Chikungunya virus envelope E2 protein monoclonal antibody and application thereof
By preparing a rabbit monoclonal antibody targeting the CHIKV E2 protein, the problem of the lack of highly effective Chikungunya virus treatment drugs in the existing technology has been solved, and the effects of blocking viral infection in vitro and protecting mice in vivo have been achieved.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2026-01-07
- Publication Date
- 2026-03-13
AI Technical Summary
There is a lack of effective drugs against chikungunya virus in the current technology, especially highly effective and broad-spectrum monoclonal antibodies, and there are few reports on the efficacy evaluation of monoclonal antibodies in non-human primates.
Rabbit monoclonal antibodies against the CHIKV E2 protein were prepared and developed as preventive or therapeutic antibodies against CHIKV infection by efficiently neutralizing the major prevalent CHIKV genotypes worldwide. The specific steps included immunizing New Zealand rabbits, sorting spleen cells, constructing expression plasmids, purifying the antibodies, and verifying their neutralizing activity.
It demonstrated highly efficient neutralizing activity and protective effect by blocking CHIKV infection of target cells in vitro and protecting mice against lethal infection in vivo.
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Abstract
Description
Technical Field
[0001] This invention relates to the field of biomedical engineering technology. Specifically, this invention provides an amino acid sequence of a monoclonal antibody against the E2 protein of the chikungunya virus envelope and its application in the development of therapeutic or preventive antibodies against chikungunya virus infection. Background Technology
[0002] Chikungunya virus (CHIKV) is the pathogen that causes Chikungunya fever. It belongs to the family Togaviridae, genus Alphavirus, and its genome is a single-stranded positive-sense RNA. CHIKV is an important arbovirus, primarily transmitted by Aedes mosquitoes. The general population is susceptible, and in addition to causing fever and rash, it often leads to chronic arthritis with a course lasting more than six months. In recent years, there have been reports of CHIKV causing more serious illnesses, including encephalitis and hemorrhagic diseases. Currently, CHIKV has spread to more than 100 countries and regions worldwide, putting 39% of the global population at risk of infection and causing millions of infections annually, representing a significant burden on global public health. There is currently no effective treatment for CHIKV infection.
[0003] The CHIKV genome encodes two open reading frames (ORFs). The 5' ORF encodes non-structural proteins (nsp1, nsp2, nsp3, nsp4), which are mainly responsible for the replication of genomic RNA; the 3' ORF encodes structural proteins (capsid protein C, envelope protein E3, envelope protein E2, envelope protein E1), which are responsible for the formation of viral particles.
[0004] The translation product of the structural protein is a polyprotein precursor. The capsid protein at the amino terminus of the polyprotein is released from the polyprotein through its own protease hydrolysis function, while the carboxyl-terminal polypeptide is cleaved by the host signal peptidase in the endoplasmic reticulum to form the precursor pE2 protein, 6K protein and E1 protein. Subsequently, pE2 is cleaved by furin protease in the host cell 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 residues, which interact with the negatively charged genomic RNA. The envelope protein E2 is the viral receptor-binding protein, primarily responsible for the binding of CHIKV to cell surface receptors. Structurally, the E2 protein is divided into four parts: an extracellular region, a stem region, a transmembrane region, and an intramembrane region. The CHIKV E2 protein consists of 423 amino acid residues (aa) in total, with the extracellular region approximately 260 aa, the stem region approximately 100 aa, the transmembrane region approximately 30 aa, and the intramembrane region approximately 33 aa. The extramembrane region plays a crucial role in viral adsorption to host cells and transmembrane infection, containing epitopes for neutralizing antibodies; the transmembrane region enters the lipid bilayer, and the intramembrane region is located inside the viral particle, interacting with the nucleocapsid protein. The E1 protein belongs to the second class of fusion proteins and is primarily responsible for membrane fusion. Structurally, E1 is also divided into four parts: an extramembrane region, a stem region, a transmembrane region, and an intramembrane region. E2 proteins form heterodimers that anchor to the envelope surface of viral particles.
[0006] CHIKV infection of cells begins with the binding of the virus to cell surface receptors. After the CHIKV E2 protein binds to the receptor, the viral particles form endosomes through receptor-mediated endocytosis and enter the cell. As the endosomes mature, acidification causes a conformational change in the CHIKV E2-E1 heterodimer, leading to the exposure of the fusion peptide in the E1 protein and its insertion into the endosome membrane, inducing the fusion of the viral envelope and the endosome membrane. After membrane fusion, 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, showing good results in treating Ebola virus, respiratory syncytial virus, and other viral infections. Monoclonal antibodies have emerged as a new class of highly effective drugs for treating viral infections and are currently a hot topic in antiviral drug development. Specific monoclonal antibodies against Chikungunya virus have attracted widespread attention as a potential novel treatment. Although some monoclonal antibodies against Chikungunya virus have been reported, their neutralizing activity is not strong enough, and there are very few reports on the efficacy of monoclonal antibodies evaluated in non-human primates. Therefore, developing new antibodies for the prevention and treatment of Chikungunya virus is of urgent practical significance in filling the gap in specific drugs for Chikungunya virus.
[0008] Compared to mouse monoclonal antibodies, rabbit monoclonal antibodies have a series of unique advantages in terms of molecular structure, immune mechanism, and engineering: 1. Broader immune recognition spectrum: Rabbits have a larger B cell library, enabling the generation of a wider and more diverse antibody spectrum, which is beneficial for recognizing more types of antigenic epitopes; 2. Higher affinity and specificity: Rabbit antibodies mature for affinity through a dual mechanism of high-frequency somatic hypermutation and unique gene conversion, and their complementarity-determining region structure is unique. These characteristics together make them generally exhibit higher affinity and specificity for antigens; 3. Stronger structural stability: Rabbit antibodies contain an additional disulfide bond in the variable region of the heavy chain, and there is also a linking disulfide bond between the variable region and the constant region. This unique disulfide bond network significantly enhances the structural stability of the molecule; 4. Easier humanization: Rabbit monoclonal antibodies are easy to humanize, and several humanized rabbit monoclonal antibody drugs have already been marketed. Summary of the Invention
[0009] CHIKV E2 protein is a key protein mediating viral invasion of host cells and is the main protein for inducing neutralizing antibodies. This invention uses CHIKV E2 protein as the target antigen to prepare rabbit monoclonal antibodies, obtaining a neutralizing antibody that can efficiently and broadly neutralize the major circulating genotypes of CHIKV worldwide, which can be further developed into a preventive or therapeutic antibody against CHIKV infection.
[0010] Specifically, this invention provides the amino acid sequence of a rabbit monoclonal antibody against CHIKV E2 protein and its uses. This monoclonal antibody can block CHIKV infection of target cells in vitro. In vivo, it can protect mice against lethal CHIKV infection. The deoxynucleotide sequences of the antibody heavy chain signal peptide and variable region gene are shown in SEQ ID NO:1, and the amino acid sequences of the antibody heavy chain signal peptide and variable region protein are shown in SEQ ID NO:2. The deoxynucleotide sequences of the antibody light chain signal peptide and variable region gene are shown in SEQ ID NO:3, and the amino acid sequences of the antibody light chain signal peptide and variable region protein are shown in SEQ ID NO:4.
[0011] More specifically, the present invention provides a monoclonal antibody against CHIKV envelope protein E2, characterized in that: the amino acid sequence of the antibody includes a variable region of the antibody heavy chain and a variable region of the antibody light chain; The amino acid sequence of the variable region of the antibody heavy chain is shown in SEQ ID NO:2; The amino acid sequence of the variable region of the antibody light chain is shown in SEQ ID NO:4.
[0012] Furthermore, the present invention provides a monoclonal antibody against the CHIKV envelope protein E2, characterized in that: The deoxynucleotide sequence of the variable region gene of the antibody heavy chain is shown in SEQ ID NO:1.
[0013] Furthermore, the present invention provides a monoclonal antibody against the CHIKV envelope protein E2, characterized in that: The deoxynucleotide sequence of the variable region gene of the antibody light chain is shown in SEQ ID NO:3.
[0014] Furthermore, the present invention provides a monoclonal antibody against the CHIKV envelope protein E2, characterized in that: The aforementioned antibody is used as a therapeutic or preventative ingredient, a drug, or for the preparation of a drug; The aforementioned therapeutic components / drugs can block CHIKV infection of target cells in vitro.
[0015] Furthermore, the rabbit monoclonal antibody against CHIKV envelope protein E2 provided by the present invention is characterized in that: The aforementioned antibody is used as a therapeutic or preventative ingredient, a drug, or for the preparation of a drug; The aforementioned therapeutic ingredients / drugs can protect mice against lethal CHIKV infection in vivo.
[0016] In short, this antibody can block CHIKV infection of target cells in vitro.
[0017] This antibody can protect mice from lethal infection with CHIKV in vivo.
[0018] Furthermore, the rabbit monoclonal antibody against CHIKV envelope protein E2 provided by the present invention is characterized in that: Application in the development of therapeutic or preventative antibodies against CHIKV infection. Attached Figure Description
[0019] Figure 1 ELISA detection of the binding of monoclonal antibody 4G5D6 to chikungunya virus envelope protein E2. Figure 2 Monoclonal antibody 4G5D6 neutralizes infection of Chikungunya virus strain 37997. Figure 3 Monoclonal antibody 4G5D6 protects mice against challenge with Chikungunya virus Ross strain (weight change graph). Figure 4 Monoclonal antibody 4G5D6 protects mice against challenge with Chikungunya virus Ross strain (survival rate graph). Detailed Implementation
[0020] The implementation of the present invention will be described in detail below with reference to the embodiments and accompanying drawings. The following embodiments are implemented based on the technical solution of the present invention, and specific implementation methods and operating procedures are given. However, the scope of protection of the present invention is not limited to the following embodiments. All operations involving CHIKV in the present invention are performed in the Biosafety Level 3 Laboratory of the Naval Medical University of the Chinese People's Liberation Army.
[0021] I. Main Experimental Materials 1. CHIKV E2 protein, recombinantly expressed in human embryonic kidney 293T cells, is derived from the East / Central / South Africa type Indian Ocean lineage LR2006 strain. This protein was recombinantly expressed in freestyle 293F cells grown in serum-free medium by the Department of Biomedical Protection, Naval Medical University of the Chinese People's Liberation Army.
[0022] 2. The CHIKV virus, including the Asian type 37997 strain and the East / Central / South Africa type Ross strain, was synthesized by the Department of Biomedical Protection, Naval Medical University of the Chinese People's Liberation Army using reverse genetics technology.
[0023] 3. C57BL / 6 mice were purchased from Shanghai Xipubikai Experimental Animal Co., Ltd.
[0024] 4. The mouse anti-CHIKV polyclonal antibody was prepared by immunizing mice with Ross strain CHIKV inactivated by β-propiolactone using the Department of Biomedical Protection, Naval Medical University of the Chinese People's Liberation Army.
[0025] II. Experimental Methods and Results 1. Preparation and identification process of rabbit monoclonal antibodies New Zealand rabbits were immunized with recombinant CHIKV E2 protein combined 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 thoroughly emulsified with 0.5 ml of complete Freund's adjuvant. The mixture was then administered subcutaneously at multiple sites. Booster immunizations were given at 3, 6, and 9 weeks after the initial immunization, for a total of three times. For booster immunizations, 0.1 mg of CHIKV E2 protein was used, with incomplete Freund's adjuvant administered subcutaneously. Three weeks after each immunization, blood was collected from the marginal ear vein, and IgG antibodies in the rabbit serum were detected using ELISA (method described below). After week 12, rabbits were anesthetized with isoflurane, and blood was collected via cardiac puncture. After the rabbits died, their spleens were harvested, cut into small pieces, ground and filtered on a nylon mesh to prepare a single-cell suspension. Single spleen cells expressing CHIKV E2 protein IgG antibodies were sorted using flow cytometry. These single spleen cells were seeded into 96-well plates and cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum, 100 U / ml penicillin, and 100 mg / ml streptomycin. The 96-well plates were placed in a 37°C cell culture incubator with 5% CO2 and saturated humidity for approximately 7-10 days. When the cells covered the bottom of the wells, the presence of CHIKV E2 IgG antibodies in the cell culture supernatant was detected by ELISA. Total RNA was extracted from antibody-positive cells, and the antibody heavy chain signal peptide and variable region gene (Signal-VH), and light chain signal peptide and variable region gene (Signal-VL) were amplified using reverse transcription polymerase chain reaction. PCR products were recovered by agarose gel electrophoresis and inserted into the pMD18T vector (Takara). After DNA sequencing, the antibody heavy chain Signal-VH was spliced with rabbit IgG1 CH1-CH2-CH3 to form a complete heavy chain gene, and the light chain Signal-VL gene was spliced with the rabbit light chain constant region gene to form a complete light chain gene. The full-length heavy chain and light chain genes were then inserted into the cloning site 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 1:2 mass ratio. The presence of CHIKV E2 protein IgG antibodies in the cell culture supernatant was detected by ELISA. The supernatant with positive IgG antibodies was used to determine its neutralizing activity using a viral microneutralization assay. If the monoclonal antibody showed viral neutralizing activity, recombinant expression was performed on freestyle 293F cells cultured in serum-free medium. Freestyle 293F cells were cultured to 300 ml (1L culture flask) to achieve a cell density of 1×10⁻⁶ cells / mL. 6At a concentration of / ml, the antibody heavy chain and light chain expression plasmids were co-transfected into freestyle 293F cells using polyethylenimine (PEI) reagent. After culturing the cells for 72 hours, the antibody was purified from the culture supernatant using Staphylococcus aureus Protein A affinity chromatography and subjected to ELISA, virus neutralization assay, and mouse challenge protection assay.
[0026] 2. ELISA detection of CHIKV E2 protein IgG antibody CHIKV E2 protein was diluted with 50 mM carbonate coating buffer (pH 9.6) to a concentration of 0.5 μg / ml. 0.1 ml of the solution was added to each well of an ELISA plate, containing 0.05 mg of CHIKV E2 protein. The plate was incubated overnight at 4°C. The next day, the coating buffer was removed, and each well was washed once with 0.2 ml of PBS buffer. Then, 0.2 ml of PBS buffer (blocking buffer, pH 7.4) containing 3% bovine serum albumin, 5% goat serum, and 0.05% Tween 20 was added to each well. After incubation at room temperature for 2 hours, the blocking buffer was removed, and the wells were washed three times with PBS buffer (washing buffer) containing 0.05% Tween 20. Finally, 0.1 ml of rabbit spleen cell culture supernatant diluted with blocking buffer, culture supernatant co-transfected with antibody expression plasmid into 293T cells, or monoclonal antibody purified from culture supernatant co-transfected with antibody expression plasmid into freestyle 293F cells was added. Incubate the wells at room temperature on a gently shaking horizontal shaker for 30 minutes. Then discard the reaction solution in the wells, wash the wells 5 times with washing buffer, and add horseradish peroxidase-labeled anti-rabbit IgG (H+L) (Thermo Biotech product) diluted 1:2000 with blocking buffer. Incubate the wells at room temperature on a gently shaking horizontal shaker for 30 minutes. Then discard the enzyme-labeled antibody dilution in the wells, wash the wells 5 times with washing buffer, and add 0.1 ml of substrate solution containing 5,5'-tetramethylbenzidine (TMB) to each well. Incubate the wells at room temperature in the dark for 5 minutes, then add 50 ml of 2 M sulfuric acid, mix well, and measure the absorbance at 450 nm using an ELISA reader. The reference wavelength is 630 nm.
[0027] Figure 1The ELISA absorbance values (A450-630) of different concentrations of a CHIKV E2 protein monoclonal antibody (numbered 4G5D6) identified through screening are shown. Rabbit IgG1 (Cell Signaling) was used as a negative control. The results indicate that monoclonal antibody 4G5D6 specifically binds to the CHIKV E2 protein. The deoxynucleotide sequence of the heavy chain variable region gene of this 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 deoxynucleotide 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 assays and in vivo mouse challenge protection assays were performed on this monoclonal antibody to evaluate its antiviral efficacy.
[0028] Specifically: SEQ ID NO:1: Deoxynucleotide sequence of the signal peptide and variable region of the monoclonal antibody 4G5D6 heavy chain (1-60 are the signal peptide deoxynucleotide sequence, 61-411 are the variable region deoxynucleotide sequence). 1 ATGGAGACTG GGCTGCGCTG GCTTCTCCTG GTCGCTGTGC TCAAAGGTGT CCAGTGTCAG 61 TCGGTGGAGG AGTCCGGGGG TCGCCTGGTC ACGCCTGGAG GATCCCTGAC ACTCACCTGC 121 ACAGTCTCTG GAATCGACCT CAGCAGCTAT GCAATGAGCT GGGTCCGCCA GGCTCCAGGG 181 AAGGGGCTGG AATGGATCGG AATCATTTAT AGGGATGGTA GCGCATCCTA CGCGAGCTGG 241 GCGAAAGGCC GATTCACCAT CTCCAAAACC TCGACCACGG TGGATCTGAA AAGCACCAGT 301 CCGACAACCG AGGACACGGC CACCTATTTC TGTGCCAGAG GTTATGCTGG TGGTGATGAT 361 TATGCTTATG TTAACATCTG GGGCCCAGGC ACCCTGGTCA CCGTCTCCTC A SEQ ID NO:2: Amino acid sequence of monoclonal antibody 4G5D6 heavy chain signal peptide and variable region (1-20 are amino acid sequences of signal peptide, 21-137 are amino acid sequences of variable region). 1 METGLRWLLL VAVLKGVQCQ SVEESGGRLV TPGGSLTLTC TVSGIDLSSY AMSWVRQAPG 61 KGLEWIGIIY RDGSASYASW AKGRFTISKT STTVDLKSTS PTTEDTATYF CARGYAGGDD 121 YAYVNIWGPG TLVTVSS SEQ ID NO:3: Deoxynucleotide sequence of the light chain signal peptide and variable region of monoclonal antibody 4G5D6 (1-69 are the deoxynucleotide sequence of the signal peptide, 70-399 are the deoxynucleotide sequence of the variable region). 1 ATGGACACGA GGGCCCCCAC TCAGCTGCTG GGGCTCCTGC TGCTCTGGCT CCCAGGTGCC 61 AGATGTGCTG ACATCGAGAT GACCCAGACT CCAGCCTCCG TGTCTGGAGC TGTGGGAGGC 121 ACAGTCACCA TCAAGTGCCA GGCCAGTCAG AACATTTACA GGAATTTAGC CTGGTATCAG 181 CAGAAACCAG GGCAGCGTCC CAAACTCCTG ATCTATGGTG CATTCAATTT AGAATCTGGG 241 GTCCCATCGC GGTTCAAAGG CAGTGGATCT GGGACAGAGT ACACTCTCAC CATCAGCGAC 301 CTGGAGTGTG ACGATGCTGC CACTTATTAT TGTCAACACT ATTATGGTCA TAATGTTTTT 361 GGTGCAGCTT TCGGCGGAGG GACCGAGGTG GTGGTCAAG SEQ ID NO:4: Amino acid sequence of the light chain signal peptide and variable region of monoclonal antibody 4G5D6 (1-23 are amino acid sequences of the signal peptide, 24-133 are amino acid sequences of the variable region). 1 MDTRAPTQLL GLLLLWLPGA RCADIEMTQT PASVSGAVGG TVTIKCQASQ NIYRNLAWYQ 61 QKPGQRPKLL IYGAFNLESG VPSRFKGSGS GTEYTLTISD LECDDAATYY CQHYYGHNVF 121 GAAFGGGTEV VVK 3. Neutralizing activity of monoclonal antibodies against CHIKV The cultured young hamster kidney (BHK) cells were passaged and seeded into 96-well plates. The culture medium was DMEM medium supplemented with 10% fetal bovine serum, 100 U / ml penicillin, and 100 mg / ml streptomycin (hereinafter referred to as complete DMEM medium). 10,000 cells were placed in each well and cultured in a cell culture incubator at 37°C with 5% CO2 and saturated humidity for 12 hours.
[0029] Rabbit monoclonal antibody 4G5D6 was serially diluted in 96-well plates (using complete DMEM medium as the diluent). Rabbit IgG1 (Cell Signaling) was used as a negative control. 50 ml of each concentration gradient was aspirated into each well of the 96-well plate. 50 ml of diluted CHIKV virus solution (containing approximately 500 focus-forming units (PFUs) of CHIKV, also using complete DMEM medium) was added to each well. The mixture was then placed in a 37°C cell culture incubator. After 30 minutes, the liquid was transferred to BHK cell culture wells (after removing the original cells and preparing the culture medium). The cells were cultured for another 18 hours, and the expression of CHIKV protein in the cells was detected by immunofluorescence. The specific procedure is as follows: Aspirate the culture medium from the culture plate, add 0.1 ml of methanol to each well, place the culture plate at -20°C for 20 minutes, remove the plate, aspirate the methanol, wash each well once with phosphate-buffered saline (PBS), then add 0.1 ml of PBS containing 3% bovine serum albumin (BSA) (hereinafter referred to as 3% BSA-PBS), place on a horizontal shaker, and shake slowly at room temperature for 1 hour. Aspirate the 3% BSA-PBS from the culture plate, add 0.1 ml of 1% BSA-PBS containing anti-CHIKV polyclonal antibody (antibody diluted 500 times) to each well, and shake slowly at room temperature for 1 hour. Aspirate the CHIKV polyclonal antibody working solution from the culture plate, wash each well 3 times with PBS, then add 100 ml of 1% BSA-PBS containing Alexa Fluor 488-labeled anti-mouse IgG (luciferin antibody diluted 1500 times), and shake slowly at room temperature in the dark for 1 hour. Aspirate the luciferin antibody working solution from the culture plate, and add 0.1 ml of DAPI nuclear staining solution to each well. ml, shake slowly at room temperature in the dark for 10 minutes, remove DAPI cell nuclear staining solution from the culture plate, wash each well 3 times with PBS, count the green fluorescent positive cells in each well using a cell imaging and analysis system (BioTek Cytation 5 Imaging Reader), and then calculate the neutralization percentage (%) = number of positive cells in the monoclonal antibody-treated well or control rabbit IgG-treated well / number of positive cells in the well without antibody.
[0030] from Figure 2 It is evident that the 4G5D6 monoclonal antibody can effectively neutralize the infection of target cells BHK by 37,997 Asian-type CHIKV strains.
[0031] 4. Evaluate the in vivo antiviral activity of the monoclonal antibody using a mouse virus challenge model. Twenty 8-week-old female C57BL / 6 mice were randomly divided into two groups of 10 each. Each group was intranasally infected with the CHIKV East / Central / South Africa Ross strain, with a volume of 50 ml containing 1×10⁻⁶ CHIKV. 6Two hours after administering CHIKV to PFUs, 10 mice were injected intraperitoneally with 10 mg of 4G5D6 monoclonal antibody via the tail vein, while another 10 mice were injected intravenously with control rabbit IgG1. Following virus injection, the mice's activity level and body weight were monitored every 24 hours. Mice were considered dead when their body weight decreased by 25% or more, and were euthanized by inhaled isoflurane anesthesia and cervical dislocation.
[0032] from Figure 3 It is evident that the 4G5D6 monoclonal antibody can effectively protect mice against infection with the CHIKV East / Central / South Africa Ross strain. None of the 10 mice showed symptoms such as piloerection, weight loss, or paralysis, while all 10 mice treated with the control antibody rapidly lost weight and died.
[0033] The above in vitro and in vivo test results all indicate that the 4G5D6 rabbit anti-CHIKV E2 monoclonal antibody can effectively neutralize CHIKV infection and treat CHIKV-induced morbidity and mortality in mice.
[0034] The foregoing has shown and described the main features and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited to the above embodiments. The embodiments and descriptions in the specification are merely illustrative of the principles of the invention. Various changes and modifications can be made to the invention without departing from its spirit and scope, and all such changes and modifications fall within the scope of the present invention as claimed. The scope of protection of the present invention is defined by the appended claims and their equivalents.
Claims
1. A monoclonal antibody against the E2 protein of the chikungunya virus envelope, 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 monoclonal antibody against the E2 protein of the chikungunya virus envelope as described in claim 1, characterized in that: The antibody can block the infection of target cells by chikungunya virus in vitro.
3. The monoclonal antibody against the E2 protein of the chikungunya virus envelope as described in claim 1, characterized in that: The antibody can protect against lethal infection by the chikungunya virus in vivo.
4. The monoclonal antibody against the E2 protein of the chikungunya virus envelope as described in 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.