissociation constant (Kd) of a molecule that binds to an antigen is <1 μM, <100 nM, <10 nM, <1 nM, <0.1 nM, <0.01 nM, or <0.001 nM (e.g., 10 -7 M or lower, e.g., 10 -7 M to 10 -13 M, e.g., 10 -9 M to 10 -13 M). The term "affinity" or "binding affinity" as used herein refers to the strength of the noncovalent interaction between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Binding affinity can be generally expressed as a dissociation constant (Kd), which is the ratio of the dissociation rate constant to the association rate constant, with smaller Kd values indicating smaller dissociation and representing stronger affinity between the antibody and antigen.

[0109] The term "isolated" nucleic acid molecule or polynucleotide as used herein refers to a nucleic acid molecule, DNA or RNA, that has been separated from a naturally occurring environment. A recombinant polynucleotide encoding a polypeptide comprised in a vector is also isolated in the present application. Other examples of isolated polynucleotides include a recombinant polynucleotide in a host cell, or a purified polynucleotide in solution. An isolated polynucleotide includes a polynucleotide molecule that is present in the cell in which it usually is contained, but the polynucleotide molecule is present extrachromosomally or at a chromosomal location that is different from its natural chromosomal location. Isolated RNA molecules include in vivo or in vitro RNA transcripts of the present application, in both positive and negative sense, and double-stranded forms. Isolated polynucleotides or nucleic acids of the present application further include synthetically produced such molecules. In addition, the polynucleotide or nucleic acid can be or can include regulatory elements, such as promoters, ribosome binding sites, or transcription terminators.

[0110] The term "vector" or "expression vector" as used herein refers to a DNA molecule that directs the expression of a particular gene operably linked thereto when introduced into a target cell. The vector includes vectors that are self-replicating nucleic acid structures as well as vectors that are incorporated into the genome of a host cell into which they have been introduced. The expression vectors of the present application comprise an expression cassette. The expression vector can undergo transcription of a large number of stable mRNAs. Once the expression vector is within the target cell, the ribonucleic acid molecule or protein encoded by the gene is produced by the cellular transcription and / or translation machinery. The term "expression cassette" as used herein refers to a recombinantly or synthetically generated polynucleotide having a series of nucleic acid elements that allow for the transcription of a particular nucleic acid in a target cell. The recombinant expression cassette portion of an expression vector typically includes, among other sequences, a nucleic acid sequence to be transcribed and a promoter.

[0111] The term "host cell" as used herein refers to a cell into which a foreign nucleic acid has been introduced, and also includes the progeny of such a cell. Host cells include "transformants" and "transformed cells," including primary transformed cells as well as progeny derived therefrom. The nucleic acid of the progeny can not be identical to that of the parent cell from which it was derived, but can contain mutations. Host cells are any type of cell that can be used to produce an antibody or antigen-binding fragment thereof of the present application. Host cells include cultured cells, e.g., cultured mammalian cells, such as CHO cells, HEK293 cells, BHK cells, NSO cells, SP2 / 0 cells, YO myeloma cells, P3X63 mouse myeloma cells, PER cells, PER.C6 cells, or hybridoma cells, yeast cells, insect cells, and plant cells, and also include cells contained within a transgenic animal, transgenic plant, or cultured plant or animal tissue.

[0112] The term "antibody drug conjugate" or "ADC" as used herein refers to a binding protein (such as an antibody or antigen-binding fragment thereof) chemically linked to one or more chemical drugs. In preferred embodiments, an ADC comprises a binding protein, a drug, and a linker linking the binding protein to the drug.

[0113] An "effective amount" of a drug refers to the amount necessary to bring about a desired physiological change in the cells or tissues receiving the administration. An "effective amount" includes an amount sufficient to ameliorate or prevent a symptom or condition of a medical disorder. An effective amount also means an amount sufficient to allow or facilitate diagnosis. An effective amount for a particular patient or veterinary subject can vary depending on factors such as the condition to be treated, the overall health status of the patient, the method route and dose of administration, and the severity of side effects. An effective amount can be the maximum dose or administration regimen that avoids significant side effects or toxic effects.

[0114] A "therapeutically effective amount" of a drug (e.g., a pharmaceutical composition) refers to the amount necessary to effect a desired therapeutic or prophylactic effect at a dose and interval and time of administration. For example, a therapeutically effective amount of a drug eliminates, reduces, delays, minimizes, or prevents adverse effects of a disease.

[0115] The term "individual" or "subject" refers to a mammal. Mammals include, but are not limited to, domesticated animals (e.g., cows, sheep, cats, dogs, and horses), primates (e.g., humans and non-human primates such as monkeys), rabbits, and rodents (e.g., mice and rats). Specifically, the individual or subject is a human.

[0116] The term "pharmaceutical composition" means a mixture of one or more antibodies of the disclosure or antibody or antigen-binding fragment thereof thereof with other chemical components such as physiologically / pharmaceutically-acceptable carriers or excipients. The purpose of a pharmaceutical composition is to facilitate administration of the active ingredient to a subject and to facilitate absorption of the active ingredient to thereby elicit the biological activity.

[0117] The term "pharmaceutically acceptable carrier" refers to a component of a pharmaceutical composition other than the active ingredient that is not toxic to a subject. Pharmaceutically acceptable excipients include, but are not limited to, buffers, stabilizers, and / or preservatives.

[0118] The term "treatment" refers to administration of an internal or external therapeutic agent, such as a composition comprising any of the antibodies of the disclosure or antibody or antigen-binding fragment thereof thereof or a nucleic acid molecule encoding the antibody or antibody or antigen-binding fragment thereof thereof, to a patient having one or more diseases or symptoms, which therapeutic agent has a therapeutic effect on these diseases or symptoms. Typically, the therapeutic agent is administered in an amount effective to alleviate one or more diseases or symptoms in a treated patient or population to induce regression of such symptoms or to inhibit development of such symptoms to any clinically measurable extent.

[0119] Embodiments of the present application will be described in detail below with reference to the examples.

[0120] The experimental methods used in the following examples are routine methods unless otherwise specified.

[0121] The reagents and / or kits used in the following examples are commercially available or can be synthesized by known methods.

[0122] Materials and Methods: 1. Cell Culture HEK293F cells (Thermo Fisher Scientific) were cultured in suspension in OPM-293 serum-free media (OPM Biosciences) in shake flasks at 37 °C, 8% CO2, 135 rpm. Cell density was reached at 4 × 10 6HEK293T cells (Thermo Fisher Scientific) were cultured adherently in Dulbecco's Modified Eagle Medium (DMEM) containing 10% fetal bovine serum (FBS) at 37 °C in a 5% CO2 incubator. After the cells reached approximately 80% confluence, they were digested with 0.25% trypsin, and after the cells were observed to detach, 4 mL of complete medium was immediately added to terminate the reaction. After the cells were mixed gently and centrifuged at 1,000 rpm for 5 minutes at room temperature, the supernatant was discarded, and the cells were resuspended in medium and subcultured at a ratio of 1:5.

[0123] 2. Expression and purification of rabies G protein Molecular construction: The G protein sequences (including signal peptides) of six rabies virus strains (P08667.1, CAI43218.1, ADM32132.1, ACR39382.1, ADJ29911.1, and ADD84785.1) were selected and artificially synthesized after codon optimization. A Twin-Strep-tag II and an 8-His-tag were added to the C-terminus of each sequence to facilitate purification and detection. The synthesized fragments were digested and ligated to the pCAGGS vector, and after sequencing verification, the vector was used for expression.

[0124] Cell transfection and protein expression: HEK 293F cells were counted and viability tested to ensure a cell viability of ≥95%. The cells were diluted to 1.7 - 2 × 10 6 Each of the two 10 mL OPM-293 culture media was added to a sterile 50 mL centrifuge tube, and 1.5 mg of PEI and 0.5 mg of the above plasmid DNA were added dropwise, and mixed gently. Then, the PEI-containing medium was added dropwise to the plasmid-containing solution, mixed well, and incubated at room temperature for 20 minutes to form a complex. The DNA / PEI complex was added dropwise to 500 mL of the diluted 293F cell suspension, mixed well, and incubated at 37 °C, 8% CO2, and 135 rpm. After 3 - 4 days of culture, the cells were collected for protein purification.

[0125] Protein purification: Transfected cultured 293F cells were collected by centrifugation at 4 °C, 3,800 rpm for 20 minutes. Cell pellets were resuspended in 20 mL lysis buffer (20 mM HEPES, 150 mM NaCl) and broken by repeated grinding. The resulting suspension was centrifuged at 4 °C, 36,000 rpm for 30-45 minutes, and the supernatant was discarded. The resulting pellet was resuspended in solubilization buffer (20 mM HEPES, 150 mM NaCl, 1% n-octyl-β-D-glucoside, containing protease inhibitors), and the suspension was thoroughly dispersed before being solubilized at 4 °C for 3 hours with magnetic stirring. The suspension was then centrifuged again at 4 °C, 36,000 rpm for 30-45 minutes, and the supernatant was collected for subsequent purification steps. The supernatant was thoroughly incubated with a Strep-Tactin affinity chromatography column, and the non-specifically bound impurities were washed away with protein buffer (20 mM HEPES, 150 mM NaCl, 1% n-octyl-β-D-glucoside). Finally, the target protein was eluted using protein elution buffer (50 mM Biotin, 20 mM HEPES, 150 mM NaCl, 1% n-octyl-β-D-glucoside), and the protein was identified by SDS-PAGE and peptide mapping. The target membrane protein was obtained and used for downstream experiments.

[0126] Example 1 Obtaining of antibodies 1. Isolation of human peripheral blood mononuclear cells (PBMCs) , PBMC isolation Vaccinees with different immune backgrounds (all volunteers had no chronic disease basis, and their ages ranged from 23 to 41 years old, and they were inoculated with PV / PM / CTN rabies vaccine according to a 4-needle immunization program or a 5-needle immunization program) were collected in an anticoagulant blood bag, stored and transported at 4 °C. 15 mL Ficoll-Paque PLUS density gradient medium (cytiva) was added to a lymphocyte separation tube (Sigma-Aldrich), and centrifuged at 400 g for 5 minutes at room temperature. Then, about 25 mL of anticoagulant whole blood was slowly added, and centrifuged at 900 g for 15 minutes at room temperature, with the lifting speed of the centrifuge being the lowest. After centrifugation, a flat-bar pasting pipette was gently inserted into the mononuclear cell layer, and the layer was carefully transferred to a 50 mL centrifuge tube by rotating along the tube wall. D-Hanks buffer (Solarbio) without calcium and magnesium was added to a total volume of 45 mL, and centrifuged at 400 g for 10 minutes to remove residual plasma. If there was red blood cell contamination, ACK red blood cell lysis buffer (150 mM NH4Cl, 10 mM KHCO3, 0.1 mM CaCl2) was added to a final concentration of 0.8%, and the mixture was incubated at room temperature for 5 minutes. The mixture was then centrifuged at 400 g for 10 minutes to remove the red blood cells. The supernatant was collected and centrifuged at 400 g for 10 minutes to remove the residual plasma. The resulting pellet was resuspended in D-Hanks buffer (Solarbio) to a final volume of 1 mL, and the concentration of PBMCs was determined by hemocytometer counting. 10 H16 N2O8) and resuspended to 45 mL with D-Hanks. The cells were centrifuged for 10 minutes. The final resuspension was done with D-Hanks, and the aggregated cells were removed by filtration. The cells were counted and stored at 4°C. The cells were resuspended in freezing medium (fetal bovine serum and DMSO in a 9:1 ratio) and aliquoted into labeled cryotubes, which were placed in a programmed cooling freezing box at -80°C overnight and then transferred to liquid nitrogen for long-term storage.

[0127] 2. Single specific memory B cells sorting for RABV G antigen binding To obtain human memory B cells specific for rabies virus G protein (RABV-G), PBMCs were sorted by multicolor flow cytometry. PBMCs were incubated with the following fluorescently labeled antibodies: PE / Cyanine7-labeled anti-human CD19 antibody (BioLegend) to identify B lymphocytes, PE-labeled anti-human IgG antibody and PerCP / Cyanine5.5-labeled human IgM / D antibody to screen for IgG-positive, IgM / D-negative B cells, and APC-labeled anti-human CD27 antibody to identify memory B cells. Rabies virus G protein was labeled with Alexa Fluor 488 (BioLegend) to identify antigen-specific B cells. After staining, cell sorting was performed using a BD FACSAria III flow cytometer (BD Biosciences). The final obtained RABV G protein-binding single CD19 + IgM / IgD - IgG + CD27 + Memory B cells were sorted into 96-well PCR plates for subsequent single-cell RT-PCR amplification of antibody variable region sequences.

[0128] 3. Antibody gene cloning and transient expression The sorted single memory B cells were subjected to single-cell level antibody gene amplification. First-strand cDNA synthesis was performed, followed by amplification of full-length cDNA and variable region fragments of IgG heavy and light chains, respectively. Based on this, IgG heavy and light chain expression frames (transcriptional-active DNA, TAD) containing CMV promoters and polyA tails were constructed by fragment splicing reactions, respectively. The obtained IgG heavy and light chain-containing TADs were co-transfected into HEK293T cells. After 72 hours of transfection, the culture supernatant was collected, which was the antibody expression product, and was used for subsequent binding ability and neutralization function detection.

[0129] Example 2 Primary screening of positive candidate antibodies The cell supernatant (antibody expression product) obtained in Example 1 was used for functional primary screening in pseudovirus neutralization experiment. The neutralization experiment used the rabies virus VSV pseudovirus system to evaluate the neutralization ability of the antibody to representative rabies virus strains.

[0130] 1. Preparation of rabies virus pseudovirus The rabies virus G protein sequence (ADM32132.1, ACR39382.1, ADJ29911.1, ADD84785.1, CAI43218.1, P08667.1) was synthesized after codon optimization and constructed in pcDNA3.1 plasmid, and the plasmid was amplified. The plasmid was transfected into HEK293T with a confluence of 70-80%. After 24 hours, the TCID 50 was 7x10 4 The vesicular stomatitis virus pseudovirus (G ΔG luciferase VSV pseudovirus) was diluted with 2% FBS DMEM and added to the cells, and incubated in a 37°C, 5% CO2 incubator for 3-5 hours. After washing with PBS solution, the cell culture fluid was replaced with new 2% FBS DMEM medium, and incubated in a 37°C, 5% CO2 incubator for 24 hours. The culture supernatant was centrifuged at 1,000 rpm and 5 minutes at room temperature, and the supernatant was aliquoted and stored in a -80°C refrigerator.

[0131] 2. Antibody neutralization experiment based on rabies virus pseudovirus The antibody to be tested (the supernatant of the transient expression cells obtained in Example 1) was initially diluted 30 times with DMEM, and then serially diluted at a ratio of 5 times, with a total of 8 dilution gradients. The dilution operation was carried out in a 96-well plate, and cell controls (100 μL DMEM) and virus controls (50 μL pseudovirus and 50 μL DMEM) were set up. Each group of dilutions and diluted pseudovirus were incubated at 37°C, 5% CO2 for 1 hour. Then, the HEK293T cells in good growth condition were digested and adjusted to a concentration of 4x10 5 cells / mL, and 100 μL of cell suspension was added to each well. Incubation was continued in a 37°C, 5% CO2 incubator for 24 hours. After 24 hours, the supernatant was discarded, and 100 μL of ONE-Glo™ EX Reagent (Promega) was added, and the reaction was carried out in the dark for 2 minutes. The fluorescence signal (RLU) was read using a PerkinElmer EnSight multifunctional imaging enzyme labeler to evaluate the neutralization effect. The neutralization curve was plotted according to the RLU change, and the IC 50 value was calculated.

[0132] Figure 1The results showed that part of the expression products showed good neutralization activity, thereby preliminarily screening a batch of positive candidate antibodies.

[0133] Example 3 Construction of antibody expression vector According to the supernatant screened in Example 2 and the remarkable effect, the heavy chain and light chain variable region sequencing (antibody sequence is shown in Tables 1-6) was performed, and the naturally paired variable region sequences were synthesized. The heavy chain and light chain were respectively cloned into the expression vector pcDNA3.1 containing human IgG1 constant region, and the plasmid was amplified. The HEK293F cells were used for transient transfection expression, and the cells were collected after 5 days of transfection at 4 ℃, 3,800 rpm for 40 minutes. The cell precipitate was discarded, the supernatant was collected and filtered (0.45 μm filter). The filtered supernatant was incubated with a protein A affinity chromatography column at 4 ℃ for 1 hour, and after washing away the non-specifically bound impurities with PBS solution, the target antibody was eluted with 0.1 M glycine solution at pH = 2.5, and immediately neutralized with 1 M Tris solution at pH = 8.0. The purified antibody was verified by SDS-PAGE and used for subsequent functional verification.

[0134] Example 4 Enzyme-linked immunosorbent assay (ELISA) evaluation The purified rabies virus G protein P08667.1, CAI43218.1, ACR39382.1 was diluted to 2 μg / mL with PBS buffer, coated on a 96-well ELISA plate, and incubated at 4 ℃ overnight. 3 times of PBST (PBS containing 0.05% Tween-20) washing, 5% skim milk powder blocking solution was added to each well, and the room temperature was blocked for 2 hours. Then the antibody to be tested (positive candidate antibody obtained in Example 2) was added, diluted and incubated at room temperature for 1 hour. After washing the plate, 5000-fold diluted HRP-labeled anti-human IgG antibody (Sigma-Aldrich) was added, and incubated at room temperature for 1 hour. After washing the plate again, TMB color developing liquid (Bi Yun Tian) was added and reacted for about 10 minutes. After stopping the reaction with a stopping solution, the absorbance value was read at 450 nm wavelength.

[0135] Figure 2 The ELISA results showed that most of the antibodies (such as #230, #272, #249, #296, #213 and #253) had good binding ability to the G protein of the classical vaccine strain (P08667.1, CAI43218.1, ACR39382.1), and some antibodies showed broad-spectrum binding ability among different strains.

[0136] Example 5 Rabies virus pseudovirus-based antibody neutralization experiment 1. Preparation of rabies virus pseudovirus Rabies virus G protein sequence (GenBank: ADM32132.1, ACR39382.1, ADJ29911.1, ADD84785.1, CAI43218.1, P08667.1) was synthesized after codon optimization and constructed in pcDNA3.1 plasmid, and the plasmid was amplified. The plasmid was transfected into HEK293T at 70-80% confluence. After 24 hours, the TCID 50 was 7x10 4 Vesicular stomatitis virus pseudovirus (G ΔG luciferase VSV pseudovirus) was diluted with 2% FBS DMEM and added to the cells, and incubated at 37°C, 5% CO2 incubator for 3-5 hours. After washing with PBS solution, the cell culture fluid was replaced with new 2% FBS DMEM medium, and continued to be placed in a 37°C, 5% CO2 incubator for 24 hours. The culture supernatant was centrifuged at 1,000 rpm for 5 minutes at room temperature, and the supernatant was aliquoted and stored in a -80°C refrigerator.

[0137] 2. Antibody neutralization experiment based on rabies virus pseudovirus The antibody to be tested (positive candidate antibody obtained in Example 2) was initially diluted 30 times with DMEM, and then serially diluted at a ratio of 3 or 5 times, a total of 8 dilution gradients were set. The dilution operation was carried out in a 96-well plate, and cell controls (100 μL DMEM) and virus controls (50 μL pseudovirus and 50 μL DMEM) were set up. Each group of dilutions and diluted pseudovirus were incubated at 37°C, 5% CO2 for 1 hour. Then, the HEK293T cells in good growth condition were digested and adjusted to a concentration of 4x10 5 cells / mL, 100 μL of cell suspension was added to each well. Continue to incubate at 37°C, 5% CO2 incubator for 24 hours. After 24 hours, the supernatant was discarded, and 100 μL of ONE-Glo™ EX Reagent (Promega) was added, and the reaction was carried out in the dark for 2 minutes. The fluorescence signal (RLU) was read using PerkinElmer EnSight multifunctional imaging enzyme labeler to evaluate the neutralization effect. According to the change of RLU, the neutralization curve was drawn, and the IC 50 value was calculated.

[0138] Figure 3 The results show that some antibodies (such as #230, #272, #249, #296, #213 and #253) have strong neutralization ability to multiple strains, suggesting their potential as broad-spectrum neutralizing antibodies. The heavy chain variable region, light chain variable region and CDR sequences of antibodies #230, #272, #249, #296, #213 and #253 are shown in Tables 1-6.

[0139] Table 1

[0140] Table 2

[0141] Table 3

[0142] Table 4

[0143] Table 5

[0144] Table 6

[0145] Example 6 Determination of antigen-antibody affinity dissociation constant The affinity between purified antibodies and rabies virus G protein was assessed using biolayer interferometry (BLI).

[0146] The purified G protein (P08667.1) was diluted to 20 μg / mL, and the initial concentration of the candidate antibody was 100 nM, followed by 2-fold serial dilutions. Experiments were performed using an Octet Red96 sensor. The G protein was immobilized using a Ni-NTA sensor, and a program was set to sequentially detect affinity dissociation of each candidate antibody screened in Example 5. Between each detection round, the sensor was regenerated using a glycine solution at pH 1.5, followed by neutralization with 10 mM NiSO4 solution. The affinity dissociation curves were fitted using the Octet system's Data Analysis software, and the affinity dissociation constant was calculated to evaluate the antibody binding activity.

[0147] The results are as follows Figure 4 As shown, antibodies #230, #272, #249, #296, #213, and #253 exhibit good affinity and binding activity to the rabies virus G protein.

[0148] Example 7: Neutralization Experiment of Pseudovirus Antibody Based on Street Rabies Virus To verify the broad-spectrum neutralizing ability of candidate antibodies in real infection scenarios, this embodiment constructed a pseudovirus neutralization screening system containing 46 RABV street viruses.

[0149] The antibody to be tested (monoclonal antibody determined in Example 5) and antibody combinations (antibody mass concentration 1:1 combination) were diluted 30 times initially with DMEM, then serially diluted at a ratio of 3 or 5 times, a total of 8 dilution gradients were set. The dilution operation was carried out in a 96-well plate, and cell controls (100 μL DMEM) and virus controls (50 μL pseudovirus and 50 μL DMEM) were set up. Each group of dilutions and diluted pseudovirus were incubated at 37 °C, 5% CO2 for 1 hour. Then, the well-grown HEK293T cells were digested and adjusted to a concentration of 4 × 10 5 cells / mL, 100 μL of cell suspension was added to each well. Continue to incubate at 37 °C, 5% CO2 incubator for 24 hours. After 24 hours, discard the supernatant, add 100 μL ONE-Glo™ EX Reagent (Promega), avoid light reaction for 2 minutes. Use PerkinElmer EnSight multifunctional imaging enzyme labeler to read the fluorescence signal (RLU) to evaluate the neutralization effect. According to the change of RLU, draw the neutralization curve, calculate the IC 50 value.

[0150] Figure 5 The experimental results show that most of the candidate antibodies (such as #230, #272, #249, #296, #213 and #253) retain the neutralization ability in the street virus background, and some antibody combinations (such as #230 / #272, #249 / #213, #296 / #213, #249 / #253, #296 / #253, #249 / #272) exhibit synergistic or complementary neutralization effect among strains.

[0151] Example 8 Antibody neutralization experiment based on lyssavirus pseudovirus To determine whether the candidate antibody combination has neutralization effect on some viruses of the lyssavirus genus, the neutralization activity of the antibodies on various lyssavirus strains (ABLV: KU739052.1, ARAV: EF614259.1, BBLV: KU761304.1, EBLV1: EU352768.1, EBLV2: AAX62813.1, IRKV: EF614260.1, KHUV: EF614261.1) was further tested according to the method of Example 7.

[0152] Figure 6 The results show that some antibody combinations (such as #230 / #272, #249 / #213, #296 / #213, #249 / #253, #296 / #253, #249 / #272) maintain high neutralization potency in various lyssavirus strains, suggesting that some combinations may exhibit potential broad-spectrum neutralization advantage.

[0153] The technical solution of the present application is not limited to the above specific embodiments, and any technical variation made according to the technical solution of the present application falls within the protection scope of the present application.

Claims

1. An antibody or antigen-binding fragment thereof that specifically binds to rabies virus, said antibody or antigen-binding fragment comprising: (1) The following three heavy chain variable region complementarity-determining regions (HCDRs): HCDR1, having the amino acid sequence of HCDR1 contained in the heavy chain variable region as shown in any one of SEQ ID NO: 1, 8, 15, 22, 29 and 36, or having one or more amino acid substitutions, deletions or additions compared to the amino acid sequence of HCDR1 contained in the heavy chain variable region. HCDR2, having the amino acid sequence of HCDR2 contained in the heavy chain variable region as shown in any one of SEQ ID NO: 1, 8, 15, 22, 29 and 36, or having one or more amino acid substitutions, deletions or additions compared to the amino acid sequence of HCDR2 contained in the heavy chain variable region. HCDR3, having the amino acid sequence of HCDR3 contained in the heavy chain variable region as shown in any one of SEQ ID NO: 1, 8, 15, 22, 29 and 36, or having one or more amino acid substitutions, deletions or additions compared to the amino acid sequence of HCDR3 contained in the heavy chain variable region; and (2) The following three light chain variable region complementarity-determining regions (LCDRs): LCDR1 having the amino acid sequence of LCDR1 contained in the light chain variable region as shown in any one of SEQ ID NO: 2, 9, 16, 23, 30 and 37, or having an amino acid sequence with one or more amino acid substitutions, deletions or additions compared to the amino acid sequence of LCDR1 contained in the light chain variable region. LCDR2 having the amino acid sequence of LCDR2 contained in the light chain variable region as shown in any one of SEQ ID NO: 2, 9, 16, 23, 30 and 37, or having an amino acid sequence with one or more amino acid substitutions, deletions or additions compared to the amino acid sequence of LCDR2 contained in the light chain variable region. LCDR3 having the amino acid sequence of LCDR3 contained in the light chain variable region as shown in any one of SEQ ID NO: 2, 9, 16, 23, 30 and 37, or having an amino acid sequence with one or more amino acid substitutions, deletions or additions compared to the amino acid sequence of LCDR3 contained in the light chain variable region. Preferably, the HCDR1-3 and / or the LCDR1-3 are defined by rules of Kabat, AbM, Chothia, Contact, IMGT or combinations thereof.

2. The antibody or its antigen-binding fragment according to claim 1, wherein, The antibody or its antigen-binding fragment comprises: As shown in SEQ ID NO: 1, the three HCDRs contained in the heavy chain variable region, or amino acid sequences having one or more amino acid substitutions, deletions, or additions compared to the amino acid sequences of the three HCDRs contained in the heavy chain variable region, or amino acid sequences having at least 80% sequence identity with them; and as shown in SEQ ID NO: 2, the three LCDRs contained in the light chain variable region, or amino acid sequences having one or more amino acid substitutions, deletions, or additions compared to the amino acid sequences of the three LCDRs contained in the light chain variable region, or amino acid sequences having at least 80% sequence identity with them; or As shown in SEQ ID NO: 8, the three HCDRs contained in the heavy chain variable region, or amino acid sequences having one or more amino acid substitutions, deletions, or additions compared to the amino acid sequences of the three HCDRs contained in the heavy chain variable region, or amino acid sequences having at least 80% sequence identity with them; and as shown in SEQ ID NO: 9, the three LCDRs contained in the light chain variable region, or amino acid sequences having one or more amino acid substitutions, deletions, or additions compared to the amino acid sequences of the three LCDRs contained in the light chain variable region, or amino acid sequences having at least 80% sequence identity with them; or As shown in SEQ ID NO: 15, the three HCDRs contained in the heavy chain variable region, or amino acid sequences having one or more amino acid substitutions, deletions, or additions compared to the amino acid sequences of the three HCDRs contained in the heavy chain variable region, or amino acid sequences having at least 80% sequence identity with them; and as shown in SEQ ID NO: 16, the three LCDRs contained in the light chain variable region, or amino acid sequences having one or more amino acid substitutions, deletions, or additions compared to the amino acid sequences of the three LCDRs contained in the light chain variable region, or amino acid sequences having at least 80% sequence identity with them; or As shown in SEQ ID NO: 22, the three HCDRs contained in the heavy chain variable region, or amino acid sequences having one or more amino acid substitutions, deletions, or additions compared to the amino acid sequences of the three HCDRs contained in the heavy chain variable region, or amino acid sequences having at least 80% sequence identity with them; and as shown in SEQ ID NO: 23, the three LCDRs contained in the light chain variable region, or amino acid sequences having one or more amino acid substitutions, deletions, or additions compared to the amino acid sequences of the three LCDRs contained in the light chain variable region, or amino acid sequences having at least 80% sequence identity with them; or As shown in SEQ ID NO: 29, the three HCDRs contained in the heavy chain variable region, or amino acid sequences having one or more amino acid substitutions, deletions, or additions compared to the amino acid sequences of the three HCDRs contained in the heavy chain variable region, or amino acid sequences having at least 80% sequence identity with them; and as shown in SEQ ID NO: 30, the three LCDRs contained in the light chain variable region, or amino acid sequences having one or more amino acid substitutions, deletions, or additions compared to the amino acid sequences of the three LCDRs contained in the light chain variable region, or amino acid sequences having at least 80% sequence identity with them; or The three HCDRs contained in the heavy chain variable region as shown in SEQ ID NO: 36, or amino acid sequences having one or more amino acid substitutions, deletions, or additions compared to the amino acid sequences of the three HCDRs contained in the heavy chain variable region, or amino acid sequences having at least 80% sequence identity with them; and the three LCDRs contained in the light chain variable region as shown in SEQ ID NO: 37, or amino acid sequences having one or more amino acid substitutions, deletions, or additions compared to the amino acid sequences of the three LCDRs contained in the light chain variable region, or amino acid sequences having at least 80% sequence identity with them. Preferably, the antibody or its antigen-binding fragment comprises: HCDR1, having an amino acid sequence as shown in any one of SEQ ID NO: 3, 10, 17, 24, 31 or 38, an amino acid sequence having one or more amino acid substitutions, deletions or additions compared to the HCDR1, or an amino acid sequence having at least 80% sequence identity with the HCDR1; and / or HCDR2, having an amino acid sequence as shown in any one of SEQ ID NO: 4, 11, 18, 25, 32 or 39, an amino acid sequence having one or more amino acid substitutions, deletions or additions compared to the HCDR2, or an amino acid sequence having at least 80% sequence identity with the HCDR2; and / or HCDR3, having an amino acid sequence as shown in any one of SEQ ID NO: 5, 12, 19, 26, 33 or 40, having an amino acid sequence with one or more amino acid substitutions, deletions or additions compared to the HCDR3, or having an amino acid sequence with at least 80% sequence identity to the HCDR3; and / or LCDR1, having an amino acid sequence as shown in any one of SEQ ID NO: 6, 13, 20, 27, 34 or 41, an amino acid sequence having one or more amino acid substitutions, deletions or additions compared to the LCDR1, or an amino acid sequence having at least 80% sequence identity with the LCDR1; and / or LCDR2, having an amino acid sequence as shown in WAS, ASS, GAS, EVT, EDN, or AAS, or having an amino acid sequence with a substitution, deletion, or addition of one amino acid compared to it; and / or LCDR3, having an amino acid sequence as shown in any one of SEQ ID NO: 7, 14, 21, 28, 35 or 42, an amino acid sequence having one or more amino acid substitutions, deletions or additions compared to the LCDR3, or an amino acid sequence having at least 80% sequence identity with the LCDR3. More preferably, the antibody or its antigen-binding fragment comprises: HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, or amino acid sequences having one or more amino acid substitutions, deletions or additions compared to the amino acid sequences of HCDR1, HCDR2 and HCDR3, or amino acid sequences having at least 80% sequence identity with them, and / or LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 6, with amino acid sequences of WAS and SEQ ID NO: 7, or amino acid sequences having one or more amino acid substitutions, deletions or additions compared to the amino acid sequences of LCDR1, LCDR2 and LCDR3, or amino acid sequences having at least 80% sequence identity with them; More preferably, the antibody or its antigen-binding fragment comprises: HCDR1, HCDR2, and HCDR3 as shown in SEQ ID NO: 10, SEQ ID NO: 11, and SEQ ID NO: 12, or amino acid sequences having one or more amino acid substitutions, deletions, or additions compared to the amino acid sequences of HCDR1, HCDR2, and HCDR3, or amino acid sequences having at least 80% sequence identity with them, and / or LCDR1, LCDR2, and LCDR3 as shown in SEQ ID NO: 13, with amino acid sequences of ASS and SEQ ID NO: 14, or amino acid sequences having one or more amino acid substitutions, deletions, or additions compared to the amino acid sequences of LCDR1, LCDR2, and LCDR3, or amino acid sequences having at least 80% sequence identity with them; More preferably, the antibody or its antigen-binding fragment comprises: HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 17, SEQ ID NO: 18 and SEQ ID NO: 19, or amino acid sequences having one or more amino acid substitutions, deletions or additions compared to the amino acid sequences of HCDR1, HCDR2 and HCDR3, or amino acid sequences having at least 80% sequence identity with them, and / or LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 20, with amino acid sequences of GAS and SEQ ID NO: 21, or amino acid sequences having one or more amino acid substitutions, deletions or additions compared to the amino acid sequences of LCDR1, LCDR2 and LCDR3, or amino acid sequences having at least 80% sequence identity with them; More preferably, the antibody or its antigen-binding fragment comprises: HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 24, SEQ ID NO: 25 and SEQ ID NO: 26, or amino acid sequences having one or more amino acid substitutions, deletions or additions compared to the amino acid sequences of HCDR1, HCDR2 and HCDR3, or amino acid sequences having at least 80% sequence identity with them, and / or LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 27, amino acid sequences EVT and SEQ ID NO: 28, or amino acid sequences having one or more amino acid substitutions, deletions or additions compared to the amino acid sequences of LCDR1, LCDR2 and LCDR3, or amino acid sequences having at least 80% sequence identity with them; More preferably, the antibody or its antigen-binding fragment comprises: HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 31, SEQ ID NO: 32 and SEQ ID NO: 33, or amino acid sequences having one or more amino acid substitutions, deletions or additions compared to the amino acid sequences of HCDR1, HCDR2 and HCDR3, or amino acid sequences having at least 80% sequence identity with them, and / or LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 34, amino acid sequences EDN and SEQ ID NO: 35, or amino acid sequences having one or more amino acid substitutions, deletions or additions compared to the amino acid sequences of LCDR1, LCDR2 and LCDR3, or amino acid sequences having at least 80% sequence identity with them; More preferably, the antibody or its antigen-binding fragment comprises: HCDR1, HCDR2, and HCDR3 as shown in SEQ ID NO: 38, SEQ ID NO: 39, and SEQ ID NO: 40, or amino acid sequences having one or more amino acid substitutions, deletions, or additions compared to the amino acid sequences of HCDR1, HCDR2, and HCDR3, or amino acid sequences having at least 80% sequence identity with them, and / or LCDR1, LCDR2, and LCDR3 as shown in SEQ ID NO: 41, with amino acid sequences AAS, and SEQ ID NO: 42, or amino acid sequences having one or more amino acid substitutions, deletions, or additions compared to the amino acid sequences of LCDR1, LCDR2, and LCDR3, or amino acid sequences having at least 80% sequence identity with them.

3. The antibody or its antigen-binding fragment according to claim 1, wherein, The antibody or its antigen-binding fragment comprises: The heavy chain variable region comprises an amino acid sequence as shown in SEQ ID NO: 1, 8, 15, 22, 29 or 36, an amino acid sequence having one or more amino acid substitutions, deletions or additions compared to it, or an amino acid sequence having at least 80% sequence identity with it; and / or The light chain variable region contains an amino acid sequence as shown in SEQ ID NO: 2, 9, 16, 23, 30 or 37, an amino acid sequence having one or more amino acid substitutions, deletions or additions compared to it, or an amino acid sequence having at least 80% sequence identity with it. Preferably, the antibody or its antigen-binding fragment comprises: a heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 1, an amino acid sequence having one or more amino acid substitutions, deletions or additions compared to the heavy chain, or an amino acid sequence having at least 80% sequence identity with the heavy chain variable region; and a light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 2, an amino acid sequence having one or more amino acid substitutions, deletions or additions compared to the light chain, or an amino acid sequence having at least 80% sequence identity with the light chain variable region. Preferably, the antibody or its antigen-binding fragment comprises: a heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 8, an amino acid sequence having one or more amino acid substitutions, deletions or additions compared to the heavy chain, or an amino acid sequence having at least 80% sequence identity with the heavy chain variable region; and a light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 9, an amino acid sequence having one or more amino acid substitutions, deletions or additions compared to the light chain, or an amino acid sequence having at least 80% sequence identity with the light chain variable region. Preferably, the antibody or its antigen-binding fragment comprises: a heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 15, an amino acid sequence having one or more amino acid substitutions, deletions or additions compared to the heavy chain, or an amino acid sequence having at least 80% sequence identity with the heavy chain variable region; and a light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 16, an amino acid sequence having one or more amino acid substitutions, deletions or additions compared to the light chain, or an amino acid sequence having at least 80% sequence identity with the light chain variable region. Preferably, the antibody or its antigen-binding fragment comprises: a heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 22, an amino acid sequence having one or more amino acid substitutions, deletions or additions compared to the heavy chain, or an amino acid sequence having at least 80% sequence identity with the heavy chain variable region; and a light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 23, an amino acid sequence having one or more amino acid substitutions, deletions or additions compared to the light chain, or an amino acid sequence having at least 80% sequence identity with the light chain variable region. Preferably, the antibody or its antigen-binding fragment comprises: a heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 29, an amino acid sequence having one or more amino acid substitutions, deletions or additions compared to the heavy chain, or an amino acid sequence having at least 80% sequence identity with the heavy chain variable region; and a light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 30, an amino acid sequence having one or more amino acid substitutions, deletions or additions compared to the light chain, or an amino acid sequence having at least 80% sequence identity with the light chain variable region. Preferably, the antibody or its antigen-binding fragment comprises: a heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 36, an amino acid sequence having one or more amino acid substitutions, deletions or additions compared to the heavy chain, or an amino acid sequence having at least 80% sequence identity with the heavy chain variable region; and a light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 37, an amino acid sequence having one or more amino acid substitutions, deletions or additions compared to the light chain, or an amino acid sequence having at least 80% sequence identity with the light chain variable region. Preferably, the antibody is of type IgA, IgD, IgE, IgG, or IgM; Preferably, the antibody or its antigen-binding fragment is scFv, Fab, Fab', (Fab')2, Fv fragment, dsFv, biantibody, bispecific antibody, or multispecific antibody; Preferably, the antibody or its antigen-binding fragment is a chimeric antibody, a human antibody, or a humanized antibody. Preferably, the antibody or its antigen-binding fragment further includes a heavy chain constant region and / or a light chain constant region; More preferably, the heavy chain constant region is selected from the heavy chain constant regions of IgG1, IgG2, IgG3 or IgG4.

4. An antibody composition comprising a first antibody and a second antibody, wherein the first antibody and / or the second antibody are selected from the antibodies or antigen-binding fragments thereof according to any one of claims 1-3; Preferably, the first antibody comprises: HCDR1, HCDR2, and HCDR3 as shown in SEQ ID NO: 10, SEQ ID NO: 11, and SEQ ID NO: 12, and LCDR1, LCDR2, and LCDR3 as shown in SEQ ID NO: 13 with the amino acid sequence ASS and SEQ ID NO: 14; and the second antibody comprises: HCDR1, HCDR2, and HCDR3 as shown in SEQ ID NO: 31, SEQ ID NO: 32, and SEQ ID NO: 33, and LCDR1, LCDR2, and LCDR3 as shown in SEQ ID NO: 34 with the amino acid sequence EDN and SEQ ID NO: 35; Preferably, the first antibody comprises: HCDR1, HCDR2, and HCDR3 as shown in SEQ ID NO: 17, SEQ ID NO: 18, and SEQ ID NO: 19, and LCDR1, LCDR2, and LCDR3 as shown in SEQ ID NO: 20 with the amino acid sequence GAS and SEQ ID NO: 21; and the second antibody comprises: HCDR1, HCDR2, and HCDR3 as shown in SEQ ID NO: 3, SEQ ID NO: 4, and SEQ ID NO: 5, and LCDR1, LCDR2, and LCDR3 as shown in SEQ ID NO: 6 with the amino acid sequence WAS and SEQ ID NO: 7; Preferably, the first antibody comprises: HCDR1, HCDR2, and HCDR3 as shown in SEQ ID NO: 38, SEQ ID NO: 39, and SEQ ID NO: 40, and LCDR1, LCDR2, and LCDR3 as shown in SEQ ID NO: 41 with amino acid sequences of AAS and SEQ ID NO: 42; and the second antibody comprises: HCDR1, HCDR2, and HCDR3 as shown in SEQ ID NO: 3, SEQ ID NO: 4, and SEQ ID NO: 5, and LCDR1, LCDR2, and LCDR3 as shown in SEQ ID NO: 6 with amino acid sequences of WAS and SEQ ID NO: 7; Preferably, the first antibody comprises: HCDR1, HCDR2, and HCDR3 as shown in SEQ ID NO: 17, SEQ ID NO: 18, and SEQ ID NO: 19, and LCDR1, LCDR2, and LCDR3 as shown in SEQ ID NO: 20 with the amino acid sequence GAS and SEQ ID NO: 21; and the second antibody comprises: HCDR1, HCDR2, and HCDR3 as shown in SEQ ID NO: 24, SEQ ID NO: 25, and SEQ ID NO: 26, and LCDR1, LCDR2, and LCDR3 as shown in SEQ ID NO: 27 with the amino acid sequence EVT and SEQ ID NO: 28; Preferably, the first antibody comprises: HCDR1, HCDR2, and HCDR3 as shown in SEQ ID NO: 38, SEQ ID NO: 39, and SEQ ID NO: 40, and LCDR1, LCDR2, and LCDR3 as shown in SEQ ID NO: 41 with amino acid sequences of AAS and SEQ ID NO: 42; and the second antibody comprises: HCDR1, HCDR2, and HCDR3 as shown in SEQ ID NO: 24, SEQ ID NO: 25, and SEQ ID NO: 26, and LCDR1, LCDR2, and LCDR3 as shown in SEQ ID NO: 27 with amino acid sequences of EVT and SEQ ID NO: 28; Preferably, the first antibody comprises: HCDR1, HCDR2, and HCDR3 as shown in SEQ ID NO: 17, SEQ ID NO: 18, and SEQ ID NO: 19, and LCDR1, LCDR2, and LCDR3 as shown in SEQ ID NO: 20 with the amino acid sequence GAS and SEQ ID NO: 21; and the second antibody comprises: HCDR1, HCDR2, and HCDR3 as shown in SEQ ID NO: 31, SEQ ID NO: 32, and SEQ ID NO: 33, and LCDR1, LCDR2, and LCDR3 as shown in SEQ ID NO: 34 with the amino acid sequence EDN and SEQ ID NO: 35; More preferably, the first antibody comprises a heavy chain variable region as shown in SEQ ID NO: 8 and a light chain variable region as shown in SEQ ID NO: 9, and the second antibody comprises a heavy chain variable region as shown in SEQ ID NO: 29 and a light chain variable region as shown in SEQ ID NO: 30; More preferably, the first antibody comprises a heavy chain variable region as shown in SEQ ID NO: 15 and a light chain variable region as shown in SEQ ID NO: 16, and the second antibody comprises a heavy chain variable region as shown in SEQ ID NO: 1 and a light chain variable region as shown in SEQ ID NO: 2; More preferably, the first antibody comprises a heavy chain variable region as shown in SEQ ID NO: 36 and a light chain variable region as shown in SEQ ID NO: 37, and the second antibody comprises a heavy chain variable region as shown in SEQ ID NO: 1 and a light chain variable region as shown in SEQ ID NO: 2; More preferably, the first antibody comprises a heavy chain variable region as shown in SEQ ID NO: 15 and a light chain variable region as shown in SEQ ID NO: 16, and the second antibody comprises a heavy chain variable region as shown in SEQ ID NO: 22 and a light chain variable region as shown in SEQ ID NO: 23; More preferably, the first antibody comprises a heavy chain variable region as shown in SEQ ID NO: 36 and a light chain variable region as shown in SEQ ID NO: 37, and the second antibody comprises a heavy chain variable region as shown in SEQ ID NO: 22 and a light chain variable region as shown in SEQ ID NO: 23; More preferably, the first antibody comprises a heavy chain variable region as shown in SEQ ID NO: 15 and a light chain variable region as shown in SEQ ID NO: 16, and the second antibody comprises a heavy chain variable region as shown in SEQ ID NO: 29 and a light chain variable region as shown in SEQ ID NO:

30.

5. A protein comprising an antigen-binding domain, said antigen-binding domain comprising the antibody or antigen-binding fragment thereof of any one of claims 1-3 or the antibody composition of claim 4; Preferably, the protein is a chimeric antigen receptor, which includes an antigen-binding domain, a transmembrane domain, and an intracellular signal transduction domain. Preferably, the protein is a multispecific antibody or its antigen-binding fragment, wherein the multispecific antibody or its antigen-binding fragment comprises two or more antigen-binding domains.

6. A biomaterial relating to the antibody or antigen-binding fragment thereof of any one of claims 1-3, the antibody composition of claim 4, or the protein of claim 5, said biomaterial comprising any one of a1)-a9): a1) A nucleic acid molecule encoding the antibody or antigen-binding fragment thereof as described in any one of claims 1-3, the antibody composition of claim 4, or the protein of claim 5; a2) An expression cassette containing the nucleic acid molecule described in a1); a3) A carrier containing the nucleic acid molecule described in a1); a4) A carrier containing the expression box described in a2); a5) Cells containing the nucleic acid molecules described in a1); a6) Cells containing the expression cassette described in a2); a7) Cells containing the carrier described in a3); a8) Cells containing the carrier described in a4); a9) A cell comprising the antibody or antigen-binding fragment thereof as described in any one of claims 1-3, the antibody composition of claim 4, or the protein of claim 5; None of the cells described in a5)-a9) contain reproductive material.

7. An antibody-drug conjugate comprising: an antibody or an antigen-binding fragment thereof according to any one of claims 1-3 or an antibody composition according to claim 4; and a drug covalently linked to said antibody or an antigen-binding fragment thereof.

8. A pharmaceutical composition comprising: an antibody or an antigen-binding fragment thereof according to any one of claims 1-3, an antibody composition according to claim 4, a protein according to claim 5, a biological material according to claim 6, or an antibody-drug conjugate according to claim 7; and a pharmaceutically acceptable carrier.

9. Diagnostic or therapeutic reagent kits, comprising: The antibody or antigen-binding fragment thereof of any one of claims 1-3, the antibody composition of claim 4, the protein of claim 5, the biomaterial of claim 6, the antibody-drug conjugate of claim 7, or the pharmaceutical composition of claim 8.

10. The use of the antibody or antigen-binding fragment thereof according to any one of claims 1-3, the antibody composition according to claim 4, the protein according to claim 5, the biomaterial according to claim 6, the antibody-drug conjugate according to claim 7, or the pharmaceutical composition according to claim 8 in the preparation of a product, wherein the product is used in any one of b1)-b3): b1) Prevention, treatment and / or diagnosis of rabies virus infection or related diseases, b2) Diagnose rabies virus infection or related diseases; b3) Detect rabies virus in the sample, preferably the presence or level of rabies virus G protein; Preferably, the rabies virus infection-related diseases include rabies; Preferably, the sample is whole blood, red blood cell concentrate, platelet concentrate, white blood cell concentrate, tissue, bone marrow aspirate, plasma, serum, cerebrospinal fluid, feces, urine, cultured cells, saliva, oral secretions and / or nasal secretions.

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