Anti-adenovirus antibodies, reagents and kits for detecting adenovirus

Abstract

The present invention discloses an anti-adenovirus antibody, a reagent and a kit for detecting adenovirus, and relates to the field of antibody technology. The anti-adenovirus antibody disclosed in the present invention comprises a heavy chain complementary determining region and a light chain complementary determining region. The antibody provides an important source of raw materials for the detection of adenovirus and has improved activity.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] The present disclosure claims priority to a Chinese patent application with application number 202210684540.0 filed with the Chinese Patent Office on June 17, 2022, and entitled “Anti-adenovirus antibodies, reagents and kits for detecting adenovirus,” the entire contents of which are incorporated by reference into the present disclosure. Technical Field

[0003] The present invention relates to the field of antibody technology, and in particular to an anti-adenovirus antibody, a reagent and a kit for detecting adenovirus. Background Art

[0004] Adenoviruses belong to the Adenoviridae family. They are non-enveloped viruses with a diameter of 65-80nM and a double-helix DNA genome. They are icosahedrons composed of 252 chitosans. Adenoviruses can exist stably in low pH environments, have strong resistance to physical and chemical agents, and can tolerate gastrointestinal secretions and bile. Therefore, adenoviruses can replicate in the gastrointestinal tract and produce a high viral load.

[0005] Adenovirus is the main pathogen causing pneumonia in children. Adenovirus pneumonia is one of the more serious types of community-acquired pneumonia in children. It is common in children aged 6 months to 5 years. Some children have severe clinical manifestations and many extrapulmonary complications. Severe cases are prone to chronic airway and lung diseases. It is currently one of the important causes of death and disability from pneumonia in infants and young children. In the past few years, adenovirus, as the main pathogen, often causes high morbidity and mortality in immunocompromised hosts such as AIDS patients, patients with genetic immunodeficiency, bone marrow recipients, solid organ and hematopoietic stem cell transplant recipients. Some fatal infections are often caused by adenovirus serotypes 1, 2, and 3, and microbial co-infections such as bacteria and fungi often occur in these patients. AIDS patients infected with adenovirus will develop complications such as pneumonia, hepatitis, meningeal softening, nephritis, and gastroenteritis. Therefore, it is extremely important for early diagnosis to detect adenovirus in a timely manner, achieve early detection and early treatment, and reduce the deterioration of the disease.

[0006] Adenovirus infection can be detected by serology and through respiratory and ocular secretions. Serological testing usually involves another antibody-antigen binding assay of specific antiserum. At present, the main detection methods of adenovirus are colloidal gold immunochromatography, which is an immunological detection method based on the specific reaction of antibodies and antigens, and uses colloidal gold to amplify and display the detected signal. Similar immunological detection methods include radioimmunoassay, enzyme-linked immunosorbent assay, chemiluminescence, etc. The above immunological detection methods all require antibodies against adenovirus. Therefore, there is a strong demand in the art for antibodies that are effective and bind to adenovirus and detect it.

[0007] In view of this, the present invention is proposed. Summary of the invention

[0008] The present application provides an anti-adenovirus antibody with enhanced activity to improve the detection of adenovirus and provide an important source of raw materials for the detection of adenovirus.

[0009] In order to achieve the above-mentioned object, according to one aspect of the present invention, an antibody or a functional fragment thereof is provided, wherein the antibody or the functional fragment thereof comprises three complementary determining regions having a heavy chain variable region of any one of the amino acid sequences SEQ ID NOs: 20 and 21 and three complementary determining regions having a light chain variable region of any one of the amino acid sequences SEQ ID NOs: 22, 23, 24, and 25.

[0010] In order to achieve the above-mentioned object, according to the second aspect of the present invention, an anti-adenovirus antibody or a functional fragment thereof is provided, wherein the antibody or the functional fragment thereof comprises HCDR1, HCDR2, HCDR3 having amino acid sequences as shown in SEQ ID NO:1 to SEQ ID NO:3, and LCDR1, LCDR2, LCDR3 having amino acid sequences as shown in SEQ ID NO:4 to SEQ ID NO:6.

[0011] In order to achieve the above-mentioned purpose, according to the third aspect of the present invention, an anti-adenovirus antibody or a functional fragment thereof is provided, wherein the antibody or the functional fragment thereof comprises a heavy chain variable region and / or a light chain variable region, the heavy chain variable region comprises a sequence structure of HFR1-HCDR1-HFR2-HCDR2-HFR3-HCDR3-HFR4, 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 amino acid sequences of the above-mentioned HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3.

[0012] In order to achieve the above-mentioned object, according to the fourth aspect of the present invention, an anti-adenovirus antibody or a functional fragment thereof is provided, wherein the antibody or the functional fragment thereof comprises a heavy chain variable region and / or a light chain variable region, and the amino acid sequence of the heavy chain variable region is shown in any one of SEQ ID NOs: 20 and 21; the amino acid sequence of the light chain variable region is shown in any one of SEQ ID NOs: 22, 23, 24, and 25.

[0013] In order to achieve the above object, according to the fifth aspect of the present invention, an anti-adenovirus antibody is provided, comprising a heavy chain and / or a light chain, wherein the heavy chain comprises the above-mentioned heavy chain variable region, and the light chain comprises the above-mentioned light chain variable region.

[0014] In order to achieve the above-mentioned object, according to the sixth aspect of the present invention, an anti-adenovirus antibody is provided, comprising a heavy chain and / or a light chain, wherein the amino acid sequence of the heavy chain is shown in any one of SEQ ID NOs: 26 and 27; the amino acid sequence of the light chain is shown in any one of SEQ ID NOs: 28, 29, 30, and 31.

[0015] In order to achieve the above object, according to the seventh aspect of the present invention, an antibody conjugate is provided, wherein the antibody conjugate comprises the above antibody or a functional fragment thereof.

[0016] In order to achieve the above object, according to the eighth aspect of the present invention, a reagent or kit for detecting adenovirus is provided, wherein the reagent or kit comprises the above antibody or its functional fragment or the above antibody conjugate.

[0017] In order to achieve the above object, according to the ninth aspect of the present invention, the present invention also provides a nucleic acid encoding the above antibody or a functional fragment thereof.

[0018] To achieve the above object, according to the tenth aspect of the present invention, a method for detecting adenovirus is provided, comprising: contacting the above antibody or its functional fragment with the adenovirus in a sample to be detected to form an immune complex.

[0019] In order to achieve the above object, according to the eleventh aspect of the present invention, a vector, a cell and a method for preparing the above antibody or a functional fragment thereof are provided.

[0020] In order to achieve the above-mentioned object, according to the twelfth aspect of the present invention, there is provided the use of the above-mentioned antibody or its functional fragment, antibody conjugate, reagent or kit in detecting or preparing adenovirus detection products. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the embodiments are briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without creative work.

[0022] Figure 1 The figure shows the results of reducing SDS-PAGE of Anti-ADV 3G9mut1 to mut6. DETAILED DESCRIPTION

[0023] In the first aspect, an embodiment of the present invention provides an antibody or a functional fragment thereof, wherein the antibody or the functional fragment thereof comprises three complementary determining regions of a heavy chain variable region having an amino acid sequence of any one of SEQ ID NOs: 20 and 21 and three complementary determining regions of a light chain variable region having an amino acid sequence of any one of SEQ ID NOs: 22, 23, 24, and 25.

[0024] It should be noted that HCDR1, HCDR2 and HCDR3 are amino acid sequences consistent with HCDR1, HCDR2 and HCDR3 of the same heavy chain variable region defined in the antibody or its functional fragment described in the first aspect, and LCDR1, LCDR2 and LCDR3 are amino acid sequences consistent with LCDR1, LCDR2 and LCDR3 of the same light chain variable region defined in the antibody or its functional fragment described in the first aspect.

[0025] For example, the HCDR1, HCDR2, and HCDR3 are amino acid sequences consistent with HCDR1, HCDR2, and HCDR3 of the heavy chain variable region shown in SEQ ID NO:20; the LCDR1, LCDR2, and LCDR3 are amino acid sequences consistent with LCDR1, LCDR2, and LCDR3 of the light chain variable region shown in SEQ ID NO:22.

[0026] 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.

[0027] In the present invention, the term "complementarity determining region", "CDR" or "CDRs" refers to the highly variable region of the heavy and light chains of immunoglobulins, and refers to the region containing one or more or even all of the major amino acid residues that play a role in the binding of an antibody or antigen-binding fragment to its recognized antigen or epitope. In a specific embodiment of the present invention, CDRs refers to the highly variable region of the heavy and light chains of the antibody.

[0028] In the present invention, the heavy chain complementarity determining region is represented by HCDR, which includes HCDR1, HCDR2 and HCDR3; the light chain complementarity determining region is represented by LCDR, which includes LCDR1, LCDR2 and LCDR3.

[0029] The definition methods of CDR are well known in the art, and the CDR definition methods include: Kabat definition, Chothia definition, IMGT definition, Contact definition and AbM definition. As described herein, "Kabat definition" refers to the definition system described in Kabat et al., US Pept. of Health and Human Services, "Sequence of Proteins of Immunological Interest" (1983). "Chothia definition" refers to Chothia et al., J Mol Biol 196: 901-917 (1987). There are other CDR definition methods that may not strictly follow one of the above schemes, but will still overlap with at least a portion of the CDR region defined by Kabat, although they may be shortened or extended based on predictions or experimental results of specific residues or residue groups. Exemplary defined CDRs are listed in Table 1 below, and the annotations in different documents are slightly different. Given the variable region amino acid sequence of an antibody, a person skilled in the art can routinely determine which residues contain a specific CDR. It should be noted that CDRs defined by other methods not limited to Table 1 also belong to the scope of protection of the present disclosure.

[0030] Table 1: CDR Definition 1

[0031] CDR Kabat <![CDATA[AbM 2 ]]> IMGT Chothia HCDR1 <![CDATA[H31~H35 3 ]]> <![CDATA[H26~H35 3 ]]> <![CDATA[H26~H33..5 5 ]]> <![CDATA[H26~H32..34 4 ]]> HCDR2 H50~H65 H50~H58 H51~H57 H52~H56 HCDR3 H95~H102 H95~H102 H93~H102 H95~H102 LCDR1 L24~L34 L24~L34 L27~L32 L24~L34 LCDR2 L50~L56 L50~L56 L50~L51 L50~L56 LCDR3 L89~L97 L89~L97 L89~L97 L89~L97

[0032] 1 The numbering of all CDR definitions in Table 1 is based on the Kabat numbering system (see below), with the amino acid numbering on the heavy chain represented by "H+number" and the amino acid numbering on the light chain represented by "L+number". One of ordinary skill in the art can unambiguously map the Kabat numbering system to any variable region sequence without relying on any experimental data other than the sequence itself. As used herein, "Kabat numbering" refers to the numbering system described by Kabat et al., U.S. Patent. of Health and Human Services, "Sequence of Proteins of Immunological Interest" (1983).

[0033] 2 "AbM" as used in Table 1 with a lowercase "b" refers to CDRs defined by Oxford Molecular's "AbM" antibody modeling software.

[0034] 3If both H35A and H35B are absent, CDR-H1 ends at position 35; if only H35A is present, CDR-H1 ends at position 35A; if both H35A and H35B are present, CDR-H1 ends at position 35B.

[0035] 4 If both H35A and H35B are absent, CDR-H1 ends at position 32; if only H35A is present, CDR-H1 ends at position 33; if both H35A and H35B are present, CDR-H1 ends at position 34.

[0036] 5 If both H35A and H35B are absent, CDR-H1 ends at position 33; if only H35A is present, CDR-H1 ends at position 34; if both H35A and H35B are present, CDR-H1 ends at position 35.

[0037] According to an embodiment of the present invention, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 or LCDR3 is defined by any one system or a combination of multiple systems of Kabat, Chothia, IMGT, AbM or Contact.

[0038] In some optional embodiments of the present invention, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 are defined by the Kabat system.

[0039] In some optional embodiments of the present invention, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 are defined by the Chothia system.

[0040] In some optional embodiments of the present invention, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 are defined by the IMGT system.

[0041] In some optional embodiments of the present invention, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 are defined by the AbM system.

[0042] In some optional embodiments of the present invention, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 are defined by a Contact system.

[0043] In some optional embodiments of the present invention, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 are defined by a combination of Kabat, Chothia, IMGT, AbM or Contact systems.

[0044] According to an embodiment of the present invention, the Kabat numbering positions corresponding to the amino acid sequences of HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 or LCDR3 defined by the Kabat, Chothia, AbM or IMGT systems are as follows:

[0045] CDR Kabat AbM IMGT Chothia HCDR1 H31~H35 H26~H35 H26~H33 H26~H32 HCDR2 H50~H65 H50~H58 H51~H57 H52~H56 HCDR3 H95~H102 H95~H102 H93~H102 H95~H102 LCDR1 L24~L34 L24~L34 L27~L32 L24~L34 LCDR2 L50~L56 L50~L56 L50~L51 L50~L56 LCDR3 L89~L97 L89~L97 L89~L97 L89~L97

[0046] According to an embodiment of the present invention, the HCDR1, HCDR2 and HCDR3 include or are the amino acid sequences at positions 31-35A, 50-65, 95-99 of the heavy chain variable region numbered by Kabat, respectively; the LCDR1, LCDR2 and LCDR3 include or are the amino acid sequences at positions 24-34, 50-56, 89-95 of the light chain variable region numbered by Kabat, respectively.

[0047] In a second aspect, the present invention provides an anti-adenovirus antibody or a functional fragment thereof, wherein the antibody or the functional fragment thereof comprises HCDR1, HCDR2, HCDR3 with amino acid sequences as shown in SEQ ID NO: 1 to SEQ ID NO: 3, and LCDR1, LCDR2, LCDR3 with amino acid sequences as shown in SEQ ID NO: 4 to SEQ ID NO: 6. The above-mentioned antibody has improved activity.

[0048] In the present invention, the "framework region" or "FR" region includes the heavy chain framework region and the light chain framework region, and refers to the region other than the CDR in the heavy chain variable region and the light chain variable region of the antibody; wherein the heavy chain framework region can be further subdivided into adjacent regions separated by CDR, including HFR1, HFR2, HFR3 and HFR4 framework regions; the light chain framework region can be further subdivided into adjacent regions separated by CDR, including LFR1, LFR2, LFR3 and LFR4 framework regions.

[0049] 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.

[0050] In an optional embodiment, the antibody or a functional fragment thereof further has at least one of HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3 and LFR4;

[0051] The HFR1 comprises SEQ ID NO:7 or an amino acid sequence having at least 80% homology thereto;

[0052] The HFR2 comprises SEQ ID NO: 8 or an amino acid sequence having at least 80% homology thereto;

[0053] The HFR3 comprises SEQ ID NO: 9 or an amino acid sequence having at least 80% homology thereto;

[0054] The HFR4 comprises SEQ ID NO: 10 or an amino acid sequence having at least 80% homology thereto;

[0055] The LFR1 comprises SEQ ID NO: 11 or an amino acid sequence having at least 80% homology thereto;

[0056] The LFR2 comprises SEQ ID NO: 12 or an amino acid sequence having at least 80% homology thereto;

[0057] The LFR3 comprises SEQ ID NO: 13 or an amino acid sequence having at least 80% homology thereto;

[0058] The LFR4 comprises SEQ ID NO: 14 or an amino acid sequence having at least 80% homology thereto;

[0059] In an optional embodiment, the antibody or its functional fragment further comprises HFR1, HFR2, HFR3, HFR4 having amino acid sequences as shown in SEQ ID NO:7 to SEQ ID NO:10 and LFR1, LFR2, LFR3 and LFR4 having amino acid sequences as shown in SEQ ID NO:11 to SEQ ID NO:14.

[0060] It should be noted that, in other embodiments, the amino acid sequences of the framework regions of the antibodies or functional fragments thereof provided by the present invention 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 corresponding framework regions (SEQ ID NO: 7, 8, 9, 10, 11, 12, 13 or 14).

[0061] In an optional embodiment, the amino acid sequence of HFR1 of the antibody or its functional fragment is as shown in SEQ ID NO:15.

[0062] In an optional embodiment, the amino acid sequence of LFR3 of the antibody or its functional fragment is as shown in SEQ ID NO:16.

[0063] In an optional embodiment, the amino acid sequence of LFR4 of the antibody or its functional fragment is as shown in SEQ ID NO:17.

[0064] In a third aspect, an embodiment of the present invention provides an anti-adenovirus antibody or a functional fragment thereof, wherein the antibody or the functional fragment thereof comprises a heavy chain variable region and / or 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, and the amino acid sequences of the HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3, and LFR4 are the above-mentioned amino acid sequences of HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3, and LFR4.

[0065] In an optional embodiment, the heavy chain variable region amino acid sequence is shown in any one of SEQ ID NOs: 20 to 21.

[0066] In an optional embodiment, the light chain variable region amino acid sequence is shown in any one of SEQ ID NOs: 22 to 25.

[0067] In an optional embodiment, the antibody or functional fragment thereof described in the first aspect, the second aspect, or the third aspect comprises any combination of the following heavy chain variable regions and light chain variable regions:

[0068] combination Heavy chain variable region Light chain variable region 1 SEQ ID NO:20 SEQ ID NO:22 2 SEQ ID NO:21 SEQ ID NO:22 3 SEQ ID NO:20 SEQ ID NO:24 4 SEQ ID NO:20 SEQ ID NO:23 5 SEQ ID NO:21 SEQ ID NO:23 6 SEQ ID NO:20 SEQ ID NO:25

[0069] In an alternative embodiment, the antibody further comprises a constant region.

[0070] In an alternative embodiment, the constant region includes a heavy chain constant region and / or a light chain constant region.

[0071] In an alternative embodiment, the heavy chain constant region is selected from the heavy chain constant region of IgG1, IgG2, IgG3, IgG4, IgA, IgM, IgE or IgD, and the light chain constant region is selected from the κ type or λ type light chain constant region.

[0072] In an alternative embodiment, the constant region is derived from a species of cow, horse, dairy cow, pig, sheep, rat, mouse, dog, cat, rabbit, donkey, deer, mink, chicken, duck, goose, turkey, fighting cock or human.

[0073] In an alternative embodiment, the species origin of the constant region is mouse.

[0074] In an optional embodiment, the heavy chain constant region sequence (CH) is as shown in SEQ ID NO:18, and the light chain constant region (CL) sequence is as shown in SEQ ID NO:19.

[0075] It should be noted that, in other embodiments, the constant region sequence may have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with the above-mentioned constant region (SEQ ID NO: 18 or 19).

[0076] In an optional embodiment, the functional fragment is selected from any one of F(ab')2, Fab', Fab, Fv and scFv of the antibody.

[0077] The functional fragments of the above antibodies usually have the same binding specificity as the antibodies from which they are derived. It is easy for a person skilled in the art to understand based on the contents described in the present invention that the functional fragments of the above antibodies can be obtained by methods such as enzymatic digestion (including pepsin or papain) and / or by chemical reduction to split disulfide bonds. Based on the structure of the complete antibody disclosed in the present invention, a person skilled in the art can easily obtain the above functional fragments.

[0078] The functional 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 and the like.

[0079] In a fourth aspect, the present invention provides an anti-adenovirus antibody, comprising a heavy chain and / or a light chain, wherein the heavy chain comprises the above-mentioned heavy chain variable region and the above-mentioned heavy chain constant region; and the light chain comprises the above-mentioned light chain variable region and the above-mentioned light chain constant region.

[0080] In an optional embodiment, the amino acid sequence of the heavy chain is as shown in any one of SEQ ID NOs: 26 to 27.

[0081] In an optional embodiment, the amino acid sequence of the light chain is as shown in any one of SEQ ID NOs: 28 to 31.

[0082] In an optional embodiment, the antibody described in the first aspect, the second aspect, the third aspect, and the fourth aspect comprises any combination of the following heavy chains and light chains:

[0083] combination Heavy chain Light chain 1 SEQ ID NO:26 SEQ ID NO:28 2 SEQ ID NO:27 SEQ ID NO:28 3 SEQ ID NO:26 SEQ ID NO:30 4 SEQ ID NO:26 SEQ ID NO:29 5 SEQ ID NO:27 SEQ ID NO:29 6 SEQ ID NO:26 SEQ ID NO:31

[0084] In a fifth aspect, the present invention provides an antibody conjugate, comprising the above-mentioned antibody, wherein the antibody is directly or indirectly covalently coupled to the object to be coupled, or the antibody is coupled to the object to be coupled by non-covalent adsorption.

[0085] In an alternative embodiment, the antibody in the above antibody conjugate is conjugated to biotin or a biotin derivative.

[0086] In an alternative embodiment, the antibody in the above antibody conjugate is conjugated to a label.

[0087] 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.

[0088] In an alternative embodiment, the label includes but is not limited to fluorescent dyes, enzymes, radioactive isotopes, chemiluminescent agents and nanoparticle labels.

[0089] 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 belongs to the protection scope of the present invention.

[0090] In an optional embodiment, the fluorescent dye is 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, Cy3.7, Cy3.8, Cy3.9, Cy3.10, Cy3.11, Cy3.12, Cy3.13, Cy3.14, Cy3.15, Cy3.16, Cy3.17, Cy3.18, Cy3.19, Cy3.20, Cy3.21, Cy3.22, Cy3.23, Cy3.24, Cy3.25, Cy3.26, Cy3.27, Cy3.28, Cy3.29, Cy3.30, Cy3.31, Cy3.32, Cy3.33, Cy3.34, Cy3.35, Cy3.36, Cy3.37, Cy3.38, Cy3.39, Cy3.40, Cy3.41, Cy3.42, Cy3.43, Cy3.44, Cy3.45, Cy3.46, Cy3.47, Cy3.48, Cy3.49, Cy3.50, Cy3.51, Cy3.52, Cy3.53, Cy3.54, Cy3.55, Cy3.56, Cy3.57, Cy3.58, Cy3.59, Cy3.59, Cy3.59, Cy3.59, Cy3.59, Cy3.5 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), pre-chlorophyll protein (preCP), etc.).

[0091] In alternative embodiments, the enzyme includes, but is not limited to, horseradish peroxidase, alkaline phosphatase, β-galactosidase, glucose oxidase, carbonic anhydrase, acetylcholinesterase, and 6-phosphoglucose deoxygenase.

[0092] 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.

[0093] In an optional embodiment, the chemiluminescent reagent includes but is not limited to luminol and its derivatives, lucigenin, crustacean fluorescein and its derivatives, bipyridine ruthenium and its derivatives, acridinium esters and their derivatives, dioxetanes and their derivatives, lophanes and their derivatives, and peroxyoxalates and their derivatives.

[0094] 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.

[0095] In alternative embodiments, the colloid includes, but is not limited to, colloidal metals, disperse dyes, dye-labeled microspheres, and latex.

[0096] In an alternative embodiment, the colloidal metal includes, but is not limited to, colloidal gold, colloidal silver, and colloidal selenium.

[0097] In an optional embodiment, the colloidal metal is colloidal gold.

[0098] In an alternative embodiment, the antibody in the above antibody conjugate is coupled to a solid phase.

[0099] In alternative embodiments, the solid phase is selected from the group consisting of microspheres, plates, and membranes.

[0100] 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.

[0101] In an alternative embodiment, the solid phase is a nitrocellulose membrane.

[0102] In a sixth aspect, the present invention provides a reagent or a kit for detecting adenovirus, wherein the reagent or the kit comprises the above-mentioned antibody or a functional fragment thereof or the above-mentioned antibody conjugate.

[0103] In a seventh aspect, the present invention provides use of the above-mentioned antibody or its functional fragment, antibody conjugate or the above-mentioned reagent or kit in adenovirus detection.

[0104] In an eighth aspect, the present invention provides a method for detecting adenovirus, comprising: contacting the above-mentioned antibody or a functional fragment thereof with the adenovirus in a sample to be detected to form an immune complex.

[0105] In an optional embodiment, the immune complex further comprises a second antibody, which binds to the antibody or a functional fragment thereof.

[0106] In an alternative embodiment, the immune complex further comprises a second antibody, which binds to the adenovirus.

[0107] In a ninth aspect, the present invention provides a nucleic acid molecule encoding the above-mentioned antibody or a functional fragment thereof.

[0108] In a tenth aspect, the present invention provides a vector containing the above-mentioned nucleic acid molecule.

[0109] In an eleventh aspect, the present invention provides a cell containing the above-mentioned vector.

[0110] In a twelfth aspect, the present invention provides a method for preparing an antibody or a functional fragment thereof, comprising: culturing the cells as described above.

[0111] In a thirteenth aspect, the present invention provides use of the above-mentioned antibodies or functional fragments thereof, antibody conjugates, reagents or kits in detecting or preparing adenovirus detection products.

[0112] Based on the amino acid sequence of the antibody or its functional fragment disclosed in the present invention, those skilled in the art can easily think of using genetic engineering technology or other technologies (chemical synthesis, recombinant expression) to prepare the antibody or its functional fragment, for example, separating and purifying the antibody or its functional fragment from the culture product of a recombinant cell that can recombinantly express the antibody or its functional fragment as described in any of the above items, which is easy to achieve for those skilled in the art. Based on this, no matter what technology is used to prepare the antibody or its functional fragment of the present invention, it belongs to the protection scope of the present invention.

[0113] In order to make the purpose, technical scheme and advantages of the embodiments of the present invention clearer, the technical scheme in the embodiments of the present invention will be described clearly and completely below. If the specific conditions are not specified in the embodiments, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

[0114] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those of ordinary skill in the art to which the present disclosure belongs. Although any methods and materials similar or equivalent to those described herein can be used for the practice or testing of the preparations or unit doses herein, some methods and materials are now described. Unless otherwise stated, the techniques adopted or considered herein are standard methods. Materials, methods and examples are illustrative and non-restrictive only.

[0115] 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 skilled artisan. This technique is fully explained in the literature, such as Molecular Cloning: A Laboratory Manual, 2nd Edition (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); PCR: The Polymerase Chain Reaction (PCR: The Polymerase Chain Reaction) (Academic Press, Inc., 1987). 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.

[0116] The features and performance of the present invention are further described in detail below in conjunction with the embodiments.

[0117] Example 1 Preparation of Anti-ADV 3G9 Monoclonal Antibody

[0118] In this example, restriction endonucleases and Prime Star DNA polymerase were purchased from Takara. MagExtractor-RNA extraction kit was purchased from TOYOBO. BD SMART TMRACE cDNA Amplification Kit was purchased from Takara. pMD-18T vector was purchased from Takara. Plasmid extraction kit was purchased from Tiangen. Primer synthesis and gene sequencing were completed by Invitrogen.

[0119] 1 Construction of recombinant plasmid

[0120] (1) Antibody gene preparation

[0121] mRNA was extracted from the hybridoma cell line that secreted anti-adenovirus monoclonal antibodies, and the DNA product was obtained by RT-PCR. The product was inserted into the pMD-18T vector after A addition reaction with rTaq DNA polymerase and transformed into DH5α competent cells. After the colonies grew, the Heavy Chain and Light Chain gene clones were taken respectively, and 4 clones each were sent to a gene sequencing company for sequencing.

[0122] (2) Sequence analysis of the variable region gene of Anti-ADV 3G9 antibody

[0123] The gene sequences obtained by the above sequencing were placed in the Kabat antibody database for analysis, and the 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 the gene fragments amplified by the Light Chain, the VL gene sequence was 321 bp, and was preceded by a 57 bp leader peptide sequence; among the gene fragments amplified by the Heavy Chain primer pair, the VH gene sequence was 351 bp, belonging to the VH1 gene family, and was preceded by a 57 bp leader peptide sequence.

[0124] (3) Construction of recombinant antibody expression plasmid

[0125] pcDNA TM 3.4 vector is a recombinant antibody eukaryotic expression vector constructed, which has introduced multiple cloning restriction sites such as HindIII, BamHI, and EcoRI, and is named pcDNA3.4A expression vector, hereinafter referred to as 3.4A expression vector; according to the sequencing results of the antibody variable region genes in the above pMD-18T, the VL and VH gene-specific primers of the antibody were designed, with HindIII, EcoRI restriction sites and protective bases at both ends, respectively, and the 0.72kb Light Chain gene fragment and the 1.40kb Heavy Chain gene fragment were amplified by the PCR amplification method.

[0126] The Heavy Chain and Light Chain gene fragments were double-digested with HindIII / EcoRI, and the 3.4A vector was double-digested with HindIII / EcoRI. After the fragments and vectors were purified and recovered, the Heavy Chain gene and the 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.

[0127] 2. Screening of stable cell lines

[0128] (1) Transient transfection of recombinant antibody expression plasmid into CHO cells to determine the activity of the expression plasmid

[0129] The plasmid was diluted to 40ug / 100ul with ultrapure water and the CHO cells were adjusted to 1.43×10 7 cells / ml in a centrifuge tube, mix 100 μL of plasmid with 700 μL of cells, transfer to an electroporation cup, electroporate, take samples and count on the 3rd, 5th and 7th days, and collect samples for detection on the 7th day.

[0130] The coating solution (main component NaHCO3) was used to dilute adenovirus (purchased from microbix, item number EL-15-02) to 300 times, 100 μL per well, and incubated at 4°C overnight; the next day, the cells were washed twice with the washing solution and patted dry; the blocking solution (20% BSA + 80% PBS) was added, 120 μL per well, incubated at 37°C for 1 hour, and patted dry; the diluted cell supernatant was added, 100 μL / well, incubated at 37°C for 30 minutes (part of the supernatant was removed for 1 hour); the washing solution was used to wash the cells twice and the cells were patted dry. Wash 5 times and pat dry; add goat anti-mouse IgG-HRP, 100 μL per well, 37°C, 30 min; wash 5 times with washing solution and pat dry; add color development solution A (50 μL / well, containing citric acid + sodium acetate + acetanilide + urea peroxide), add color development solution B (50 μL / well, containing citric acid + EDTA·2Na + TMB + concentrated HCL), 10 min; add stop solution (50 μL / well, containing EDTA·2Na + concentrated HCL) 2 SO 4 ); read the OD value at 450nm (reference 630nm) on the microplate reader. The results showed that the reaction OD was still greater than 1.0 after the cell supernatant was diluted 1000 times, and the reaction OD of the well without cell supernatant was less than 0.1, indicating that the antibodies produced after transient plasmid transfection were active against adenovirus.

[0131] (2) Linearization of recombinant antibody expression plasmid

[0132] Prepare the following reagents: 50 μL of Buffer, 100 μg of DNA / tube, 10 μL of PuvⅠ enzyme, and make up to 500 μL with sterile water. Perform enzymatic digestion in a 37°C water bath overnight. First, use an equal volume of phenol / chloroform / isoamyl alcohol (lower layer) 25:24:1, and then use chloroform (aqueous phase) for extraction in sequence. Precipitate on ice with 0.1 volume (aqueous phase) of 3M sodium acetate and 2 volumes of ethanol, rinse the precipitate with 70% ethanol, remove the organic solvent, and reconstitute with an appropriate amount of sterile water after the ethanol has completely evaporated. Finally, measure the concentration.

[0133] (3) Stable transfection of recombinant antibody expression plasmid and pressure screening of stable cell lines

[0134] The plasmid was diluted to 40ug / 100ul with ultrapure water and the CHO cells were adjusted to 1.43×10 7 cells / ml in a centrifuge tube, mix 100μL plasmid with 700μL cells, transfer to an electroporation cup, electrotransform, and count the next day; culture under pressure in 25umol / L MSX 96-well for about 25 days.

[0135] Observe the clone wells with cells under a microscope and record the confluence; take the culture supernatant and send it for testing; select cell lines with high antibody concentration and relative concentration and transfer them to 24 wells, and transfer them to 6 wells after about 3 days; after 3 days, keep the seed batch culture and adjust the cell density to 0.5×10 6 cells / ml, 2.2 ml for batch culture, cell density 0.3×10 6 cells / ml, 2ml for seed preservation; 7-day 6-well batch culture supernatant samples were sent for testing, and cell lines with smaller antibody concentrations and cell diameters were selected for TPP seed preservation and subculture.

[0136] 3. Recombinant Antibody Production

[0137] (1) Cell expansion

[0138] After cell recovery, the cells are first cultured in a 125ml shake flask with an inoculation volume of 30ml and a culture medium of 100% Dynamis medium. The cells are placed in a shaker at a speed of 120r / min, a temperature of 37°C, and a carbon dioxide content of 8%. After 72 hours of culture, the cells are inoculated and expanded at an inoculation density of 500,000 cells / ml. The expansion volume is calculated according to production requirements, and the culture medium is 100% Dynamis medium. The cells are expanded every 72 hours. When the cell volume meets the production requirements, the inoculation density is strictly controlled to be around 500,000 cells / ml for production.

[0139] (2) Shake flask production and purification

[0140] Shake flask parameters: speed 120r / min, temperature 37℃, carbon dioxide 8%. Feeding: Start feeding every day when the culture reaches 72h in the shake flask. HyCloneTM Cell BoostTM Feed 7a is fed daily at 3% of the initial culture volume, and Feed 7b is fed daily at one thousandth of the initial culture volume until the 12th day (feeding on the 12th day). Glucose is added at 3g / L on the sixth day. Samples are collected on the 13th day. Affinity purification is performed using a proteinA affinity chromatography column. Take 3μg of the purified antibody for reducing SDS-PAGE. The electrophoresis is shown in the figure. After reducing SDS-PAGE, two bands are shown, one Mr is 50KD and the other Mr is 28KD.

[0141] Example 2 Activity Optimization

[0142] Although the Anti-ADV 3G9 monoclonal antibody obtained in Example 1 has the ability to bind to adenovirus, the antibody activity is not ideal, so the applicant carried out directed mutation of the light chain CDR and heavy chain CDR of the antibody. That is, the computer is used to simulate the structure of the antibody variable region, the structure of the complex of the antigen and the antibody variable region, the key amino acid analysis of the antibody and the mutation design, and the bidirectional primers covering the mutation site are designed and synthesized according to the mutation scheme, the primers at both ends of the target DNA are synthesized, and a high-fidelity PCR reaction is performed. The PCR product is cloned into a vector, and then the mutant antibody is prepared according to the method described in Example 1. After screening, monoclonal antibodies with significantly improved antibody activity were obtained and named: Anti-ADV 3G9mut1 to Anti-ADV 3G9mut6. The heavy chain and light chain amino acid sequences of each monoclonal antibody are shown in the following table.

[0143] Table 2 Antibody sequences

[0144] Sample name Heavy chain number Light chain number Anti-ADV 3G9mut1 SEQ ID NO:26 SEQ ID NO:28 Anti-ADV 3G9mut2 SEQ ID NO:27 SEQ ID NO:28 Anti-ADV 3G9mut3 SEQ ID NO:26 SEQ ID NO:30 Anti-ADV 3G9mut4 SEQ ID NO:26 SEQ ID NO:29 Anti-ADV 3G9mut5 SEQ ID NO:27 SEQ ID NO:29 Anti-ADV 3G9mut6 SEQ ID NO:26 SEQ ID NO:31

[0145] Example 3 Performance Testing of Antibodies

[0146] 1. Activity identification

[0147] The coating solution (main component NaHCO3) was used to dilute adenovirus (purchased from microbix, item number EL-15-02) to 300 times, 100 μL per well, and incubated at 4°C overnight; the next day, the cells were washed twice with the washing solution and patted dry; the blocking solution (20% BSA + 80% PBS) was added, 120 μL per well, incubated at 37°C for 1 hour, and patted dry; the diluted monoclonal antibodies were added, 100 μL / well, incubated at 37°C for 30-60 minutes; the cells were washed 5 times with the washing solution and patted dry. Dry; add goat anti-mouse IgG-HRP, 100 μl per well, 37°C, 30 min; wash with washing solution (PBS) 5 times, pat dry; add color development solution A (50 μl / well, containing 2.1g / L citric acid, 12.25g / L citric acid, 0.07g / L acetanilide and 0.5g / L urea peroxide), add color development solution B (50 μl / well, containing 1.05g / L citric acid, 0.186g / EDTA·2Na, 0.45g / L TMB and 0.2ml / L concentrated HCl), 10 min; add stop solution (50 μl / well, containing 0.75g / EDTA·2Na and 10.2ml / L concentrated HCl 2 SO 4 ); read the OD value at 450nm (reference 630nm) on the microplate reader. The results are shown in the table below.

[0148] Table 3 Activity data

[0149] Sample concentration (ng / ml) 125 62.5 31.25 15.625 7.813 0 Comparison 1.443 1.068 0.698 0.411 0.208 0.032 Anti-ADV 3G9mut1 1.739 1.418 1.029 0.674 0.389 0.029 Anti-ADV 3G9mut2 1.786 1.44 1.071 0.685 0.406 0.028 Anti-ADV 3G9mut3 1.721 1.416 1.068 0.677 0.395 0.022 Anti-ADV 3G9mut4 1.819 1.48 1.074 0.693 0.403 0.023 Anti-ADV 3G9mut5 1.655 1.382 1.025 0.643 0.362 0.031 Anti-ADV 3G9mut6 1.718 1.397 1.044 0.68 0.386 0.022

[0150] 2. Stability assessment

[0151] The above antibodies were placed at 4°C (refrigerator), -80°C (refrigerator), and 37°C (constant temperature box) for 21 days, and samples were taken for 7 days, 14 days, and 21 days for status observation, and the activity of the 21-day samples was tested. The results showed that there was no obvious protein state change in the antibodies under the three test conditions for 21 days, and the activity did not show a downward trend with the increase of the test temperature, indicating that the above antibodies are stable. Table 4 below shows the OD results of the enzyme immunoassay activity test for 21 days of antibody test.

[0152] Table 4 Stability data

[0153] Sample concentration (ng / ml) 125 15.625 0 4℃, 21-day samples 1.722 0.651 0.033 -80℃, 21 days sample 1.740 0.669 0.022 37℃, 21 days sample 1.701 0.632 0.035

[0154] 3. Performance Evaluation

[0155] The above antibody was used as coating and another strain of ADV antibody was used as labeling. The performance was verified on the colloidal gold platform as shown in the following table:

[0156] Note: B: negative, C: positive, C1>C1+>C2>C2+>C3>C3+, and so on. The larger the number, the lighter the color.

[0157] Table 5 Performance evaluation data

[0158]

[0159] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

[0160] The partial amino acid sequences involved in this application are as follows:

[0161]

[0162]

[0163]

Claims

1. An anti-adenovirus antibody or an antigen-binding fragment thereof, wherein the antibody or the 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 of the heavy chain variable region shown in any one of SEQ ID NOs: 20 and 21, and the LCDR1, LCDR2, and LCDR3 combination is the same as the LCDR1, LCDR2, and LCDR3 combination of the light chain variable region shown in any one of SEQ ID NOs: 22, 23, 24, and 25; the HCDR1, HCDR2, HCDR3 and LCDR1, LCDR2, and LCDR3 are numbered or defined by the Kabat, Chothia, IMGT, Lesk, AbM, or Contact systems.

2. An anti-adenovirus antibody or an antigen-binding fragment thereof, characterized in that: The antibody or its antigen-binding fragment comprises HCDR1, HCDR2, HCDR3 whose amino acid sequences are shown in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, and LCDR1, LCDR2, LCDR3 whose amino acid sequences are shown in SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, respectively.

3. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 2, characterized in that: The antibody or antigen-binding fragment thereof further has at least one of HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3 and LFR4; The HFR1 comprises SEQ ID NO:7 or an amino acid sequence having at least 80% homology thereto; The HFR2 comprises SEQ ID NO: 8 or an amino acid sequence having at least 80% homology thereto; The HFR3 comprises SEQ ID NO: 9 or an amino acid sequence having at least 80% homology thereto; The HFR4 comprises SEQ ID NO: 10 or an amino acid sequence having at least 80% homology thereto; The LFR1 comprises SEQ ID NO: 11 or an amino acid sequence having at least 80% homology thereto; The LFR2 comprises SEQ ID NO: 12 or an amino acid sequence having at least 80% homology thereto; The LFR3 comprises SEQ ID NO: 13 or an amino acid sequence having at least 80% homology thereto; The LFR4 comprises SEQ ID NO: 14 or an amino acid sequence having at least 80% homology thereto.

4. The antibody or antigen-binding fragment thereof according to any one of claims 1-2, wherein the antibody or antigen-binding fragment thereof further comprises HFR1, HFR2, HFR3, HFR4 whose amino acid sequences are shown in SEQ ID NOs: 7, 8, 9, 10, and LFR1, LFR2, LFR3, and LFR4 whose amino acid sequences are shown in SEQ ID NOs: 11, 12, 13, 14, or an amino acid sequence that is at least 80% identical to said sequences.

5. The antibody or antigen-binding fragment thereof according to claim 3, characterized in that: The amino acid sequence of HFR1 of the antibody or antigen-binding fragment thereof is shown in SEQ ID NO:

15.

6. The antibody or antigen-binding fragment thereof according to claim 3, characterized in that: The amino acid sequence of LFR3 of the antibody or antigen-binding fragment thereof is shown in SEQ ID NO:

16.

7. The antibody or antigen-binding fragment thereof according to claim 3, characterized in that: The amino acid sequence of LFR4 of the antibody or antigen-binding fragment thereof is shown in SEQ ID NO:

17.

8. An anti-adenovirus antibody or antigen-binding fragment thereof, characterized in that: The antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein the amino acid sequence of the heavy chain variable region is shown in any one of SEQ ID NOs: 20 and 21; the amino acid sequence of the light chain variable region is shown in any one of SEQ ID NOs: 22, 23, 24, and 25.

9. The antibody or antigen-binding fragment thereof according to any one of claims 1-2, 5-8, characterized in that: The antibody or antigen-binding fragment thereof further comprises a constant region.

10. The antibody or antigen-binding fragment thereof according to claim 9, characterized in that: The constant region includes a heavy chain constant region and / or a light chain constant region.

11. The antibody or antigen-binding fragment thereof according to claim 10, characterized in that: The heavy chain constant region is selected from the heavy chain constant region of IgG1, IgG2, IgG3, IgG4, IgA, IgM, IgE or IgD; the light chain constant region is selected from the κ type or λ type light chain constant region.

12. The antibody or antigen-binding fragment thereof according to claim 9, characterized in that: The species origin of the constant region is cow, horse, pig, sheep, goat, rat, mouse, dog, cat, rabbit, donkey, deer, mink, chicken, duck, goose or human.

13. The antibody or antigen-binding fragment thereof according to claim 9, characterized in that: The species origin of the constant region is mouse.

14. The antibody or antigen-binding fragment thereof according to claim 10, characterized in that: The heavy chain constant region sequence is as shown in SEQ ID NO: 18 or has at least 80% identity thereto.

15. The antibody or antigen-binding fragment thereof according to claim 10, characterized in that: The light chain constant region sequence is as shown in SEQ ID NO: 19 or has at least 80% identity thereto.

16. The antibody or antigen-binding fragment thereof according to any one of claims 1-2, 5-8, 10-15, characterized in that: The antigen-binding fragment is selected from any one of F(ab')2, Fab', Fab, Fv and scFv of the antibody.

17. An anti-adenovirus antibody or antigen-binding fragment thereof, comprising a heavy chain and a light chain, characterized in that: The amino acid sequence of the heavy chain is shown in any one of SEQ ID NOs: 26 and 27; the amino acid sequence of the light chain is shown in any one of SEQ ID NOs: 28, 29, 30, and 31.

18. An antibody conjugate, characterized in that: The antibody conjugate comprises the antibody or antigen-binding fragment thereof according to any one of claims 1 to 17.

19. The antibody conjugate according to claim 18, characterized in that The antibody is conjugated to biotin or a biotin derivative.

20. The antibody conjugate according to claim 18, characterized in that The antibody is coupled to a solid phase.

21. The antibody conjugate according to claim 18, characterized in that The antibody is conjugated to a label.

22. The antibody conjugate according to claim 21, characterized in that The label is selected from fluorescent dyes, enzymes, radioisotopes, chemiluminescent reagents and nanoparticle labels.

23. The antibody conjugate according to claim 21, characterized in that The label is colloidal gold.

24. A reagent or kit for detecting adenovirus, characterized in that: The reagent or kit comprises the antibody or antigen-binding fragment thereof according to any one of claims 1 to 17 or the antibody conjugate according to any one of claims 18 to 23.

25. Use of the antibody or antigen-binding fragment thereof according to any one of claims 1 to 17, the antibody conjugate according to any one of claims 18 to 23, or the reagent or kit according to claim 24 in the preparation of a product for detecting adenovirus, characterized in that: include: The antibody or antigen-binding fragment thereof according to any one of claims 1 to 17 is contacted with the adenovirus in the sample to be detected to form an immune complex.

26. The use according to claim 25, characterized in that The immune complex also includes a second antibody that binds to the antibody or antigen-binding fragment thereof.

27. The use according to claim 25, characterized in that The immune complex also includes a second antibody that binds to the adenovirus.

28. A nucleic acid, characterized in that It encodes the antibody or antigen-binding fragment thereof according to any one of claims 1 to 17.

29. A carrier, characterized in that It contains the nucleic acid according to claim 28.

30. A cell, characterized in that It contains the nucleic acid according to claim 28 or the vector according to claim 29.

31. A method for preparing the antibody or antigen-binding fragment thereof according to any one of claims 1 to 17, characterized in that: It includes: Cultivate the cell of claim 30.

32. Use of the antibody or antigen-binding fragment thereof according to any one of claims 1 to 17, or the antibody conjugate according to any one of claims 18 to 23, or the reagent or kit according to claim 24 in the preparation of a product for detecting adenovirus.

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