Anti-human type 4 adenovirus specific humanized antibody E2DG and application thereof in antiviral therapy

Abstract

The invention provides a human monoclonal antibody E2DG for resisting type 4 adenovirus. The neutralizing antibody for resisting the 4-type adenovirus provided by the invention can be specifically combined with the 4-type adenovirus, and has very high affinity activity and very strong neutralizing activity at low concentration. The invention also provides application of the antibody in preparation of medicines for treating and / or preventing type 4 adenovirus diseases, and a detection kit containing the antibody.

Description

TECHNICAL FIELD

[0001] The application discloses a monoclonal antibody and belongs to the technical field of immunology and microbiology. BACKGROUND

[0002] Human adenovirus (HAdV) is a DNA virus that often causes respiratory tract infection, and is divided into seven subgroups A-G. Among them, B, C and E can cause upper respiratory tract infection, lower respiratory tract infection and bronchitis and other respiratory diseases.

[0003] Adenovirus infection is widely prevalent worldwide, is prone to occur in spring and winter, spreads rapidly and infects people of a wide age range, and is particularly prone to occur in children. Adenovirus patients and asymptomatic infected persons are the main sources of infection of adenovirus, and outbreak infection often occurs in closed or crowded places. In a large-scale epidemic, most patients show mild self-limiting infection, and some cases are also accompanied by gastrointestinal symptoms such as diarrhea, vomiting and abdominal pain, and severe cases can lead to pneumonia and even death. Among them, pneumonia caused by adenovirus infection is particularly serious, and the mortality rate of patients with severe HAdV pneumonia or diffuse disease without treatment can be more than 50%. Severe pneumonia patients not only show symptoms such as heart failure, respiratory failure, encephalitis, liver damage, myocarditis or myocardial damage, and even death, and the effect of intervention through related means is still poor.

[0004] The pathogenic factors of adenovirus are composed of a variety of major and minor capsid proteins, and Hexon, Penton and Fiber are three major capsid proteins. Among them, Hexon protein is the most abundant in the adenovirus capsid and constitutes the main surface structure of the virus particle. The three major capsid proteins on the surface of the virus capsid all have certain immunogenicity and can stimulate the body to produce neutralizing antibodies, and the Hexon protein is the main protein for forming the adenovirus capsid and producing specific antibodies, which can induce the host to produce a strong immune response, including producing neutralizing antibodies and initiating a cellular immune response.

[0005] In summary, there is no effective vaccine and drug for preventing and treating HAdV pneumonia, especially pneumonia caused by type 4 adenovirus of group E.

[0006] With the development of genetic engineering, single-cell sequencing technology has become a revolutionary tool for antibody discovery. High-efficiency, natural, fully human monoclonal antibodies are increasingly showing advantages. Their safety, effectiveness and relevance to human diseases have been significantly improved compared with polyclonal antibodies or murine monoclonal antibodies obtained by traditional antibody discovery techniques. The purpose of the present application is to provide a natural, fully human monoclonal neutralizing antibody against type 4 adenovirus using single-cell sequencing technology, and to provide its application in the preparation of adenovirus prevention or treatment drugs. SUMMARY

[0007] Based on the above purpose, the present application first provides a natural human monoclonal antibody against type 4 adenovirus Hexon protein. The amino acid sequences of CDR1, CDR2 and CDR3 regions of the heavy chain variable region of the natural human monoclonal antibody against type 4 adenovirus are respectively shown in SEQ ID NO: 1 26-33, 51-58 and 97-116; the amino acid sequences of CDR1, CDR2 and CDR3 regions of the light chain variable region are respectively shown in SEQ ID NO: 3 26-36, 54-56 and 93-101.

[0008] In a preferred embodiment, the amino acid sequence of the heavy chain variable region of the natural human monoclonal antibody against type 4 adenovirus is shown in SEQ ID NO: 1, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 3. In the present application, one specific embodiment with the heavy chain variable region and the light chain variable region is named as "E2DG" antibody.

[0009] In a preferred embodiment, the amino acid sequence of the heavy chain constant region of the natural human monoclonal antibody against type 4 adenovirus is shown in SEQ ID NO: 5, and the amino acid sequence of the light chain constant region is shown in SEQ ID NO: 7. The light chain is Kappa chain.

[0010] Secondly, the present application provides a polynucleotide encoding the heavy chain and the light chain of the natural human monoclonal antibody against type 4 adenovirus. The sequence of the polynucleotide encoding the heavy chain variable region of the natural human monoclonal antibody against type 4 adenovirus is shown in SEQ ID NO: 2, and the sequence of the polynucleotide encoding the light chain variable region is shown in SEQ ID NO: 4.

[0011] In a preferred embodiment, the sequence of the polynucleotide encoding the heavy chain constant region of the natural human monoclonal antibody against type 4 adenovirus is shown in SEQ ID NO: 6, and the sequence of the polynucleotide encoding the light chain constant region is shown in SEQ ID NO: 8.

[0012] Thirdly, the present application provides a vector containing the polynucleotide encoding the heavy chain and light chain of the monoclonal antibody. The vectors for expressing the genetically engineered foreign genes in the art, especially the eukaryotic expression vectors, can all be used as the vector of the present application. In a specific embodiment of the present application, the vector is pcDNA3.4.

[0013] Fourthly, the present application provides a host cell containing the vector containing the polynucleotide encoding the heavy chain and light chain of the monoclonal antibody. The host cells for expressing the genetically engineered foreign genes in the art, especially the eukaryotic host cells, can all be used as the host cell of the present application.

[0014] In a preferred embodiment, the cell is a 293T cell or an Expi 293F cell.

[0015] Fifthly, the present application provides the use of the natural human monoclonal antibody against the type 4 adenovirus as described above in the preparation of a medicament for treating and / or preventing the disease caused by the type 4 adenovirus. Since the antibody against the Hexon protein of the type 4 adenovirus is an important neutralizing antibody, the natural human monoclonal antibody against the type 4 adenovirus provided by the present application can be prepared into a medicament for treating or preventing the adenovirus infection.

[0016] Through the antigen-antibody binding epitope analysis of the present application, the binding epitope of the antibody E2DG is located in the amino acid residues 155-266 of the Hexon protein, which is obviously different from the binding epitopes of the antibody B1AG with the heavy chain variable region amino acid sequence and the light chain variable region amino acid sequence as shown in SEQ ID NO. 11 and SEQ ID NO. 12, respectively, and the antibody E3WS with the heavy chain variable region amino acid sequence and the light chain variable region amino acid sequence as shown in SEQ ID NO. 13 and SEQ ID NO. 14, respectively. Therefore, in a specific embodiment of the present application, the present application provides a therapeutic antibody composition for the adenovirus cocktail therapy, which comprises the antibody E2DG with the heavy chain variable region amino acid sequence and the light chain variable region amino acid sequence as shown in SEQ ID NO. 1 and SEQ ID NO. 3, respectively, and the antibody B1AG with the heavy chain variable region amino acid sequence and the light chain variable region amino acid sequence as shown in SEQ ID NO. 11 and SEQ ID NO. 12, respectively, or the antibody E2DG with the heavy chain variable region amino acid sequence and the light chain variable region amino acid sequence as shown in SEQ ID NO. 1 and SEQ ID NO. 3, respectively, and the antibody E3WS with the heavy chain variable region amino acid sequence and the light chain variable region amino acid sequence as shown in SEQ ID NO. 13 and SEQ ID NO. 14, respectively; or the composition comprises the antibody 1QH7, the antibody B1AG and the antibody E3WS.

[0017] Sixthly, the present application provides a detection kit containing the anti-adenovirus type 4 human monoclonal antibody as described above. The kit can be used for diagnosis, monitoring and clinical efficacy evaluation of adenovirus type 4 infection, but not limited to.

[0018] Finally, the present application provides an antibody composition. Through the antigen-antibody binding epitope analysis of the present application, the binding epitope of antibody E2DG is located in the amino acid residue interval of 155-266 of Hexon protein, which is obviously different from the binding epitope of antibody B1AG with the heavy chain variable region amino acid sequence and light chain variable region amino acid sequence as shown in SEQ ID NO. 11 and SEQ ID NO. 12 respectively, and the binding epitope of antibody E3WS with the heavy chain variable region amino acid sequence and light chain variable region amino acid sequence as shown in SEQ ID NO. 13 and SEQ ID NO. 14 respectively. Therefore, in a specific embodiment of the present application, the present application provides an antibody composition for a double antibody sandwich immunoassay of Hexon antigen, which includes two antibodies, one of which is used as a capture antibody and the other as a detection antibody, and the composition includes antibody E2DG with the heavy chain variable region amino acid sequence and light chain variable region amino acid sequence as shown in SEQ ID NO. 1 and SEQ ID NO. 3 respectively, and antibody B1AG with the heavy chain variable region amino acid sequence and light chain variable region amino acid sequence as shown in SEQ ID NO. 11 and SEQ ID NO. 12 respectively, or the composition includes antibody E2DG with the heavy chain variable region amino acid sequence and light chain variable region amino acid sequence as shown in SEQ ID NO. 1 and SEQ ID NO. 3 respectively, and antibody E3WS with the heavy chain variable region amino acid sequence and light chain variable region amino acid sequence as shown in SEQ ID NO. 13 and SEQ ID NO. 14 respectively.

[0019] The present application screens a natural human monoclonal antibody E2DG of adenovirus type 4 from PBMCs samples of convalescent patients infected with adenovirus type 4 by single cell sequencing technology, and the heavy chain variable region and light chain variable region of the antibody have unique CDR regions, which have excellent binding activity with Hexon protein of adenovirus type 4, EC 50 The antibody can inhibit the specific binding of adenovirus type 4 to host cells A549, and the IC 50Reaching 6.124 ng / ml, the excellent neutralizing and inhibitory activity demonstrates the promising application of the antibody provided by this invention as a neutralizing antibody in the preparation of drugs for the treatment and / or prevention of adenovirus type 4 disease. Simultaneously, the antibody's natural human origin ensures the safety of clinical use. Furthermore, the excellent antigen-binding activity and affinity give the antibody provided by this invention promise as a detection antibody for the immunological detection of adenovirus type 4 antigens, such as in the fields of enzyme-linked immunosorbent assay (ELISA), radioimmunoassay, and chemiluminescent immunoassay based on antigen-antibody specific immune reactions. Attached Figure Description

[0020] Figure 1 Visualization of Hexon-specific B cell sorting and single-cell sequencing by flow cytometry; Figure 2 Image showing the results of SDS-PAGE analysis of expressed antibodies; Figure 3 The antibody-antigen binding activity detection diagram of the present invention; Figure 4 The antibody-antigen neutralization activity detection diagram of the present invention; Figure 5 A schematic diagram illustrating the thermal stability detection of the antibody of this invention; Figure 6 A schematic diagram of the competitive binding epitopes of the antibodies of this invention; Figure 7 A schematic diagram illustrating the construction of the AlphaFold3 structure prediction and visualization model for the antigen-antibody complex. Detailed Implementation

[0021] The present invention will be further described below with reference to specific embodiments, and the advantages and features of the present invention will become clearer as a result of the description. However, these embodiments are merely exemplary and do not constitute any limitation on the scope of protection defined by the claims of the present invention.

[0022] In the context of the present application, the term antibody refers to a protein molecule that functions as a receptor to specifically recognize an antigen, and includes an immunoglobulin molecule that is immunoreactive with a specific antigen, including whole antibodies, diabodies, triabodies, and multibodies; bispecific antibodies; chimeric antibodies; recombinant and engineered antibodies, and fragments thereof, as long as the antibody includes an antigen-binding region. The immunoglobulin molecule has a heavy chain and a light chain, each of which includes a constant region and a variable region, wherein the variable region of each of the heavy chain and the light chain is referred to as a variable heavy chain (VH) and a variable light chain (VL), respectively, and the antibody light chain can be classified into two types of a kappa chain and a lambda chain. The variable region of the heavy chain and the light chain includes four framework regions interrupted by three hypervariable regions, also referred to as complementarity determining regions (CDRs), wherein the main role of the CDRs is to bind to an epitope of an antigen.

[0023] Example 1. Screening and preparation of monoclonal antibodies 1. PBMC isolation, obtaining of Hexon-specific B cells, and single-cell library sequencing 20 ml of venous blood was collected from a volunteer infected with adenovirus type 4 in an anticoagulant tube containing heparin. 20 ml of blood sample was used to isolate mononuclear cells (PBMCs) by Ficoll; after cell counting, SONY flow cytometry was used to sort B cells specific to adenovirus type 4 Hexon protein (GENBANK: AAT97450.1) from PBMCs.

[0024] Single B cells were sequenced after isolation using 10x Genomics technology (Chromium, droplet method) (the related kits used are shown in Table 1). Single cells were mixed with single gel beads through oil phase by 10x genomics instrument, and the cell membrane was broken to release the mRNA in the cell after forming water-in-oil small droplets. The free mRNA mixed with the water phase in the small droplets, that is, contacted with reverse transcriptase, nucleic acid primers combined with gel beads, and dNTP substrate, and then reverse transcription reaction (RT: 5'-AAGCAGTGGTATCAACGCAGAGTACTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTVN-3, SEQ ID NO. 15) occurred. All the water phase in the emulsion was extracted, and the reverse transcription product molecules were added with adapters and subjected to PCR amplification (F: 5'-CTACACGACGCTCTTCCGATCT-3, SEQ ID NO. 16; R: 5'-AAGCAGTGGTATCAACGCAGAG-3', SEQ ID NO. 17) to establish an Illumina sequencing library. The library was sequenced using an Illumina sequencer (Read 1 sequencing primer: 5'-ACACTCTTTCCCTACACGACGCTCTTCCGATCT-3', SEQ ID NO. 18; Read 2 sequencing primer 5'-GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT-3', SEQ ID NO. 19), and the data was processed using cell Ranger single cell V8.0.1 software. Then, the RNA-seq alignment software STAT (spliced Transcripts Alignment to Reference) was used to align and analyze the processed Reads and the reference genome (Homo sapiens.GRCh38.dna.primary assembly.fa), and the LoupeBrowser 8 software was used for visual processing.

[0025] Table 1. Kits used for sequencing analysis after isolation of single B cells

[0026]

[0027] Results: As shown in Figure 1 , a total of 1972 specific B cell sequence information was obtained by sequencing, which can be classified into 3 B cell clusters, the blue cell cluster represents Pre B cells, the green cell cluster represents memory B cells, and the orange cell cluster represents plasma cells.

[0028] 2. Expression plasmid construction and antibody preparation Construct the expression plasmid and prepare the monoclonal antibody. The method is as follows: 1) Homologous recombination of the antibody variable region obtained by single cell sequencing into pcDNA3.4 (Shengong Biology), transform into TOP10 competent cells, and then extract the plasmid; 2) Cell inoculation: inoculate 1 ml of 293T cells with a density of 2x10 5 cells / ml into a 24-well cell culture plate and incubate at 37°C for 24 hours; 3) Cell co-transfection: 24 hours later, add the successfully constructed heavy chain and light chain plasmids to 2 μl of transfection reagent Turbofect (Thermo Scientific, R0531), incubate for 15-20 minutes, and then add dropwise to the 293T cell culture well of the overnight culture. Incubate at 37°C in a cell incubator containing 5% CO2 for 48 hours, then collect the cell culture supernatant for use.

[0029] 3. ELISA screening of antibodies with binding activity 1) One day before the experiment, coat 2 μg / ml of Hexon protein (Genebank No. AY594253.1 self-constructed) in a 96-well ELISA plate, 100 μl per well, and incubate overnight at 4°C; 2) Discard the coating solution, add 100 μl of blocking solution to each well, and incubate at 37°C for 1 hour; 3) Discard the blocking solution, add 100 μl of transfection cell culture supernatant, and incubate at 37°C for 1 hour; 4) Discard the supernatant, dilute the HRP-labeled goat anti-human IgG secondary antibody (Abcam, ab97225) with a dilution of 1:10,000, and add 100 μl to each well of the ELISA plate, incubate at 37°C for 1 hour; 5) Discard the supernatant, add 100 μl of TMB single-component color developing solution to each well, develop at room temperature in the dark for 6 minutes, then add 50 μl of stop solution to each well to stop the reaction; 6) Detect the OD value at 450-630 nm using an enzyme marker, and save the raw data.

[0030] 4. Large-scale preparation of antibodies Co-transfect Expi293F cells with expression vectors of antibody E2DG heavy chain and light chain with neutralizing activity, change fresh medium 6-8h after transfection, and culture in 37℃ 8% CO2 incubator for 96h. Collect the transfection supernatant, centrifuge at 4000rpm for 10 minutes, and purify by protein A affinity chromatography. Use SDS-PAGE to test the expression and purification of the antibody.

[0031] 5. Results The results of antibody E2DG are shown in Figure 2 , and the target antibody is confirmed by SDS-PAGE method. The antibody heavy chain (55kDa, reduced state) and light chain (25kDa, reduced state) after dissociation can be clearly observed, and the molecular weight is 150kDa (non-reduced state), which is consistent with the expected value (marker 26616).

[0032] Example 2. ELISA detection of antibody binding activity 1) One day before the experiment, 96-well enzyme-linked plates were coated with 2 µg / ml of Hexon protein, 100 µl of coating solution per well, 4℃ overnight; 2) Discard the coating solution, add 100 µl of blocking solution per well, and incubate at 37°C for 1 hour; 3) Discard the blocking solution, add 1 µg / ml of antibody E2DG, dilute 1:3, and incubate at 37°C for 1 hour; 4) Discard the primary antibody, dilute the HRP-labeled goat anti-human IgG secondary antibody (Abeam, ab97225) at 1:10000, and add 100 µl to the corresponding wells of the ELISA plate, incubate at 37°C for 1 hour; 5) Discard the secondary antibody, add 100 µl of TMB single-component color developing solution to each well, develop for 6 minutes at room temperature in the dark, and then add 50 µl of stop solution to each well to stop the reaction; 6) Detect the OD value at 450-630nm with a microplate reader, and save the raw data.

[0033] The results are shown in Figure 3 , and the EC 50 of antibody E2DG reaches 30.94 ng / ml.

[0034] Example 3. Detection of antibody neutralizing activity The antibody neutralization level was determined by HAdv-4 luciferase-containing pseudovirus (purchased from Promega, catalog number E1501). The specific steps are as follows: 1) 96-well plate, 50 μl of antibody E2DG at a concentration of 100 μg / ml, diluted at a gradient of 1:3, and an equal volume of HAdV-4 luciferase, 37°C incubation for 1 hour; 2) 1 hour later, 2*10 5 cells / ml of A549 cells 100 μl were added for 37°C incubation for 24 hours; 3) 24 hours later, 100 μl of lysis solution was added for reading; The results, as shown in Figure 4 , showed that the IC 50 of antibody E2DG reached 6.124 ng / ml.

[0035] Example 4. DSC detection of antibody thermal stability Differential scanning calorimetry (DSC) studies the transition or process caused by temperature change, most commonly the "thermal melting" caused by temperature rise, so DSC is used to study the thermal stability of antibody 1QH7. The determination method is as follows: 1. Start the instrument and DSC-RUN software in advance, preheat for 1 h; 2. Connect the cleaning device, 800 ml of washing solution (5% decon90) and 1.5 L of pure water; 3. Open the 1.5 ml centrifuge tube containing PBS and the antibody to be tested in the degassing device, degas at high pressure 635 Hg for 10 min, and wait for use; 4. After the instrument is cleaned, use a pipette to slowly add 800 μl of PBS and the antibody sample to be tested to the reference hole and the sample hole, respectively, and repeatedly blow to remove air bubbles in the holes. A black plastic cap is used to tighten the reference hole and the sample hole, respectively, and the instrument cover is tightened; 5. Observe whether the instrument thermal value is stable, and then start to pressurize to 3 atm when it is stable; 6. After the pressurization is completed, set the temperature to 25-100°C, balance for 600 s, and run the program; 7. After the program runs to 100°C, the system automatically cools down, the program can be terminated, and the temperature is cooled to about 30°C, the pressure is reduced to 0 atm, the instrument cover is opened, and the sample is sucked out, and the instrument is cleaned.

[0036] The results, as shown in Figure 5 , showed that the melting temperature of the Fab region of antibody E2DG was 67.52°C, and it had good thermal stability.

[0037] Example 5. Antibody binding epitope competition The natural human neutralizing antibodies of human adenovirus type 4 are labeled by biotin labeling method, and the binding epitopes of the antibodies are analyzed in depth by competitive ELISA. In addition to the antibody E2DG, the antibody 1QH7 whose heavy chain variable region amino acid sequence and light chain variable region amino acid sequence are respectively shown as SEQ ID NO. 9 and SEQ ID NO. 10, the antibody B1AG whose heavy chain variable region amino acid sequence and light chain variable region amino acid sequence are respectively shown as SEQ ID NO. 11 and SEQ ID NO. 12, and the antibody E3WS whose heavy chain variable region amino acid sequence and light chain variable region amino acid sequence are respectively shown as SEQ ID NO. 13 and SEQ ID NO. 14 are involved in the experiment.

[0038] 1. Antibody binding epitope competition experiment The determination method is as follows: (1) 2 days before the experiment, 200 μg of antibodies are labeled with EZ-Link™ Sulfo-NHS-Biotin (purchased from Thermo Scientific, item number 21338) at a molar ratio of 1:5 (antibody: biotin) overnight in the dark at 4 degrees, the sample is irradiated with strong light to remove free biotin for 24 h, and the free biotin is removed by centrifugation multiple times using a desalting column (purchased from Thermo Scientific, item number 89882); (2) 96-well enzyme-labeled plates are coated with 100 μL of Hexon antigen protein at a concentration of 2 μg / mL, and incubated overnight at 4 degrees; (3) On the day of the experiment, the enzyme-labeled plates are washed 3 times with a BioTek 405 LS plate washer, 100 μL of blocking solution is added to each well, and incubated at 37 degrees for 1 h; (4) Wash the plate 3 times, add 50 μL of un-labeled biotin antibody to each well, 20 μg / mL, add in a matrix horizontally, and incubate at 37 degrees for 1 h; (5) Add 50 μL of labeled biotin antibody to each well, 1 μg / mL, add in a matrix vertically, and incubate at 37 degrees for 1 h, the order is the same as that of the blocking antibody; (6) Wash the plate 3 times, add 100 μL of Streptavidin-HRP (Abcam, ab7403) antibody diluted 1:10,000, and incubate at 37 degrees for 1 h; (7) Wash the plate 3 times, add 100 μL of TMB single-component color developing liquid, and develop color at room temperature for 6 min; (8) Add 50 μL of stop solution, read OD value at 450-630 nm wavelength, competition value = (biotinylated antibody OD value in the presence of competing antibody / biotinylated antibody OD value in the presence of irrelevant antibody) x 100%, competition value < 33.3, determined as strong competition; > 66.7, determined as no competition; between 33.3 and 66.7, determined as weak competition.

[0039] The results are shown in Table 1. Figure 6 As shown in Table 1, antibody E2DG has no epitope overlap with other neutralizing antibodies except for antibody 1QH7 which has overlapping binding epitopes.

[0040] 2. Antigen-antibody complex AlphaFold 3 structure prediction and visualization model construction The amino acid sequence of the adenovirus antigen Hexon protein was downloaded from the GENBANK gene sequence database (https: / / www.ncbi.nlm.nih.gov / genbank / ). The heavy chain and light chain sequences of the monoclonal antibodies screened and the Hexon antigen sequence were input into the locally deployed AlphaFold 3.0.0 to predict the antigen-antibody complex structure of the antibodies and the Hexon protein. In the structure prediction process, the default model parameters were used, and the confidence indicators such as pLDDT, PAE, pTM, and ipTM were integrated to select the model with the highest ranking_score for subsequent analysis. On this basis, the "Structure Analysis / Matcher" tool in ChimeraX 1.2.4 software was used to superimpose the prediction models of multiple neutralizing antibodies with the Hexon protein template structure, and finally a visualization model of the antibody-Hexon protein complex was simulated.

[0041] The results are shown in Table 1. Figure 7 As shown in Table 1, antibody E2DG has no epitope overlap with other neutralizing antibodies except for antibody 1QH7 which has overlapping binding epitopes.

[0042] The results of the analysis show that antibody E2DG has application prospects for immunodetection of the Hexon antigen in a double antibody sandwich method in combination with antibody B1AG or antibody E3WS, as well as application prospects for a cocktail therapy in combination with antibody B1AG and / or antibody E3WS.

Claims

1. A human monoclonal antibody against adenovirus type 4, characterized in that, The amino acid sequences of the CDR1, CDR2 and CDR3 regions of the heavy chain variable region of the anti-adenovirus type 4 human monoclonal antibody are shown in SEQ ID NO: 1 at positions 26-33, 51-58 and 97-116, respectively; and the amino acid sequences of the CDR1, CDR2 and CDR3 regions of the light chain variable region are shown in SEQ ID NO: 3 at positions 26-36, 54-56 and 93-101, respectively.

2. The human monoclonal antibody against type 4 adenovirus according to claim 1, characterized in that, The amino acid sequence of the heavy chain variable region of the anti-adenovirus type 4 human monoclonal antibody is shown in SEQ ID NO: 1, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO:

3.

3. The human monoclonal antibody against adenovirus type 4 according to claim 2, characterized in that, The amino acid sequence of the heavy chain constant region of the anti-adenovirus type 4 human monoclonal antibody is shown in SEQ ID NO: 5, and the amino acid sequence of the light chain constant region is shown in SEQ ID NO:

7.

4. A polynucleotide encoding the heavy and light chains of the human monoclonal antibody against human adenovirus type 4 according to any one of claims 1 to 3, characterized in that, The sequence of the polynucleotide encoding the heavy chain variable region of the anti-adenovirus type 4 human monoclonal antibody is shown in SEQ ID NO: 2, and the sequence of the polynucleotide encoding the light chain variable region of the anti-adenovirus type 4 human monoclonal antibody is shown in SEQ ID NO:

4.

5. The polynucleotide of claim 4, wherein, The sequence of the polynucleotide encoding the heavy chain constant region of the anti-adenovirus type 4 human monoclonal antibody is shown in SEQ ID NO: 6, and the sequence of the polynucleotide encoding the light chain constant region of the anti-adenovirus type 4 human monoclonal antibody is shown in SEQ ID NO:

8.

6. A vector containing the polynucleotide encoding the heavy chain and the light chain of the anti-adenovirus type 4 human monoclonal antibody of claim 5.

7. Use of the anti-adenovirus type 4 human monoclonal antibody of any one of claims 1-3 in the preparation of a medicament for treating and / or preventing adenovirus type 4 disease.

8. A detection kit containing the anti-adenovirus type 4 human monoclonal antibody of any one of claims 1-3.

9. An antibody composition, characterized in that, The antibody composition comprises the antibody whose heavy chain variable region amino acid sequence and light chain variable region amino acid sequence are shown in SEQ ID NO. 1 and SEQ ID NO. 3, respectively, and the antibody whose heavy chain variable region amino acid sequence and light chain variable region amino acid sequence are shown in SEQ ID NO. 11 and SEQ ID NO. 12, respectively, or The composition comprises the antibody whose heavy chain variable region amino acid sequence and light chain variable region amino acid sequence are shown in SEQ ID NO. 1 and SEQ ID NO. 3, respectively, and the antibody whose heavy chain variable region amino acid sequence and light chain variable region amino acid sequence are shown in SEQ ID NO. 13 and SEQ ID NO. 14, respectively.

10. The antibody composition of claim 9, wherein, The antibody composition includes an antibody having a heavy chain variable region amino acid sequence and a light chain variable region amino acid sequence as set forth in SEQ ID NO. 1 and SEQ ID NO. 3, respectively, an antibody having a heavy chain variable region amino acid sequence and a light chain variable region amino acid sequence as set forth in SEQ ID NO. 11 and SEQ ID NO. 12, respectively, and an antibody having a heavy chain variable region amino acid sequence and a light chain variable region amino acid sequence as set forth in SEQ ID NO. 13 and SEQ ID NO. 14, respectively. The antibody composition includes an antibody having a heavy chain variable region amino acid sequence and a light chain variable region amino acid sequence as set forth in SEQ ID NO. 1 and SEQ ID NO. 3, respectively, an antibody having a heavy chain variable region amino acid sequence and a light chain variable region amino acid sequence as set forth in SEQ ID NO. 11 and SEQ ID NO. 12, respectively, and an antibody having a heavy chain variable region amino acid sequence and a light chain variable region amino acid sequence as set forth in SEQ ID NO. 13 and SEQ ID NO. 14, respectively.

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