Chikungunya virus monoclonal antibody and application thereof in preparation of preventive and therapeutic antibody drugs
By preparing a rabbit monoclonal antibody targeting the CHIKV E2 protein, the problem of the lack of effective drugs for CHIKV infection in the existing technology has been solved, and efficient in vitro neutralization and in vivo protection effects have been achieved.
Patent Information
- Application Number
- CN202610016258.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2026-01-07
- Publication Date
- 2026-02-10
AI Technical Summary
There is a lack of effective drugs for treating chikungunya virus (CHIKV) infection in the current technology, especially the efficacy of neutralizing antibodies against the CHIKV E2 protein in non-human primates has been poorly evaluated, and the neutralizing activity of existing monoclonal antibodies is not strong enough.
Rabbit monoclonal antibodies against the CHIKV E2 protein were prepared, and their amino acid sequence and in vitro and in vivo applications were provided. These antibodies are developed as preventive or therapeutic antibodies by efficiently neutralizing CHIKV infection.
This antibody can block CHIKV infection of target cells in vitro and protect mice against lethal infection in vivo, demonstrating highly efficient neutralizing activity and protective effect.
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Abstract
Description
TECHNICAL FIELD
[0001] The present application relates to the technical field of biological medicine engineering, and particularly relates to an amino acid sequence of a Chikungunya virus envelope E2 protein monoclonal antibody and application of the amino acid sequence in development of a therapeutic or prophylactic antibody against Chikungunya virus infection. BACKGROUND
[0002] Chikungunya virus (CHIKV) is the pathogen causing Chikungunya fever, and belongs to the Togaviridae family and Alphavirus genus. The genome of CHIKV is a single-stranded positive RNA. CHIKV is an important arbovirus, and Aedes mosquitoes are the main transmission media. Human beings are generally susceptible to CHIKV. In addition to causing fever and rash, CHIKV can also cause chronic arthritis lasting more than half a year. In recent years, it has been reported that CHIKV can cause more serious diseases, including encephalitis and hemorrhagic diseases. At present, CHIKV has spread to more than 100 countries and regions in the world, and 39% of the global population is at risk of CHIKV infection, causing millions of infections every year, which is a major burden on global public health. At present, there is no effective treatment for CHIKV infection.
[0003] The CHIKV genome encodes two open reading frames (ORF). The 5' end ORF encodes non-structural proteins (nsp1, nsp2, nsp3, nsp4), which are mainly responsible for the replication of genomic RNA; and the 3' end ORF encodes structural proteins (capsid protein C, envelope protein E3, envelope protein E2, envelope protein E1), which are responsible for the formation of virus particles.
[0004] The translation product of the structural protein is a polyprotein precursor. The capsid protein at the amino terminal of the polyprotein precursor is released from the polyprotein precursor by the protease hydrolysis function of the capsid protein, and the carboxy terminal polypeptide is cut by a host signal peptide in the endoplasmic reticulum to form a precursor pE2 protein, a 6K protein and an E1 protein. Subsequently, the pE2 is cut by a host cell furin in the Golgi apparatus to form E3 and E2 proteins.
[0005] The capsid protein contains two domains: an N-terminal RNA-binding domain and a C-terminal protease domain. The RNA-binding domain is rich in positively charged lysine and arginine, which interacts with the negatively charged genomic RNA. The envelope protein E2 is the receptor-binding protein of the virus, mainly responsible for the binding of CHIKV to the cell surface receptor. The E2 protein is structurally divided into four parts: the extramembrane region, the stem region, the transmembrane region, and the intramembrane region. The full-length CHIKV E2 protein includes 423 amino acid residues (aa), of which the extracellular region is about 260 aa, the stem region is about 100 aa, the transmembrane region is about 30 aa, and the intramembrane region is about 33 aa. The extramembrane region plays a key role in virus adsorption to host cells and transmembrane infection, and contains epitopes of neutralizing antibodies; the transmembrane region enters the lipid bilayer, and the intramembrane region interacts with the nucleocapsid protein inside the virus particle. The E1 protein belongs to the second class of fusion proteins and is mainly responsible for membrane fusion. E1 is also structurally divided into four parts: the extramembrane region, the stem region, the transmembrane region, and the intramembrane region. The E2 protein and the E2 protein form a heterodimer and are anchored on the envelope surface of the virus particle.
[0006] CHIKV infection of cells begins with the binding of the virus to the cell surface receptor. After the E2 protein of CHIKV binds to the receptor, the virus particle forms an endosome through receptor-mediated endocytosis, enters the cell, and as the endosome matures, the acidification of the endosome causes a change in the conformation of the CHIKV E2-E1 heterodimer, leading to the exposure and insertion of the fusion peptide in the E1 protein into the endosome membrane, inducing fusion of the virus envelope with the endosome membrane. After membrane fusion occurs, the viral nucleocapsid structure is released into the cytoplasm and rapidly disintegrates within a short period of time, releasing the viral genomic RNA and initiating viral gene replication and protein expression.
[0007] Antibody therapy is an effective method for treating severe viral infections and has shown good results in treating viral infections such as Ebola virus and respiratory syncytial virus. Monoclonal antibodies have become a new class of specific drugs for treating viral infections and are currently a hot topic in the development of antiviral drugs. Specific monoclonal antibodies against CHIKV have received widespread attention as a potential new treatment. Although some monoclonal antibodies against CHIKV have been reported, the neutralizing activity of the antibodies is not strong enough, and few monoclonal antibodies have been evaluated for efficacy in non-human primates. Therefore, the development of new antibodies for the prevention and treatment of CHIKV is of urgent and practical significance in filling the gap in specific drugs for CHIKV.
[0008] Compared with mouse monoclonal antibodies, rabbit monoclonal antibodies have a series of unique advantages in molecular structure, immune mechanism and engineering: 1. wider immune recognition spectrum: the B cell library capacity of rabbits is larger, which can produce a wider and more diverse antibody spectrum, which is conducive to recognizing more types of antigen epitopes; 2. higher affinity and specificity: the antibodies are affinity matured through high-frequency somatic hypermutation and unique gene conversion double mechanism, and the complementarity determining region structure is unique, which together makes them usually exhibit higher affinity and specificity to antigens; 3. stronger structural stability: rabbit antibodies contain an additional disulfide bond in the heavy chain variable region, and there is also a connecting disulfide bond between the variable region and the constant region. This unique disulfide bond network significantly enhances the structural stability of the molecule; 4. more convenient humanization modification: rabbit monoclonal antibodies are easy to be humanized, and there are many humanized rabbit monoclonal antibody drugs on the market. SUMMARY
[0009] CHIKV E2 protein is a key protein for mediating viral invasion of host cells and a main protein for inducing neutralizing antibodies. The present application takes CHIKV E2 protein as a target antigen to prepare rabbit monoclonal antibodies, and obtains a neutralizing antibody that can efficiently and broadly neutralize major epidemic genotypes of CHIKV worldwide, which can be further developed as a prophylactic or therapeutic antibody against CHIKV infection.
[0010] Specifically, the present application provides a CHIKV E2 protein rabbit monoclonal antibody and its use. The monoclonal antibody can block the infection of CHIKV to target cells in vitro, and can protect mice against lethal infection of CHIKV in vivo. The deoxynucleotide sequence of the antibody heavy chain signal peptide and variable region gene is shown as SEQ ID NO: 1, and the amino acid sequence of the antibody heavy chain signal peptide and variable region protein is shown as SEQ ID NO: 2. The deoxynucleotide sequence of the antibody light chain signal peptide and variable region gene is shown as SEQ ID NO: 3, and the amino acid sequence of the antibody light chain signal peptide and variable region protein is shown as SEQ ID NO: 4.
[0011] More specifically, the present application provides a CHIKV envelope protein E2 monoclonal antibody, characterized in that the amino acid sequence of the antibody includes the antibody heavy chain variable region and the antibody light chain variable region. The amino acid sequence of the antibody heavy chain variable region is shown as SEQ ID NO: 2. The amino acid sequence of the antibody light chain variable region is shown as SEQ ID NO: 4.
[0012] Further, the present application provides a CHIKV envelope protein E2 monoclonal antibody, which is further characterized in that: The deoxynucleotide sequence of the antibody heavy chain variable region gene is shown as SEQ ID NO: 1.
[0013] Further, the present application provides a CHIKV envelope protein E2 monoclonal antibody, which is characterized in that: The deoxynucleotide sequence of the antibody light chain variable region gene is shown as SEQ ID NO: 3.
[0014] Further, the present application provides a CHIKV envelope protein E2 monoclonal antibody, which is characterized in that: The antibody as a therapeutic or prophylactic component, a medicament, or for preparing a medicament; The therapeutic component / medicament can block the infection of CHIKV to target cells in vitro.
[0015] Further, the present application provides a CHIKV envelope protein E2 rabbit monoclonal antibody, which is characterized in that: The antibody as a therapeutic or prophylactic component, a medicament, or for preparing a medicament; The therapeutic component / medicament can protect mice against lethal infection of CHIKV in vivo.
[0016] That is, simply speaking: the antibody can block the infection of CHIKV to target cells in vitro.
[0017] The antibody can protect mice against lethal infection of CHIKV in vivo.
[0018] Further, the present application provides a CHIKV envelope protein E2 rabbit monoclonal antibody, which is characterized in that: Application in developing therapeutic or prophylactic antibodies against CHIKV infection. BRIEF DESCRIPTION OF DRAWINGS
[0019] Figure 1 ELISA detection of the binding of monoclonal antibody 6E7C4 to CHIKV envelope protein E2 Figure 2 Infection neutralization of CHIKV 37997 strain by monoclonal antibody 6E7C4 Figure 3 Protection of mice against CHIKV Ross strain attack by monoclonal antibody 6E7C4 (weight change graph) Figure 4 Protection of mice against CHIKV Ross strain attack by monoclonal antibody 6E7C4 (survival rate graph) DETAILED DESCRIPTION
[0020] The application will be described in detail below with reference to the embodiments and drawings of the application. The following embodiments are implemented on the premise of the technical solutions of the application, and specific implementation modes and operation processes are given. However, the protection scope of the application is not limited to the following embodiments. All operations related to CHIKV in the application are performed in a biosafety level 3 laboratory of the Navy Medical University of the Chinese People's Liberation Army.
[0021] I. Main experimental materials 1. CHIKV E2 protein, E2 protein of the Indian Ocean lineage LR2006 strain of the East / Central / Southern African type CHIKV recombinantly expressed by human embryonic kidney 293T cells, and grown in serum-free medium by freestyle 293F cells of the Biomedical Defense Research Lab of the Navy Medical University of the Chinese People's Liberation Army.
[0022] 2. CHIKV virus, including the 37997 strain of the Asian type and the Ross strain of the East / Central / Southern African type, synthesized by the Biomedical Defense Research Lab of the Navy Medical University of the Chinese People's Liberation Army using reverse genetics technology.
[0023] 3. C57BL / 6 mice purchased from Shanghai Xipu Bikey Experimental Animal Co., Ltd.
[0024] 4. Mouse anti-CHIKV polyclonal antibody prepared by the Biomedical Defense Research Lab of the Navy Medical University of the Chinese People's Liberation Army by immunizing mice with b-propiolactone inactivated Ross strain CHIKV.
[0025] II. Experimental methods and results 1. Preparation and identification process of rabbit monoclonal antibody The recombinantly expressed CHIKV E2 protein was used to immunize New Zealand rabbits in combination with complete Freund's adjuvant. 0.2 mg of CHIKV E2 protein was dissolved in phosphate buffered saline (PBS) to a final volume of 0.5 ml, and was thoroughly emulsified with 0.5 ml of complete Freund's adjuvant. The New Zealand rabbits were immunized by subcutaneous injection at multiple sites. Booster immunizations were performed once at 3, 6, and 9 weeks after the initial immunization, for a total of 3 times. The amount of CHIKV E2 protein used for the booster immunization was 0.1 mg, and the adjuvant used was incomplete Freund's adjuvant. The rabbits were subcutaneously immunized. Three weeks after each immunization, blood was collected from the rabbit ear marginal vein, and the serum was tested for IgG antibodies by ELISA (as described below). After 12 weeks, the rabbits were anesthetized with isoflurane ether, and blood was collected by cardiac puncture. After the rabbits were sacrificed, the spleen was removed, cut into small pieces, ground on a nylon mesh, and filtered to prepare a single cell suspension. The single spleen cells expressing CHIKV E2 protein IgG antibodies were sorted by flow cytometry, and were inoculated into a 96-well plate. The cells were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum and 100 U / ml penicillin and 100 mg / ml streptomycin. The 96-well plate was placed in a 37°C cell culture incubator with 5% CO2 and saturated humidity for about 7-10 days. When the cells had grown to cover the bottom of the plate, the culture supernatant was tested for the presence of CHIKV E2 IgG antibodies by ELISA. The total RNA of the antibody-positive cells was extracted, and the antibody heavy chain signal peptide and variable region gene (Signal-VH) and the light chain signal peptide and variable region gene (Signal-VL) were amplified by reverse transcription polymerase chain reaction. The PCR products were recovered by agarose gel electrophoresis, inserted into a pMD18T vector (Takara), and subjected to DNA sequencing. After the antibody heavy chain Signal-VH and the rabbit IgG1 CH1-CH2-CH3 were spliced into a complete heavy chain gene, and the light chain Signal-VL gene and the rabbit light chain constant region gene were spliced into a complete light chain gene. The full-length heavy chain and light chain genes were inserted into the cloning sites of the mammalian cell expression vector pcDNA3.4 to construct expression plasmids. The antibody heavy chain and light chain expression plasmids were co-transfected into 293T cells at a mass ratio of 1:2, and the culture supernatant was tested for the presence of CHIKV E2 protein IgG antibodies by ELISA. The IgG antibody-positive supernatant was subjected to a virus microneutralization experiment to determine whether it had neutralizing activity. If the monoclonal antibody had virus neutralizing activity, the freestyle 293F cells were suspended in serum-free medium and cultured to grow to a volume of 300 ml (1 L of culture flask). The cell density was adjusted to 1 x 10 6When the concentration of the antibody reached 10 μg / ml, the antibody heavy chain and light chain expression plasmids were co-transfected into freestyle 293F cells using a polyethylenimine (PEI) reagent, and the cells were cultured for 72 hours. The antibody was purified from the culture supernatant by Staphylococcus aureus Protein A affinity chromatography, and ELISA, virus neutralization test and mouse challenge protection test were performed.
[0026] 2. ELISA detection of CHIKV E2 protein IgG antibodies CHIKV E2 protein was diluted with 50 mM carbonate coating solution (pH 9.6) to a concentration of 0.5 μg / ml, and 0.1 ml was added to each well of an enzyme-labeled plate containing 0.05 mg of CHIKV E2 protein. The plate was placed in a refrigerator at 4°C overnight. The next day, the coating solution was removed, and each well was washed once with 0.2 ml of PBS buffer. Then, 0.2 ml of PBS buffer containing 3% bovine serum albumin, 5% goat serum and 0.05% Tween 20 (blocking solution, pH 7.4) was added to each well, and the plate was incubated at room temperature for 2 hours. After removing the blocking solution, the wells were washed 3 times with PBS buffer containing 0.05% Tween 20 (washing solution). Then, 0.1 ml of rabbit spleen cell culture supernatant, culture supernatant of 293T cells co-transfected with antibody expression plasmids, or purified monoclonal antibody obtained from culture supernatant of freestyle 293F cells co-transfected with antibody expression plasmids was added to each well, and the plate was placed in a slow shaking horizontal shaker at room temperature for 30 minutes. Then, the reaction solution in each well was discarded, and the wells were washed 5 times with washing solution. Then, 0.1 ml of horseradish peroxidase-labeled anti-rabbit IgG (H+L) (Thermo product) diluted 1:2000 with blocking solution was added to each well, and the plate was placed in a slow shaking horizontal shaker at room temperature for 30 minutes. Then, the enzyme-labeled antibody dilution solution in each well was discarded, and the wells were washed 5 times with washing solution. Then, 0.1 ml of substrate solution containing 5,5'-tetramethylbenzidine (TMB) was added to each well, and the plate was placed in the dark at room temperature for 5 minutes. Then, 50 ml of 2 M sulfuric acid was added, and the plate was mixed well. The optical absorbance value at 450 nm was detected using an enzyme-labeled instrument, and the reference wavelength was 630 nm.
[0027] Figure 1The ELISA light absorption values (A450-630) obtained from different concentrations of a CHIKV E2 protein monoclonal antibody (No. 6E7C4) identified by the display screening are shown in the figure, with rabbit IgG1 (Cell signaling company) as a negative control (Control), and it can be seen that the monoclonal antibody 6E7C4 can specifically bind to the CHIKV E2 protein. The deoxyribonucleotide sequence of the heavy chain variable region gene of the antibody is shown in SEQ ID NO: 1, and the amino acid sequence of the antibody heavy chain variable region protein deduced from the gene sequence is shown in SEQ ID NO: 2. The deoxyribonucleotide sequence of the antibody light chain variable region gene is shown in SEQ ID NO: 3, and the amino acid sequence of the antibody light chain variable region protein deduced from the gene sequence is shown in SEQ ID NO: 4. Further in vitro neutralization test and in vivo mouse challenge protection test were carried out on the monoclonal antibody to evaluate its anti-virus effect.
[0028] Specifically: SEQ ID NO: 1: Deoxyribonucleotide sequence of the signal peptide and variable region of the monoclonal antibody 6E7C4 (1-60 is the deoxyribonucleotide sequence of the signal peptide, and 61-405 is the deoxyribonucleotide sequence of the variable region) 1 ATGGAGACTG GGCTGCGCTG GCTTCTCCTG GTCGCTGTGC TCAAAGGTGT CCAGTGTCGG 61 TCGGTGGAGG AGTCCGGGGG TCGCCTGGTC ACGCCTGGGA CACCCCTGAC ACTCACCTGC 121 ACAGTCTCTG GATTCTCCCT CGATGTCTAT GCAGTGAGTT GGGTCCGCCA GGCTCCAGGG 181 AAGGGTCTGG AATGGATCGG AATCATTTAT GGGACTCCTA ACACAGCCTA CGCGAACTGG 241 GCGAAAGGCC GATTCACCAT CTCCAAAACC TCGACCACGG TGGATTTGAA AATGACCAGT 301 CCGACAACCG AGGACACGGC CACCTATTTC TGTGCCAGAG GGCCGGGTGA TAGTGGAGTT 361 GAATATAATT TGTGGGGCCA AGGCACCCTG GTCACCGTCT CCTCG SEQ ID NO: 2: Amino acid sequence of the heavy chain signal peptide and variable region of mAb 6E7C4 (1-20 is the signal peptide amino acid sequence, 21-135 is the variable region amino acid sequence) 1 METGLRWLLL VAVLKGVQCR SVEESGGRLV TPGTPLTLTC TVSGFSLDVY AVSWVRQAPG 61 KGLEWIGIIY GTPNTAYANW AKGRFTISKT STTVDLKMTS PTTEDTATYF CARGPGDSGV 121 EYNLWGQGTL VTVSS SEQ ID NO: 3: Deoxynucleotide sequence of the light chain signal peptide and variable region of mAb 6E7C4 (1-69 is the signal peptide deoxynucleotide sequence, 70-396 is the variable region deoxynucleotide sequence) 1 ATGGACACGA GGGCCCCCAC TCAGCTGCTG GGGCTCCTGC TGCTCTGGCT CCCAGGTGCC 61 AGATGTGCCT ATGATATGAC CCAGACTCCA GCCTCCGTGG AGGCAGCTGT GGGAGGCACA 121 GTCACCATCA AGTGCCAGGC CAGTCAGAGC ATCAGCAACT ACTTAGCCTG GTATCAGCAG 181 AAACCAGGGC AGCCTCCCAA GCTCCTGATC TATGAAGCAT CCAAACTGGC AACTGGGGTC 241 TCATCGCGGT TCAAAGGCAG TGGATCTGGG ACAGAGTTCA CTCTCACCAT CAACGACCTG 301 GAGTGTGCCG ATGCTGCCGC TTACTACTGT CAACAGGGTT ATAGTGATAC TAATGTTGAT 361 AATGTTTTCG GCGGAGGGAC CGAAGTGGTG GTCAAG SEQ ID NO: 4: Amino acid sequence of the light chain signal peptide and variable region of mAb 6E7C4 (1-23 is the signal peptide amino acid sequence, 24-132 is the variable region amino acid sequence) 1 MDTRAPTQLL GLLLLWLPGA RCAYDMTQTP ASVEAAVGGT VTIKCQASQS ISNYLAWYQQ 61 KPGQPPKLLI YEASKLATGV SSRFKGSGSG TEFTLTINDL ECADAAAYYC QQGYSDTNVD 121 NVFGGGTEVV VK 3. Neutralization activity of mAbs against CHIKV The cultured baby hamster kidney (BHK) cells were subcultured in 96-well plates with 10,000 cells per well in DMEM medium (hereinafter referred to as complete DMEM medium) supplemented with 10% fetal bovine serum and 100 U / ml penicillin and 100 mg / ml streptomycin, in a 37°C cell culture incubator containing 5% CO2and saturated humidity for 12 hours.
[0029] The rabbit monoclonal antibody 6E7C4 was gradiently diluted in a 96-well plate (diluent: complete DMEM medium), with rabbit IgG1 (Cell signaling company) as a negative control, each concentration gradient was aspirated into a 96-well plate, 50 ml per well, 50 ml of diluted CHIKV virus liquid was added to each well, containing about 500 focus forming units (PFUs) of CHIKV, the diluent was complete DMEM medium, mixed well, and placed in a 37°C cell incubator for 30 minutes. Then the liquid in the well was transferred to the BHK cell culture well (the original cell culture medium was first aspirated), and the cells were further cultured for 18 hours. The expression of CHIKV protein in the cells was detected by immunofluorescence. The specific operation is as follows: the culture medium in the culture plate was aspirated, 0.1 ml of methanol was added to each well, the culture plate was placed in a-20°C refrigerator for 20 minutes, then the culture plate was taken out, the methanol was aspirated, each well was washed with phosphate buffered saline (PBS) once, then 0.1 ml of 3% bovine serum albumin (BSA)-PBS containing 3% BSA was added, and the culture plate was slowly shaken on a horizontal shaker at room temperature for 1 hour. The 3% BSA-PBS in the culture plate was aspirated, 0.1 ml of 1% BSA-PBS containing anti-CHIKV polyclonal antibody was added to each well (the antibody was diluted 500 times), and the culture plate was slowly shaken at room temperature for 1 hour. The CHIKV polyclonal antibody working solution in the culture plate was aspirated, each well was washed with PBS for 3 times, then 100 ml of 1% BSA-PBS containing fluorescein Alexa Fluor 488-labeled anti-mouse IgG was added (the fluorescein antibody was diluted 1500 times), and the culture plate was slowly shaken at room temperature for 1 hour. The fluorescein antibody working solution in the culture plate was aspirated, 0.1 ml of DAPI cell nucleus staining solution was added to each well, and the culture plate was slowly shaken at room temperature for 10 minutes. The DAPI cell nucleus staining solution in the culture plate was aspirated, each well was washed with PBS for 3 times, and the green fluorescent positive cells in each well were counted using a cell imaging and analysis system (BioTek Cytation 5 Imaging Reader). Then the neutralization percentage (%) was calculated as follows: the number of positive cells in the monoclonal antibody treated well or the control rabbit IgG treated well / the number of positive cells in the well without antibody.
[0030] From Figure 2 It can be seen that the 6E7C4 monoclonal antibody can efficiently neutralize the infection of the Asian strain 37997 CHIKV on the target cells BHK.
[0031] 4. Evaluation of the anti-viral activity of the monoclonal antibody in vivo using a mouse virus challenge model 8-week-old female C57BL / 6 mice, a total of 20, were randomly divided into 2 groups, 10 in each group, and were infected with CHIKV East / Central / South African Ross strain by nasal instillation, with a volume of 50 ml, containing 1´10 6PFUs of CHIKV, 2 hours later, 10 mice were injected with 10 mg 6E7C4 mAb via tail vein, another 10 mice were injected with control rabbit IgG1 via tail vein. After virus injection, the mice were observed every 24 hours for their activity, and the body weight of mice was measured, when the body weight of mice decreased to or more than 25%, the mice were considered to be dead, and were executed by cervical dislocation after inhaled isoflurane ether anesthesia.
[0032] From Figure 3 It can be seen that 6E7C4 mAb can effectively protect mice against CHIKV East / Central / South African Ross strain infection, 10 mice did not show piloerection, weight loss, paralysis and other manifestations of disease, while 10 mice treated with control antibody showed rapid weight loss and all died.
[0033] The above in vitro and in vivo test results show that 6E7C4 rabbit anti-CHIKV E2 mAb can efficiently neutralize CHIKV infection and treat the mouse disease and death caused by CHIKV infection.
[0034] The above shows and describes the main features of the present application and the advantages of the present application. Those skilled in the art should understand that the present application is not limited by the above examples, the above examples and the description in the specification are only to illustrate the principles of the present application, and various changes and improvements can be made without departing from the spirit and scope of the present application, and these changes and improvements all fall within the scope of the claimed present application. The scope of protection of the present application is defined by the appended claims and their equivalents.
Claims
1. A monoclonal antibody against chikungunya virus, characterized in that: The amino acid sequence of the antibody includes an antibody heavy chain and an antibody light chain; wherein the amino acid sequence of the antibody heavy chain is shown in SEQ ID NO:2; and the amino acid sequence of the antibody light chain is shown in SEQ ID NO:
4.
2. The chikungunya virus monoclonal antibody according to claim 1, characterized in that: The antibody can block the infection of target cells by chikungunya virus in vitro.
3. The chikungunya virus monoclonal antibody according to claim 1, characterized in that: The antibody can protect against lethal infection by the chikungunya virus in vivo.
4. The chikungunya virus monoclonal antibody according to claim 1, characterized in that: Application in the development of therapeutic or preventative antibody drugs against Chikungunya virus infection.
5. A pharmaceutical composition, characterized in that, Contains the monoclonal antibody as described in claim 1.