Anti-monkeypox virus antibodies or antigen-binding fragments thereof, reagents and kits for detecting monkeypox virus

Abstract

The present invention discloses anti-monkeypox virus antibodies or antigen-binding fragments thereof, reagents and kits for detecting monkeypox virus, and relates to the field of antibodies. The anti-monkeypox virus antibodies disclosed herein include heavy chain complementary determining regions and light chain complementary determining regions. The antibodies provide an important source of raw materials for the detection of monkeypox virus and have improved affinity or activity.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This disclosure claims priority to Chinese patent application number 202211130858.0 filed with the China Patent Office on September 16, 2022, entitled “Anti-monkeypox virus antibodies or functional fragments thereof, reagents and kits for detecting monkeypox virus,” the entire contents of which are incorporated by reference into this disclosure. Technical Field

[0003] The present invention relates to the field of antibody technology, and in particular to anti-monkeypox virus antibodies or antigen-binding fragments thereof, and reagents and kits for detecting monkeypox virus. Background Art

[0004] Monkeypox is caused by the monkeypox virus (MPXV). The monkeypox virus is a double-stranded DNA virus belonging to the genus Orthopoxvirus in the family Poxviridae. Monkeypox is a viral zoonosis that occurs primarily in the tropical rainforests of Central and West Africa and is occasionally exported to other regions. Current studies have shown that the virus invades the human body through mucous membranes and broken skin. Humans are primarily infected through contact with exudates, blood, or other body fluids from infected animal lesions, or through bites or scratches from infected animals. Transmission from person to person is primarily through close contact, but can also be spread through droplets, contact with objects contaminated with the virus, and vertical transmission through the placenta. Sexual transmission cannot be ruled out.

[0005] Clinically, monkeypox symptoms are similar to those of smallpox, but are milder, with an incubation period of 5-12 days, often 6-13 days. Initial symptoms include fever, headache, swollen lymph nodes, muscle aches, and severe fatigue, followed by a rash on the face and body. Monkeypox is a self-limiting disease, typically resolving within two to three weeks. However, in children, pregnant women, or those with immunosuppression due to other health conditions, monkeypox can lead to secondary infections, such as pneumonia, sepsis, and encephalitis, among other serious illnesses.

[0006] Currently, the primary methods for detecting monkeypox virus are PCR amplification and immunoassays. PCR amplification is the predominant method used both domestically and internationally, but it places high demands on instrumentation, testing sites, and environmental conditions. It also suffers from drawbacks such as long testing times and low throughput, making it unsuitable for large-scale population testing. Colloidal gold-based immunoassays, on the other hand, offer advantages such as speed, convenience, and portability, making them a hot topic for research and development. However, the performance of colloidal gold detection relies on the performance of antibodies against monkeypox virus.

[0007] Therefore, there is a strong need in the art for antibodies that can effectively bind to and detect monkeypox virus. Summary of the Invention

[0008] The object of the present invention is to provide anti-monkeypox virus antibodies or antigen-binding fragments thereof, reagents and kits for detecting monkeypox virus or monkeypox virus antigens.

[0009] The present invention is achieved in that:

[0010] In a first aspect, an embodiment of the present invention provides an antibody or an antigen-binding fragment thereof, comprising HCDR1, HCDR2, HCDR3 and LCDR1, LCDR2, LCDR3, wherein the HCDR1 / HCDR2 / HCDR3 combination is the same as the HCDR1 / HCDR2 / HCDR3 combination contained in the heavy chain variable region shown in any one of SEQ ID NO:23 to SEQ ID NO:30, and the LCDR1 / LCDR2 / LCDR3 combination is the same as the LCDR1 / LCDR2 / LCDR3 combination contained in the light chain variable region shown in any one of SEQ ID NO:31 to SEQ ID NO:34.

[0011] In an optional embodiment, the amino acid sequence of the HCDR1 comprises SEQ ID NO: 1, or as shown in SEQ ID NO: 1; the amino acid sequence of the HCDR2 comprises SEQ ID NO: 2 or 3, or as shown in SEQ ID NO: 2 or 3; the amino acid sequence of the HCDR3 comprises SEQ ID NO: 47, or as shown in SEQ ID NO: 47; the amino acid sequence of the LCDR1 comprises SEQ ID NO: 4, or as shown in SEQ ID NO: 4; the amino acid sequence of the LCDR2 comprises SEQ ID NO: 5, 6, 7 or 8, or as shown in SEQ ID NO: 5, 6, 7 or 8; the amino acid sequence of the LCDR3 comprises SEQ ID NO: 9, or as shown in SEQ ID NO: 9.

[0012] In a second aspect, an embodiment of the present invention provides an antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises a sequence structure of HFR1-HCDR1-HFR2-HCDR2-HFR3-HCDR3-HFR4, and the light chain variable region comprises a sequence structure of LFR1-LCDR1-LFR2-LCDR2-LFR3-LCDR3-LFR4, and the amino acid sequences of the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 are the above-mentioned amino acid sequences of HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3.

[0013] In a third aspect, an embodiment of the present invention provides an antibody or an antigen-binding fragment thereof, wherein the binding epitope of the antibody or antigen-binding fragment thereof is the same as the binding epitope of the antibody or antigen-binding fragment thereof described in any one of the above items; or, the antibody or antigen-binding fragment thereof competitively binds to the same epitope as the antibody or antigen-binding fragment thereof described in any one of the above items.

[0014] In a fourth aspect, an embodiment of the present invention provides an antibody conjugate comprising the antibody or antigen-binding fragment thereof as described in the preceding embodiments.

[0015] In a fifth aspect, an embodiment of the present invention provides a reagent or kit, which includes the antibody or antigen-binding fragment thereof as described in the above embodiments or the antibody conjugate as described in the above embodiments.

[0016] In a sixth aspect, an embodiment of the present invention provides a method for detecting monkeypox virus or monkeypox virus antigen, which comprises: contacting the antibody or antigen-binding fragment thereof, antibody conjugate or reagent or kit as described in the above embodiments with the monkeypox virus or monkeypox virus antigen in the sample to be tested to form an immune complex.

[0017] In a seventh aspect, an embodiment of the present invention provides an isolated nucleic acid encoding the antibody or antigen-binding fragment thereof described in the preceding embodiments.

[0018] In an eighth aspect, an embodiment of the present invention provides a vector comprising the isolated nucleic acid described in the preceding embodiment.

[0019] In a ninth aspect, an embodiment of the present invention provides a cell containing the isolated nucleic acid described in the preceding embodiment or the vector described in the preceding embodiment.

[0020] In a tenth aspect, an embodiment of the present invention provides a method for preparing the antibody or antigen-binding fragment thereof described in the preceding embodiment, comprising: culturing the cells described in the preceding embodiment.

[0021] In an eleventh aspect, embodiments of the present invention provide the use of the antibodies or antigen-binding fragments thereof, antibody conjugates, reagents or kits as described in the preceding embodiments for detecting monkeypox virus or monkeypox virus antigens or for preparing products for detecting monkeypox virus or monkeypox virus antigens.

[0022] The present invention has the following beneficial effects:

[0023] The anti-monkeypox virus antibody disclosed in the present invention comprises the heavy chain complementary determining region and the light chain complementary determining region. The antibody provides an important source of raw materials for the detection of monkeypox virus or monkeypox virus antigen and has improved affinity or activity. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for use in the embodiments. It should be understood that the following drawings only illustrate certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other relevant drawings can be obtained based on these drawings without paying any creative work.

[0025] Figure 1 The results of reducing SDS-PAGE of Anti-MPXV 6F13RMb1 to 3 are shown. DETAILED DESCRIPTION

[0026] To make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention are described clearly and completely below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer are used. Where the manufacturer of the reagents or instruments is not specified, all are conventional products that can be purchased commercially.

[0027] The features and performance of the present invention are further described in detail below with reference to the embodiments.

[0028] In a first aspect, an embodiment of the present invention provides an antibody or an antigen-binding fragment thereof, comprising HCDR1, HCDR2, HCDR3 and LCDR1, LCDR2, LCDR3, wherein the HCDR1 / HCDR2 / HCDR3 combination is the same as the HCDR1 / HCDR2 / HCDR3 combination contained in the heavy chain variable region shown in any one of SEQ ID NO:23 to SEQ ID NO:30, and the LCDR1 / LCDR2 / LCDR3 combination is the same as the LCDR1 / LCDR2 / LCDR3 combination contained in the light chain variable region shown in any one of SEQ ID NO:31 to SEQ ID NO:34.

[0029] It should be noted that the HCDR1, HCDR2 and HCDR3 have amino acid sequences consistent with those of HCDR1, HCDR2 and HCDR3 in the same heavy chain variable region, and the LCDR1, LCDR2 and LCDR3 have amino acid sequences consistent with those of LCDR1, LCDR2 and LCDR3 in the same light chain variable region.

[0030] For example, the HCDR1, HCDR2, and HCDR3 have amino acid sequences consistent with those of HCDR1, HCDR2, and HCDR3 in the heavy chain variable region shown in SEQ ID NO: 23; and the LCDR1, LCDR2, and LCDR3 have amino acid sequences consistent with those of LCDR1, LCDR2, and LCDR3 in the light chain variable region shown in SEQ ID NO: 31.

[0031] In the present invention, the term "antibody" is used in the broadest sense and may include full-length monoclonal antibodies, bispecific or multispecific antibodies, and chimeric antibodies, so long as they exhibit the desired biological activity.

[0032] As used herein, the terms "complementarity determining region," "CDR," or "CDRs" refer to the hypervariable regions of the heavy and light chains of immunoglobulins, and include one or more, or even all, of the amino acid residues that contribute substantially to the binding of an antibody or antigen-binding fragment to its recognized antigen or epitope. In specific embodiments of the present invention, CDRs refer to the hypervariable regions of the heavy and light chains of the antibody.

[0033] In the present invention, the heavy chain complementary determining region is represented by HCDR, and the three CDRs contained in the heavy chain variable region include HCDR1, HCDR2 and HCDR3; the light chain complementary determining region is represented by LCDR, and the three CDRs contained in the light chain variable region include LCDR1, LCDR2 and LCDR3.

[0034] In alternative embodiments, the CDRs may be defined according to the Kabat, Chothia, AbM, Contact, or IMGT systems.

[0035] Kabat et al. were the first to propose a standardized numbering scheme for immunoglobulin variable regions. Over the past few decades, the accumulation of sequences has led to the creation of the Kabat database, and the Kabat numbering scheme is generally considered to be the widely adopted standard for numbering antibody residues. There are other definitions of CDRs that may not strictly follow one of the above methods but will still overlap with at least a portion of the Kabat-defined CDRs, although they may be shortened or lengthened based on predictions or experimental results for specific residues or groups of residues. As used herein, CDRs can refer to CDRs defined by any method known in the art, including combinations of methods.

[0036] In alternative embodiments, the CDRs may refer to CDRs defined by any method known in the art.

[0037] In an optional embodiment, the CDRs are based on the CDRs defined by the Kabat system, with the N-terminus of HCDR3 extended by 1 amino acid and the C-terminus shortened by 2 amino acids; the C-terminus of LCDR3 is shortened by 2 amino acids.

[0038] In an optional embodiment, the HCDR1 comprises the amino acid sequence shown in SEQ ID NO: 1; the HCDR2 comprises the amino acid sequence shown in SEQ ID NO: 2 or 3; the HCDR3 comprises the amino acid sequence shown in SEQ ID NO: 47; the LCDR1 comprises the amino acid sequence shown in SEQ ID NO: 4; the LCDR2 comprises the amino acid sequence shown in SEQ ID NO: 5, 6, 7 or 8; and the LCDR3 comprises the amino acid sequence shown in SEQ ID NO: 9.

[0039] In a second aspect, an embodiment of the present invention provides an antibody or an antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment thereof comprises HCDR1, HCDR2, HCDR3 and LCDR1, LCDR2, LCDR3, wherein the HCDR1 comprises the amino acid sequence shown in SEQ ID NO: 1; the HCDR2 comprises the amino acid sequence shown in SEQ ID NO: 2 or 3; the HCDR3 comprises the amino acid sequence shown in SEQ ID NO: 47; the LCDR1 comprises the amino acid sequence shown in SEQ ID NO: 4; the LCDR2 comprises the amino acid sequence shown in SEQ ID NO: 5, 6, 7 or 8; and the LCDR3 comprises the amino acid sequence shown in SEQ ID NO: 9.

[0040] In an optional embodiment, the HCDR1, HCDR2, HCDR3 and LCDR1, LCDR2, LCDR3 correspond to the amino acids in the H31-H35, H50-H65, H94-H96, L24-34, L50-56, L89-95 position segments under Kabat numbering, respectively.

[0041] In an optional embodiment, the amino acid sequence of HCDR1 in the antibody or antigen-binding fragment thereof described in the first or second aspect above is shown as SEQ ID NO: 1; the amino acid sequence of HCDR2 is shown as SEQ ID NO: 2 or 3; the amino acid sequence of HCDR3 is shown as SEQ ID NO: 47; the amino acid sequence of LCDR1 is shown as SEQ ID NO: 4; the amino acid sequence of LCDR2 is shown as SEQ ID NO: 5, 6, 7 or 8; and the amino acid sequence of LCDR3 is shown as SEQ ID NO: 9.

[0042] In an optional embodiment, the amino acid sequence of HCDR1 in the antibody or antigen-binding fragment thereof described in the first or second aspect above is shown as SEQ ID NO: 1; the amino acid sequence of HCDR2 is shown as SEQ ID NO: 2; the amino acid sequence of HCDR3 is shown as SEQ ID NO: 47; the amino acid sequence of LCDR1 is shown as SEQ ID NO: 4; the amino acid sequence of LCDR2 is shown as SEQ ID NO: 5, 6, 7 or 8; and the amino acid sequence of LCDR3 is shown as SEQ ID NO: 9.

[0043] In an optional embodiment, the amino acid sequence of HCDR1 in the antibody or antigen-binding fragment thereof described in the first or second aspect above is shown as SEQ ID NO: 1; the amino acid sequence of HCDR2 is shown as SEQ ID NO: 3; the amino acid sequence of HCDR3 is shown as SEQ ID NO: 47; the amino acid sequence of LCDR1 is shown as SEQ ID NO: 4; the amino acid sequence of LCDR2 is shown as SEQ ID NO: 5, 6, 7 or 8; and the amino acid sequence of LCDR3 is shown as SEQ ID NO: 9.

[0044] In an alternative embodiment, the antibody or antigen-binding fragment thereof further comprises HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3 and LFR4.

[0045] In an alternative embodiment, the HFR1 to HFR4 include, in sequence, SEQ ID NO: 10 to SEQ ID NO: 13, or an amino acid sequence having at least 80% identity thereto.

[0046] In an alternative embodiment, the HFR1 to HFR4 are represented by SEQ ID NO: 10 to SEQ ID NO: 13, or an amino acid sequence having at least 80% identity thereto.

[0047] In an alternative embodiment, the LFR1 to LFR4 include, in sequence, SEQ ID NO: 19 to SEQ ID NO: 22, or an amino acid sequence having at least 80% identity thereto.

[0048] In an alternative embodiment, LFR1 to LFR4 are represented by SEQ ID NO: 19 to SEQ ID NO: 22, or an amino acid sequence having at least 80% identity thereto.

[0049] In alternative embodiments, the framework region amino acid sequence of the antibody or antigen-binding fragment thereof may be at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the above-mentioned framework region (SEQ ID NO: 10 to SEQ ID NO: 13 or SEQ ID NO: 19 to SEQ ID NO: 22).

[0050] In an optional embodiment, the amino acid sequence of HFR1 comprises SEQ ID NO: 14, or is shown as SEQ ID NO: 14.

[0051] In an alternative embodiment, the amino acid sequence of HFR2 comprises SEQ ID NO: 15, 16 or 17, or is shown as SEQ ID NO: 15, 16 or 17.

[0052] In an alternative embodiment, the amino acid sequence of HFR3 comprises SEQ ID NO: 18, or is shown as SEQ ID NO: 18.

[0053] In an alternative embodiment, the amino acid sequence of LFR1 comprises SEQ ID NO:43, or is shown as SEQ ID NO:43.

[0054] In an alternative embodiment, the amino acid sequence of LFR3 comprises SEQ ID NO: 44, 45 or 46, or is shown in SEQ ID NO: 44, 45 or 46.

[0055] In a third aspect, an embodiment of the present invention provides an antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises a sequence structure of HFR1-HCDR1-HFR2-HCDR2-HFR3-HCDR3-HFR4, and the light chain variable region comprises a sequence structure of LFR1-LCDR1-LFR2-LCDR2-LFR3-LCDR3-LFR4, and the amino acid sequences of the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 are the above-mentioned amino acid sequences of HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3.

[0056] In an optional embodiment, the amino acid sequence of HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3, and LFR4 is the above-mentioned amino acid sequence of HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3, and LFR4.

[0057] In the present invention, the heavy chain variable region is obtained by arranging and connecting the following numbered CDRs and FRs in the following combinations: HFR1-HCDR1-HFR2-HCDR2-HFR3-HCDR3-HFR4; the light chain variable region is obtained by arranging and connecting the following numbered CDRs and FRs in the following combinations: LFR1-LCDR1-LFR2-LCDR2-LFR3-LCDR3-LFR4.

[0058] In an alternative embodiment, the heavy chain variable region comprises an amino acid sequence as shown in any one of SEQ ID NOs: 23 to 30, or an amino acid sequence having at least 80% identity thereto.

[0059] In an alternative embodiment, the heavy chain variable region consists of an amino acid sequence shown in any one of SEQ ID NOs: 23 to 30, or an amino acid sequence having at least 80% identity thereto.

[0060] In an alternative embodiment, the light chain variable region comprises an amino acid sequence as shown in any one of SEQ ID NOs: 31 to 34 or an amino acid sequence having at least 80% identity thereto.

[0061] In an alternative embodiment, the light chain variable region consists of an amino acid sequence shown in any one of SEQ ID NOs: 31 to 34, or an amino acid sequence having at least 80% identity thereto.

[0062] In alternative embodiments, the variable region sequences of the antibodies or antigen-binding fragments thereof may be at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the above variable regions (SEQ ID NOs: 23 to 30 or SEQ ID NOs: 31 to 34).

[0063] In an alternative embodiment, the variant amino acid positions of identity are not in the CDRs region.

[0064] In an optional embodiment, the heavy chain variable region consists of the amino acid sequence shown in any one of SEQ ID NOs: 23 to 30; and the light chain variable region consists of the amino acid sequence shown in any one of SEQ ID NOs: 31 to 34.

[0065] In a fourth aspect, an embodiment of the present invention provides an antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region is composed of the amino acid sequence shown in any one of SEQ ID NOs: 23 to 30; and the light chain variable region is composed of the amino acid sequence shown in any one of SEQ ID NOs: 31 to 34.

[0066] In an optional embodiment, the heavy chain variable region of the antibody or antigen-binding fragment thereof described in the third aspect or the fourth aspect above is composed of the amino acid sequence shown in any one of SEQ ID NOs: 23 to 30; and the light chain variable region is composed of the amino acid sequence shown in SEQ ID NO: 31.

[0067] In an optional embodiment, the heavy chain variable region of the antibody or antigen-binding fragment thereof described in the third aspect or the fourth aspect above is composed of the amino acid sequence shown in any one of SEQ ID NOs: 23 to 30; and the light chain variable region is composed of the amino acid sequence shown in SEQ ID NO: 32.

[0068] In an optional embodiment, the heavy chain variable region of the antibody or antigen-binding fragment thereof described in the third aspect or the fourth aspect above is composed of the amino acid sequence shown in any one of SEQ ID NOs: 23 to 30; and the light chain variable region is composed of the amino acid sequence shown in SEQ ID NO: 33.

[0069] In an optional embodiment, the heavy chain variable region of the antibody or antigen-binding fragment thereof described in the third aspect or the fourth aspect above is composed of the amino acid sequence shown in any one of SEQ ID NOs: 23 to 30; and the light chain variable region is composed of the amino acid sequence shown in SEQ ID NO: 34.

[0070] In an alternative embodiment, the antibody or antigen-binding fragment thereof further comprises a constant region.

[0071] In alternative embodiments, the constant region comprises a heavy chain constant region and / or a light chain constant region.

[0072] In an optional embodiment, the heavy chain constant region is selected from the heavy chain constant region of any one of IgG1, IgG2, IgG3, IgG4, IgA, IgM, IgE and IgD; and the light chain constant region is selected from the κ or λ type light chain constant region.

[0073] In an optional embodiment, the species origin of the constant region is any one of cow, horse, pig, sheep, goat, rat, mouse, dog, cat, rabbit, donkey, deer, mink, chicken, duck, goose and human.

[0074] In an alternative embodiment, the heavy chain constant region comprises an amino acid sequence as shown in SEQ ID NO: 35 or at least 80% identical thereto.

[0075] In an alternative embodiment, the heavy chain constant region consists of an amino acid sequence as shown in SEQ ID NO: 35 or an amino acid sequence at least 80% identical thereto.

[0076] In an alternative embodiment, the light chain constant region comprises an amino acid sequence as shown in SEQ ID NO: 36 or an amino acid sequence at least 80% identical thereto.

[0077] In an alternative embodiment, the light chain constant region consists of an amino acid sequence as shown in SEQ ID NO: 36 or an amino acid sequence at least 80% identical thereto.

[0078] Specifically, the constant region sequence can be at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the above-mentioned constant region (SEQ ID NO: 35 or 36).

[0079] In an alternative embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain and a light chain.

[0080] In an alternative embodiment, the heavy chain comprises an amino acid sequence as shown in any one of SEQ ID NOs: 37 to 39, or an amino acid sequence at least 80% identical thereto.

[0081] In an alternative embodiment, the heavy chain consists of an amino acid sequence as shown in any one of SEQ ID NOs: 37 to 39, or an amino acid sequence at least 80% identical thereto.

[0082] In an alternative embodiment, the light chain comprises an amino acid sequence as shown in any one of SEQ ID NOs: 40 to 42, or an amino acid sequence at least 80% identical thereto.

[0083] In an alternative embodiment, the light chain consists of an amino acid sequence as shown in any one of SEQ ID NOs: 40 to 42, or an amino acid sequence at least 80% identical thereto.

[0084] In alternative embodiments, the heavy or light chain sequence of the antibody or antigen-binding fragment thereof may be at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the above-mentioned heavy or light chain sequences (SEQ ID NOs: 37 to 39 or SEQ ID NOs: 40 to 42).

[0085] In an alternative embodiment, the variant amino acid positions of identity are not in the CDRs region.

[0086] In an optional embodiment, the heavy chain consists of the amino acid sequence shown in any one of SEQ ID NOs: 37 to 39; and the light chain consists of the amino acid sequence shown in any one of SEQ ID NOs: 40 to 42.

[0087] On the other hand, an embodiment of the present invention provides an antibody or an antigen-binding fragment thereof, wherein the binding epitope of the antibody or antigen-binding fragment thereof is the same as the binding epitope of the antibody or antigen-binding fragment thereof described in any one of the above items; or, the antibody or antigen-binding fragment thereof competitively binds to the same epitope as the antibody or antigen-binding fragment thereof described in any one of the above items.

[0088] In an optional embodiment, the same epitope is located within the amino acid sequence 1 to 110 of the monkeypox virus A29L protein; or, the antibody or antigen-binding fragment thereof competes with the antibody or antigen-binding fragment thereof described in any of the above embodiments for binding to the monkeypox virus A29L protein.

[0089] In an optional embodiment, the antibody or antigen-binding fragment thereof binds to a unique epitope of monkeypox virus A29L protein (MKPVA29L), but does not bind to a common epitope of monkeypox virus A29L protein, cowpox virus 162 protein (CPXV162) and vaccinia virus A27L protein (VACA27L).

[0090] In an alternative embodiment, the antibody or antigen-binding fragment thereof binds to an amino acid region between amino acids 17 to 49 of the monkeypox virus A29L protein amino acid sequence.

[0091] In an optional embodiment, the antibody or antigen-binding fragment thereof specifically binds to monkeypox virus or monkeypox virus A29L protein (MKPVA29L) and has no cross-reactivity with cowpox virus 162 protein (CPXV162) or vaccinia virus A27L protein (VACA27L).

[0092] In an alternative embodiment, the antibody or antigen-binding fragment thereof has a KD ≤ 7.91×10 -8Affinity binding of M to monkeypox virus antigens.

[0093] In an alternative embodiment, the antibody or antigen-binding fragment thereof has a KD ≤ 10 -08 M, KD≤10 -09 M, KD≤10 -10 M, KD≤10 -11 M or KD≤10 -12 Affinity binding of M to monkeypox virus antigens.

[0094] In an alternative embodiment, the antibody or antigen-binding fragment thereof has a KD ≤ 1.47×10 -09 M or KD≤9.96×10 -10 Affinity binding of M to monkeypox virus antigens.

[0095] In an optional embodiment, the antigen-binding fragment is selected from any one of F(ab')2, Fab', Fab, Fv and scFv of the antibody.

[0096] Antigen-binding fragments of the above antibodies generally have the same binding specificity as the antibody from which they are derived.

[0097] In the present invention, the terms "antigen-binding fragment" and "antigen-binding fragment" having the same binding specificity as the antibody from which it is derived are used interchangeably.

[0098] Those skilled in the art will readily appreciate, based on the disclosure herein, that antigen-binding fragments of the aforementioned antibodies can be obtained by, for example, enzymatic digestion (including pepsin or papain) and / or chemical reduction to cleave disulfide bonds. Based on the structure of the intact antibody disclosed herein, those skilled in the art can readily obtain the aforementioned antigen-binding fragments.

[0099] The antigen-binding fragments of the above antibodies can also be synthesized by recombinant genetic techniques known to those skilled in the art or by, for example, an automatic peptide synthesizer, such as those sold by Applied BioSystems.

[0100] On the other hand, an embodiment of the present invention further provides an antibody conjugate, which includes the antibody or antigen-binding fragment thereof as described in the above embodiments.

[0101] In an optional embodiment, the antibody conjugate further comprises biotin or a biotin derivative conjugated to the antibody or antigen-binding fragment thereof.

[0102] In an optional embodiment, the antibody conjugate further comprises a label coupled to the antibody or antigen-binding fragment thereof.

[0103] In an optional embodiment, the above-mentioned marker refers to a class of substances with properties that can be directly observed by the naked eye or detected or detected by an instrument, such as luminescence, color development, radioactivity, etc., through which qualitative or quantitative detection of the corresponding target can be achieved.

[0104] In an optional embodiment, the label is selected from at least one of fluorescent dyes, enzymes, radioisotopes, chemiluminescent reagents and nanoparticle labels.

[0105] In actual use, those skilled in the art can select a suitable marker according to the detection conditions or actual needs. No matter which marker is used, it falls within the scope of protection of the present invention.

[0106] In an optional embodiment, the fluorescent dyes include but are not limited to fluorescein dyes and their derivatives (for example, including but not limited to fluorescein isothiocyanate (FITC), hydroxyfluorescein (FAM), tetrachlorofluorescein (TET), etc. or their analogs), rhodamine dyes and their derivatives (for example, including but not limited to red rhodamine (RBITC), tetramethylrhodamine (TAMRA), rhodamine B (TRITC), etc. or their analogs), Cy series dyes and their derivatives (for example, including but not limited to Cy2, Cy3, Cy3B, Cy3.5, C y5, Cy5.5, Cy3, etc. or their analogs), Alexa series dyes and their derivatives (for example, including but not limited to AlexaFluor350, 405, 430, 488, 532, 546, 555, 568, 594, 610, 33, 647, 680, 700, 750, etc. or their analogs) and protein dyes and their derivatives (for example, including but not limited to phycoerythrin (PE), phycocyanin (PC), allophycocyanin (APC), peridinin-chlorophyll protein (preCP), etc.).

[0107] In alternative embodiments, the enzyme includes, but is not limited to, horseradish peroxidase, alkaline phosphatase, β-galactosidase, glucose oxidase, carbonic anhydrase, acetylcholinesterase, and glucose-6-phosphate deoxidase.

[0108] In an optional embodiment, the radioactive isotopes include but are not limited to 212Bi, 131I, 111In, 90Y, 186Re, 211At, 125I, 188Re, 153Sm, 213Bi, 32P, 94mTc, 99mTc, 203Pb, 67Ga, 68Ga, 43Sc, 47Sc, 110mIn, 97Ru, 62Cu, 64Cu, 67Cu, 68Cu, 86Y, 88Y, 121Sn, 161Tb, 166Ho, 105Rh, 177Lu, 172Lu and 18F.

[0109] In an optional embodiment, the chemiluminescent reagent includes but is not limited to luminol and its derivatives, lucigenin, crustacean fluorescein and its derivatives, ruthenium bipyridine and its derivatives, acridinium esters and their derivatives, dioxetanes and their derivatives, lophanes and their derivatives, and peroxalates and their derivatives.

[0110] In an optional embodiment, the nanoparticle markers include but are not limited to nanoparticles, colloids, organic nanoparticles, magnetic nanoparticles, quantum dot nanoparticles and rare earth complex nanoparticles.

[0111] In alternative embodiments, the colloid includes, but is not limited to, colloidal metals, colloidal selenium, disperse dyes, dye-labeled microspheres, and latex.

[0112] In an optional embodiment, the colloidal metal includes but is not limited to colloidal gold or colloidal silver.

[0113] In an optional embodiment, the colloidal metal is colloidal gold.

[0114] In an optional embodiment, the antibody conjugate further comprises a solid phase support coupled to the antibody or antigen-binding fragment thereof. In the antibody conjugate, the antibody is coupled to the solid phase support.

[0115] In an alternative embodiment, the solid support is selected from microspheres, plates and membranes.

[0116] In an optional embodiment, the solid phase includes but is not limited to magnetic microspheres, plastic microspheres, plastic microparticles, microplates, glass, capillaries, nylon and nitrocellulose membranes.

[0117] In an optional embodiment, the solid phase carrier is a nitrocellulose membrane.

[0118] On the other hand, an embodiment of the present invention further provides a reagent or kit, which includes the antibody or antigen-binding fragment thereof as described in any of the foregoing embodiments or the antibody conjugate as described in any of the foregoing embodiments.

[0119] In an alternative embodiment, the reagent or kit has improved detection sensitivity or specificity.

[0120] On the other hand, an embodiment of the present invention further provides a method for detecting monkeypox virus or monkeypox virus antigen, comprising:

[0121] The antibody or antigen-binding fragment thereof as described in any of the preceding embodiments, the antibody conjugate as described in any of the preceding embodiments, or the reagent or kit as described in any of the preceding embodiments is contacted with the monkeypox virus or monkeypox virus antigen in the sample to be tested to form an immune complex.

[0122] In a preferred embodiment, the immune complex further comprises a second antibody that binds to the antibody or antigen-binding fragment thereof.

[0123] In a preferred embodiment, the immune complex further comprises a second antibody, which binds to the monkeypox virus or a monkeypox virus antigen.

[0124] On the other hand, an embodiment of the present invention also provides the use of the antibody or antigen-binding fragment described in any of the preceding embodiments, or the antibody conjugate described in any of the preceding embodiments, or the reagent or kit described in any of the preceding embodiments in detecting monkeypox virus or monkeypox virus antigen or preparing a product for detecting monkeypox virus or monkeypox virus antigen.

[0125] On the other hand, an embodiment of the present invention also provides the use of the antibody or antigen-binding fragment described in any of the preceding embodiments, or the antibody conjugate described in any of the preceding embodiments, or the reagent or kit described in any of the preceding embodiments in the preparation of a product having at least one of the following uses, wherein the uses include: diagnosing or assisting in the diagnosis of diseases related to monkeypox virus infection, and predicting or assisting in the prediction of the prognosis and efficacy of diseases related to monkeypox virus infection.

[0126] In an optional embodiment, the related diseases caused by the monkeypox virus infection include at least one of fever, headache, swollen lymph nodes, muscle aches, severe fatigue, pneumonia, sepsis and encephalitis.

[0127] In an alternative embodiment, the product comprises a reagent or a kit.

[0128] In an alternative embodiment, the reagent or kit has improved detection sensitivity or specificity.

[0129] On the other hand, an embodiment of the present invention further provides an isolated nucleic acid encoding the antibody or antigen-binding fragment thereof according to any of the aforementioned embodiments.

[0130] On the other hand, an embodiment of the present invention further provides a vector comprising the isolated nucleic acid described in any of the aforementioned embodiments.

[0131] On the other hand, an embodiment of the present invention further provides a cell comprising the isolated nucleic acid described in any of the foregoing embodiments or the vector described in any of the foregoing embodiments.

[0132] On the other hand, an embodiment of the present invention further provides a method for preparing the antibody or antigen-binding fragment thereof as described in any of the preceding embodiments, comprising: culturing the cell as described in any of the preceding embodiments.

[0133] Based on the amino acid sequence of the antibody or antigen-binding fragment thereof disclosed in the present invention, those skilled in the art can easily conceive of using genetic engineering technology or other technologies (chemical synthesis, recombinant expression) to prepare the antibody or antigen-binding fragment thereof. For example, the antibody or antigen-binding fragment thereof can be isolated and purified from the culture product of recombinant cells that can recombinantly express the antibody or antigen-binding fragment thereof as described in any of the above items. This is easy to achieve for those skilled in the art. Based on this, regardless of the technology used to prepare the antibody or antigen-binding fragment of the present invention, it falls within the scope of protection of the present invention.

[0134] To make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention are described clearly and completely below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer are used. Where the manufacturer of the reagents or instruments is not specified, all are conventional products that can be purchased commercially.

[0135] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art to which this disclosure belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the preparations or unit doses herein, some methods and materials are now described. Unless otherwise stated, the techniques employed or contemplated herein are standard methods. Materials, methods, and examples are illustrative and non-limiting only.

[0136] The practice of the present invention will employ, unless otherwise indicated, conventional techniques of cell biology, molecular biology (including recombinant techniques), microbiology, biochemistry, and immunology, which are within the capabilities of a person skilled in the art. The technique is fully explained in the literature, for example, in Molecular Cloning: A Laboratory Manual, 2nd ed. (Sambrook et al., 1989); Oligonucleotide Synthesis (MJ Gait, ed., 1984); Animal Cell Culture (RI Freshney, ed., 1987); Methods in Enzymology (Academic Press, Inc.); Handbook of Experimental Immunology (DM Weir and CC Blackwell, eds.); Gene Transfer Vectors for Mammalian Cells (JM Miller and MP Calos, eds., 1987); Current Protocols in Molecular Biology (FM Ausubel et al., eds., 1987); and PCR: The Polymerase Chain Reaction. Reaction" (Mullis et al., eds., 1994); and Current Protocols in Immunology (JE Coligan et al., eds., 1991), each of which is expressly incorporated herein by reference.

[0137] The features and performance of the present invention are further described in detail below with reference to the embodiments.

[0138] Example 1 Preparation of Anti-MPXV 6F13 Monoclonal Antibody

[0139] In this example, restriction endonucleases and Prime Star DNA polymerase were purchased from Takara. MagExtractor-RNA extraction kit was purchased from TOYOBO. BD SMART TMThe RACE cDNA Amplification Kit was purchased from Takara. The pMD-18T vector was also purchased from Takara. The plasmid extraction kit was purchased from Tiangen. Primer synthesis and gene sequencing were performed by a gene sequencing company. The hybridoma cell line secreting the anti-MPXV 6F13 monoclonal antibody was generated in our laboratory and revived for future use.

[0140] 1. Expression Plasmid Construction

[0141] In this example, restriction endonucleases and Prime Star DNA polymerase were purchased from Takara. MagExtractor-RNA extraction kit was purchased from TOYOBO. BD SMART TM The RACE cDNA Amplification Kit was purchased from Takara. The pMD-18T vector was also purchased from Takara. The plasmid extraction kit was purchased from Tiangen. Primer synthesis and gene sequencing were performed by a gene sequencing company.

[0142] 1.1 Anti-MPXV 6F13 antibody gene preparation

[0143] mRNA was extracted from a hybridoma cell line secreting the Anti-MPXV 6F13 monoclonal antibody. DNA was obtained by RT-PCR and inserted into the pMD-18T vector. The DNA was then transformed into DH5α competent cells. After colonies were grown, four positive clones each for the Heavy Chain and Light Chain genes were collected and sent to a gene sequencing company for sequencing.

[0144] 1.2 Sequence Analysis of the Anti-MPXV 6F13 Antibody Variable Region Gene

[0145] The gene sequences obtained by the above sequencing were placed in the Kabat antibody database for analysis, and VNTI11.5 software was used for analysis to determine that the genes amplified by the heavy chain and light chain primer pairs were correct. Among them, in the gene fragment amplified by the Light Chain, the VL gene sequence was 336 bp, preceded by a 57 bp leader peptide sequence; in the gene fragment amplified by the Heavy Chain primer pair, the VH gene sequence was 339 bp, belonging to the VH1 gene family, and preceded by a 57 bp leader peptide sequence.

[0146] 1.3 Construction of recombinant antibody expression plasmid

[0147] pcDNA TM 3.4 Vector is a constructed recombinant antibody eukaryotic expression vector, which has been modified to introduce multiple cloning restriction enzyme sites and is subsequently referred to as 3.4A expression vector. Based on the sequencing results of the antibody variable region genes in the above-mentioned pMD-18T, VL and VH gene-specific primers of the Anti-MPXV 6F13 antibody were designed, with restriction endonuclease sites and protective bases at both ends, respectively. PCR amplification was used to amplify the 0.74KB Light Chain gene fragment and the 1.41kb Heavy Chain gene fragment.

[0148] The Heavy Chain and Light Chain gene fragments were double-digested with restriction endonucleases, and the 3.4A vector was double-digested with restriction endonucleases. After the fragments and vectors were purified and recovered, the Heavy Chain gene and Light Chain gene were respectively connected to the 3.4A expression vector to obtain the recombinant expression plasmids of Heavy Chain and Light Chain, respectively.

[0149] 2. Recombinant Antibody Sample Preparation

[0150] Resuscitate HEK293 cells in advance and subculture them into 200 ml system to make the cell density reach 3-5×10 6 cells / ml, cell viability>95%; wash the cells by centrifugation, resolubilize with culture medium, and adjust the cell density to 2.9×10 6 cells / ml, as the cell dilution. Use culture medium to prepare plasmid DNA and transfection reagent dilution, respectively. Add the transfection reagent dilution to the plasmid DNA dilution, mix well, and let it stand at room temperature for 15 minutes; slowly add the mixture to the cell dilution within 1 minute, mix well, take samples and count, record and observe the viability of the cells after transfection, and place them in a 35°C constant temperature incubator for culture, speed of 120 rpm, CO2 content of 8%, and centrifuge after 13 days. The centrifuged supernatant is affinity purified using a protein A affinity chromatography column. Take 6μg of the purified antibody for reducing SDS-PAGE, and the electrophoresis is shown in the figure. After reducing SDS-PAGE, two bands are shown, one with Mr of 50KD (heavy chain) and the other with Mr of 28KD (light chain).

[0151] 3. Affinity and activity optimization

[0152] Although the Anti-MPXV 6F13 monoclonal antibody prepared above has the ability to bind to monkeypox virus antigens, its affinity and antibody activity are not ideal. Therefore, the applicant conducted targeted mutagenesis on the light chain CDR and heavy chain CDR of the antibody. That is, computer simulation of the antibody variable region structure, the structure of the complex interaction between the antigen and the antibody variable region, analysis of the key amino acids of the antibody, and mutation design were performed. According to the mutation plan, bidirectional primers covering the mutation site were designed and synthesized. Primers at both ends of the target DNA were synthesized, and a high-fidelity PCR reaction was performed. The PCR product was cloned into a vector, and the mutant antibody was prepared according to the method of step 2 above. After screening, monoclonal antibodies with significantly improved affinity and antibody activity were obtained and named Anti-MPXV 6F13RMb1 to Anti-MPXV 6F13RMb3. The amino acid sequences of their heavy and light chains are as follows.

[0153] Table 1 Antibody sequences

[0154] Sample name Heavy chain sequence number Light chain number Anti-MPXV 6F13RMb1 SEQ ID NO:37 SEQ ID NO:40 Anti-MPXV 6F13RMb2 SEQ ID NO:38 SEQ ID NO:41 Anti-MPXV 6F13RMb3 SEQ ID NO:39 SEQ ID NO:42

[0155] Example 2 Affinity Analysis

[0156] Purified antibodies were diluted in advance, and recombinant monkeypox antigen (purchased from Feipeng Bio) was serially diluted. Binding and dissociation curves were measured on a Biacore 8K+ instrument using a CM5 chip pre-coupled with goat anti-mouse IgG. The instrument automatically fitted the affinity constant, association rate, and dissociation rate. (KD represents the equilibrium dissociation constant (affinity constant); ka represents the association rate; and kd represents the dissociation rate.)

[0157] Table 2 Affinity analysis data

[0158] Sample name KD(M) ka kd Control Antibody 7.91E-08 1.08E+06 8.55E-02 Anti-MPXV 6F13RMb1 9.96E-10 1.64E+06 1.63E-03 Anti-MPXV 6F13RMb2 1.47E-09 1.09E+06 1.60E-03 Anti-MPXV 6F13RMb3 1.17E-09 2.75E+06 3.23E-03

[0159] Example 3 Activity Identification

[0160] The coating solution (main component NaHCO3) was diluted with recombinant monkeypox recombinant antigen (purchased from Feipeng Bio) to 1ug / ml, 100uL per well, and incubated at 4°C overnight; the next day, the cells were washed twice with washing solution (main component Na2HPO4+Nacl) and patted dry; blocking solution (20% BSA+80% PBS) was added, 120uL per well, incubated at 37°C for 1h, and patted dry; diluted purified antibody and control antibody were added, 100uL / well, incubated at 37°C for 30min; the cells were washed 5 times with washing solution and patted dry; goat anti-mouse IgG-HRP was added, 100uL per well, incubated at 37°C for 30min; the cells were washed 5 times with washing solution and patted dry; colorimetric solution A (50uL / well) and colorimetric solution B (50uL / well) were added for 10min; stop solution was added, 50uL / well; OD value was read at 450nm (reference 630nm) on a microplate reader.

[0161] Table 3 Activity data

[0162] Concentration (ng / ml) 15.63 7.81 3.91 1.95 0.98 0.00 Control Antibody 1.123 0.576 0.307 0.162 0.081 0.023 Anti-MPXV6F13RMb1 1.756 1.117 0.548 0.207 0.108 0.013 Anti-MPXV6F13RMb2 1.972 1.361 0.742 0.437 0.274 0.030 Anti-MPXV6F13RMb3 2.016 1.452 0.729 0.333 0.131 0.026

[0163] Example 4 Antibody Stability Assessment

[0164] The above-mentioned antibodies were placed at 4°C (refrigerator), -80°C (freezer), and 37°C (incubator) for 21 days. Samples were collected at 7, 14, and 21 days for status observation, and the 21-day sample was tested for activity. The results showed that no significant changes in protein status were observed under all three test conditions for 21 days, and the activity did not decrease with increasing test temperature, indicating that the expressed antibodies were stable. The table below shows the OD results of the enzyme immunoassay activity assay of Anti-MPXV 6F13RMb1 after 21 days of testing.

[0165] Table 4 Stability data

[0166] Sample concentration (ng / ml) 15.63 7.81 0.00 4℃, 21-day samples 1.723 1.134 0.012 -80℃, 21-day sample 1.772 1.126 0.011 37℃, 21-day samples 1.736 1.118 0.011

[0167] Example 5 Identification of Antibody Cross-Reactivity

[0168] The cross-reactivity of the antibody prepared above with monkeypox virus A29L protein, cowpox virus 162 protein, and vaccinia virus A27L protein was determined by ELISA. The ELISA procedure was as follows:

[0169] Coating: Coat a microtiter plate with 0.5 μg / ml and 0.05 μg / ml of MKPVA29L, VCA27L, and CPXV162 at 100 μl / well, respectively, and incubate at 37°C for 2 h.

[0170] Blocking: Take 120ul of BSA blocking solution and incubate at 37℃ for 2h.

[0171] Primary antibody / sample: The above antibodies were diluted to 0.5ug / ml and 0.05ug / ml with PBS, 50ul / well, and incubated at 37℃ for 30min.

[0172] Negative control: add PBS and incubate

[0173] Secondary antibody: 100ul / well, add goat anti-mouse IgG-HRP, incubate at 37℃ for 30min.

[0174] Color development: add 50ul of solution A and B respectively, add 50ul of stop solution after 10 minutes, and read the results.

[0175] The results are shown in the following table:

[0176] Table 5 Cross-reactivity data

[0177]

[0178]

[0179] The foregoing description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Those skilled in the art will readily appreciate that various modifications and variations of the present invention are possible. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the present invention are intended to be within the scope of protection of the present invention.

[0180] Some of the amino acid sequences involved in this application are shown in Table 6 below:

[0181] Table 6 Amino acid sequence

[0182]

[0183]

[0184]

Claims

1. An anti-monkeypox virus antibody or antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment thereof comprises HCDR1, HCDR2, HCDR3 and LCDR1, LCDR2, LCDR3, characterized in that: The HCDR1, HCDR2, and HCDR3 combination is the same as the HCDR1, HCDR2, and HCDR3 combination contained in the heavy chain variable region of SEQ ID NO: 23, and the LCDR1, LCDR2, and LCDR3 combination is the same as the LCDR1, LCDR2, and LCDR3 combination contained in the light chain variable region of SEQ ID NO: 31; or; The HCDR1, HCDR2, and HCDR3 combination is the same as the HCDR1, HCDR2, and HCDR3 combination contained in the heavy chain variable region of SEQ ID NO: 24, and the LCDR1, LCDR2, and LCDR3 combination is the same as the LCDR1, LCDR2, and LCDR3 combination contained in the light chain variable region of SEQ ID NO: 32; or; The HCDR1, HCDR2, and HCDR3 combination is the same as the HCDR1, HCDR2, and HCDR3 combination contained in the heavy chain variable region set forth in SEQ ID NO: 25, and the LCDR1, LCDR2, and LCDR3 combination is the same as the LCDR1, LCDR2, and LCDR3 combination contained in the light chain variable region set forth in SEQ ID NO: 33; The CDRs are defined by the Kabat, Chothia, AbM, Contact, or IMGT systems.

2. An anti-monkeypox virus antibody or antigen-binding fragment thereof, comprising HCDR1, HCDR2, HCDR3 and LCDR1, LCDR2, LCDR3, characterized in that: The amino acid sequence of HCDR1 is shown in SEQ ID NO: 1, the amino acid sequence of HCDR2 is shown in SEQ ID NO: 3, the amino acid sequence of HCDR3 is shown in SEQ ID NO: 47, the amino acid sequence of LCDR1 is shown in SEQ ID NO: 4, the amino acid sequence of LCDR2 is shown in SEQ ID NO: 5, and the amino acid sequence of LCDR3 is shown in SEQ ID NO: 9; or The amino acid sequence of HCDR1 is shown in SEQ ID NO: 1, the amino acid sequence of HCDR2 is shown in SEQ ID NO: 2, the amino acid sequence of HCDR3 is shown in SEQ ID NO: 47, the amino acid sequence of LCDR1 is shown in SEQ ID NO: 4, the amino acid sequence of LCDR2 is shown in SEQ ID NO: 6, and the amino acid sequence of LCDR3 is shown in SEQ ID NO: 9; or The amino acid sequence of HCDR1 is shown in SEQ ID NO: 1, the amino acid sequence of HCDR2 is shown in SEQ ID NO: 2, the amino acid sequence of HCDR3 is shown in SEQ ID NO: 47, the amino acid sequence of LCDR1 is shown in SEQ ID NO: 4, the amino acid sequence of LCDR2 is shown in SEQ ID NO: 7, and the amino acid sequence of LCDR3 is shown in SEQ ID NO:

9.

3. The antibody or antigen-binding fragment thereof according to claim 1 or 2, characterized in that The antibody or antigen-binding fragment thereof further includes HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3 and LFR4; the HFR1 to HFR4 are respectively represented by SEQ ID NO:10 to SEQ ID NO:13 or an amino acid sequence having at least 80% identity thereto; the LFR1 to LFR4 are respectively represented by SEQ ID NO:19 to SEQ ID NO:22 or an amino acid sequence having at least 80% identity thereto.

4. An anti-monkeypox virus antibody or antigen-binding fragment thereof, comprising a heavy chain variable region and a light chain variable region, characterized in that: The heavy chain variable region and the light chain variable region are any combination of the following composition: 。 5. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 2 and 4, wherein: The antibody or antigen-binding fragment thereof further comprises a constant region.

6. The antibody or antigen-binding fragment thereof according to claim 5, characterized in that The constant region includes a heavy chain constant region and / or a light chain constant region.

7. The antibody or antigen-binding fragment thereof according to claim 6, characterized in that The heavy chain constant region is selected from the heavy chain constant region of any one of IgG1, IgG2, IgG3, IgG4, IgA, IgM, IgE and IgD; and / or the light chain constant region is selected from the κ type or λ type light chain constant region.

8. The antibody or antigen-binding fragment thereof according to claim 5, characterized in that The species origin of the constant region is any one of cattle, horses, pigs, sheep, goats, rats, mice, dogs, cats, rabbits, donkeys, deer, minks, chickens, ducks, geese and humans.

9. The antibody or antigen-binding fragment thereof according to claim 6, wherein The heavy chain constant region consists of an amino acid sequence shown in SEQ ID NO: 35 or a sequence having at least 80% identity thereto.

10. The antibody or antigen-binding fragment thereof according to claim 6, wherein The light chain constant region consists of an amino acid sequence shown in SEQ ID NO: 36 or a sequence having at least 80% identity thereto.

11. An anti-monkeypox virus antibody or antigen-binding fragment thereof, comprising a heavy chain and a light chain, characterized in that: The heavy chain and light chain are any combination of the following composition: 。 12. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 2, 4, 6 to 11, wherein: The antibody or antigen-binding fragment thereof has a KD of ≤7.91×10 -8 Affinity binding of M to monkeypox virus antigens.

13. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 2 and 4, wherein: The antigen-binding fragment is selected from any one of F(ab')2, Fab', Fab, Fv and scFv of the antibody.

14. An antibody conjugate, characterized in that It comprises the antibody or antigen-binding fragment thereof according to any one of claims 1 to 13.

15. The antibody conjugate according to claim 14, characterized in that The antibody conjugate includes biotin conjugated to the antibody or antigen-binding fragment thereof.

16. The antibody conjugate according to claim 14, characterized in that The antibody conjugate comprises a solid phase carrier coupled to the antibody or antigen-binding fragment thereof.

17. The antibody conjugate according to claim 14, characterized in that The antibody conjugate includes a label coupled to the antibody or antigen-binding fragment thereof.

18. The antibody conjugate according to claim 17, characterized in that The label is selected from at least one of fluorescent dyes, enzymes, radioisotopes, chemiluminescent reagents and nanoparticle labels.

19. The antibody conjugate according to claim 17, characterized in that The label is colloidal gold.

20. A reagent or kit, characterized in that It includes the antibody or antigen-binding fragment thereof according to any one of claims 1 to 13 or the antibody conjugate according to any one of claims 14 to 19.

21. Use of the antibody or antigen-binding fragment thereof according to any one of claims 1 to 13, or the antibody conjugate according to any one of claims 14 to 19, in the preparation of a reagent or kit for detecting monkeypox virus or monkeypox virus antigens.

22. An isolated nucleic acid, characterized in that It encodes the antibody or antigen-binding fragment thereof according to any one of claims 1 to 13.

23. A carrier, characterized in that It contains the nucleic acid according to claim 22.

24. A cell, characterized in that It contains the nucleic acid according to claim 22 or the vector according to claim 23.

25. A method for preparing the antibody or antigen-binding fragment thereof according to any one of claims 1 to 13, characterized in that: It includes: Cultivate the cell of claim 24.

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