CN114685628B - Antigen epitope peptide of RBD of SARS-CoV-2 and its application - Google Patents
Antigen epitope peptide of RBD of SARS-CoV-2 and its application Download PDFInfo
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Abstract
The invention provides an antigen epitope peptide of RBD of SARS-CoV-2, which comprises antigen dominant epitopes with amino acid sequences of SEQ ID NO. 11 and/or SEQ ID NO. 17. The invention also provides the application of the compound in preparing medicines for diagnosing, preventing or treating SARS-CoV-2 infection. The antigen potential epitope peptide provided by the invention has the effect of inhibiting the combination of RBD of SARS-CoV-2 and human angiotensin converting enzyme 2 (ACE 2), and can be used for preparing high-efficiency, low-toxicity and high-safety RBD-based medicaments for preventing SARS-CoV-2 infection.
Description
Technical Field
The invention relates to the technical field of biological medicine, in particular to the technical field of virus antigens, and especially relates to an epitope peptide of RBD of SARS-CoV-2 and application thereof.
Background
The novel coronavirus (Severe Acute Respiratory Syndrome Coronavirus, SARS-CoV-2, abbreviated as New crown) has serious infection hazard, and epidemic situation is still in global sustainable development, which brings serious challenges to epidemic situation prevention and control in China.
Since no specific new crown therapeutic drug exists at present, a vaccine is an ideal choice for controlling new crown infection. Although several new coronavirus vaccines are currently marketed, the constant emergence of new coronavirus variant strains such as Delta and Omicron strains, challenges the effectiveness and persistence of existing vaccination. The main initial part of the new crown infection is respiratory mucosa, and most of the existing new crown vaccines are immune by intramuscular injection (intramuscular injection for short), the immune response of the respiratory mucosa induced by the new crown vaccine is limited, and the intramuscular injection can not prevent the virus transmission of the host mucosa part in time. The choice of a suitable adjuvant and an immune pathway while inducing respiratory mucosa and systemic immune responses is a critical issue to be resolved by the new crown vaccine. Analyzing the effective components of SARS-CoV-2 antigen to induce protective response under different adjuvants and immune approaches and its immune response mechanism is the basis and premise for solving the scientific problem.
Spike protein (S protein or Spike protein) is an important structural protein of SARS-CoV-2, and is an antigen component of other vaccines except for inactivated vaccines in the existing novel crown vaccine. The Spike protein contains two subunits S1 and S2, wherein the C-terminal end of the S1 subunit contains a receptor binding domain (Receptor binding domain, abbreviated as RBD domain), and recognizes the receptor angiotensin converting enzyme E2 (ACE 2) on the surface of host cells via its RBD domain, mediating viral entry into host cells, and amino acid variation of the RBD domain leads to changes in the species specificity and infection characteristics of the virus. RBD contains the most immunodominant neutralizing epitope in the whole SARS-CoV-2 virus, and 90% of neutralizing antibodies in serum of the COVID-19 restorer are induced by RBD. Neutralizing antibodies targeting RBD have potential value in the prevention and treatment of SARS-CoV-2 infection. RBD can circumvent the ADE effect caused by NTD domain antibodies of the S1 subunit. In addition, RBD has stable property, easy recombinant expression and purification preparation, thus being an ideal antigen of the novel crown vaccine.
Studies have demonstrated that antibodies with neutralizing capacity play a protective role in the protection of new coronaviruses. The titer of neutralizing antibodies to SARS-CoV-2 is correlated with clinical outcome. Since protein antigens exert their function primarily by virtue of epitopes therein, the primary role played is "immunodominant" epitopes. Thus, the identification of protective epitopes for immunodominant responses in RBD antigens is an important premise for enhancing and optimizing the design of SARS-CoV-2 vaccines based on RBD antigens. Screening for immunodominant epitopes of RBD is a precondition for eliciting a more potent RBD immune response. Currently known B-cell epitopes of RBDs, either as deduced by bioinformatics software, or as identified by monoclonal antibodies, or as identified in a human or animal immune model, have not been reported for the comprehensive screening of RBDs involved in immune responses in different immune pathways. Since the primary site of infection of the new coronavirus is primarily the respiratory mucosa, there is a need to establish an accurate and effective method for screening and identifying B cell "dominant epitopes" of RBDs involved in immune responses under the "nasal drip route".
Disclosure of Invention
The present invention provides a method for identifying dominant epitope peptide of antibody of RBD of SARS-CoV-2 and application of said epitope peptide in preparing medicine for diagnosing, preventing and/or curing SARS-CoV-2 infection.
The invention firstly provides an antigen epitope peptide of RBD of SARS-CoV-2, which comprises antigen dominant epitopes with amino acid sequences of SEQ ID NO. 11 and/or SEQ ID NO. 17.
In one embodiment according to the invention, the epitope peptide is coupled to a polypeptide tag at the N-terminus or the C-terminus; preferably, the polypeptide label is a biotin label or a fluorescent label.
The present invention further provides the application of the antigen epitope peptide in preparing specific antibody diagnosis reagent for SARS-CoV-2 infection.
The present invention also provides the application of the antigen epitope peptide in preparing vaccine for preventing and treating SARS-CoV-2 infection.
The present invention further provides the application of the antigen epitope peptide as a screening target of SARS-CoV-2 infection resisting medicine.
In one embodiment according to the present invention, there is also provided a pharmaceutical composition for preventing or treating SARS-CoV-2 infection, comprising the above-described epitope peptide and a pharmaceutically acceptable adjuvant.
In one embodiment according to the invention, an adjuvant is also included, preferably adavax.
In one embodiment according to the invention, the pharmaceutical composition is in a dosage form for nasal administration; preferably in the form of spray, nasal drops, powder, gel or microsphere.
The invention further provides a diagnostic reagent for SARS-CoV-2 infection, which comprises the antigen epitope peptide, wherein the antigen epitope peptide is coated on a detection carrier, and the detection carrier is selected from any one of a polystyrene micro-reaction plate, a colloidal gold reagent strip, magnetic beads and a microfluidic chip.
In one embodiment according to the invention, a second antibody specifically recognizing a murine IgG antibody is further comprised;
preferably, the second antibody is selected from one of goat anti-mouse monoclonal antibody and rabbit anti-mouse polyclonal antibody;
preferably, the second antibody is conjugated with a ligand group that activates or quenches the specific fluorescent group.
The technical scheme of the invention has the following beneficial effects:
the RBD antibody immunodominant epitope peptide obtained by the method has the effect of inhibiting the combination of RBD and human angiotensin converting enzyme 2 (ACE 2), and can be used for preparing high-efficiency, low-toxicity and high-safety RBD-based medicaments for preventing SARS-CoV-2 infection, such as preventive vaccines.
The antibody dominant epitope peptide of RBD of SARS-CoV-2 provided by the invention has the effect of inhibiting the combination of RBD and human vascular tensein convertase 2 (ACE 2), animals are immunized by using the dominant epitope, immune preparation has no irrelevant components or harmful components, and the specific monoclonal antibody prepared by the antibody dominant epitope peptide can better prevent SARS-CoV-2 infection.
The antibody dominant epitope peptide of RBD of SARS-CoV-2 provided by the invention is sequence-conserved in various SARS-CoV-2 strains, so that it can be used as diagnostic reagent of SARS-CoV-2 or used for preventing and curing other SARS-CoV-2 infection.
Drawings
FIG. 1 is a graph showing the results of ELISA assays for overlapping peptides screened in accordance with the present invention using RBD nasal drip immunized BALB/c mice as primary antibodies;
FIG. 2 is a serum specific IgG antibody titer profile of BALB/c mice immunized with RBD nasal drops;
FIG. 3 is a serum-specific IgG antibody subtype profile of BALB/c mice immunized with RBD nasal drops;
FIG. 4 is a graph showing the results of neutralizing antibody titer analysis of the antiserum of RBD immunized BALB/c mice screened in accordance with the present invention for the inhibition of RBD binding to human vascular tensein convertase 2 (ACE 2);
FIG. 5 is a graph showing the results of a serum dilution reciprocal plot of the effect of antisera from RBD immunized BALB/c mice screened in accordance with the present invention on inhibiting RBD binding to human vascular tensein convertase 2 (ACE 2);
FIG. 6 shows the immunodominant epitope peptide RBD of the antibodies selected according to the present invention 61-78 、RBD 97-114 Analyzing a result map of the position distribution in the RBD three-dimensional structure;
FIG. 7 shows the immunodominant epitope peptide RBD of the antibodies selected according to the present invention 61-78 、RBD 97-114 Results of the amino acid sequence conservation analysis of (2).
Detailed Description
In order to make the technical problems, technical solutions and advantages to be solved more apparent, the following detailed description will be given with reference to the accompanying drawings and specific embodiments.
The experimental methods used in the following examples are conventional methods unless otherwise specified. Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.
Example 1 acquisition of overlapping peptides
Based on the RBD protein Sequence (Sequence ID: 6XDG_E), another desired overlapping peptide was obtained again by shifting downstream by an amino acid number smaller than the length of the overlapping peptide at a time relative to the length of the desired overlapping peptide. Wherein the length of the expected overlapping peptide can be 15-30 amino acids, the number of amino acids per step can be 4-8, in this example, starting from amino acid 1, 6 amino acids per step, and the overlapping 18 amino acid polypeptides (Shanghai Jier Biochemical technologies Co., ltd.) are synthesized to obtain 35 overlapping peptides. The purity is more than 95 percent. The synthetic walking overlapping peptide information is shown in table 1. The synthesized peptide is dissolved to a storage concentration of 1mg/mL by dimethyl sulfoxide (DMSO), and is frozen at-70 ℃ after subpackaging, and diluted to 1mM by PBS when in use.
TABLE 1 step overlapping peptide information
EXAMPLE 2 collection and preservation of RBD immune serum
Recombinant SEB whole protein of mutant SEB three toxin sites was constructed by the method disclosed based on the reference (reference: jintaozou et al a-Hemolysin-Aided Oligomerization of the Spike Protein RBD Resulted in Improved Immunogenicity and Neutralization Against SARS-CoV-2 Variants ariants.Front Immunol.2021 Sep 24;12:757691.) and the RBD protein dose of immunized mice was adjusted to 20 ug/dose, the dose of adjuvant AddaVax was 50 uL, the immunization route was nasal drip immunization, and three times over 0, 14, 21 days. Measuring the titer of the antiserum, and selecting RBD titer to be more than 1:64000 mice immune serum was used for subsequent detection.
EXAMPLE 3 screening of B cell immunodominant epitopes of RBD
The overlapping peptide coating concentration was adjusted to 5. Mu.g/well (using whole protein RBD (SSequence ID: 6XDG_E) as a positive control), the wells were coated, washed, blocked, and washed again, then RBD immune antiserum obtained in example 2 was added, the dilution was 1:50, incubation was performed for 1.5 h-washing, HRP-goat anti-mouse IgG (purchased from Enjing organism, cat# E1WP 319) was added, the dilution was 1:5000, TMB substrate chromogenic solution (purchased from Beyotime/Biyun, cat# P0209-100 ml) was added after washing, the OD value was read at 450nm after termination of the reaction, and the positive overlapping peptide was obtained by the formula 18 amino acid overlapping peptide OD detection value-blank control detection value)/(negative peptide OD detection value-blank control detection value). Gtoreq.2.1. Through SPSS16.0 data inspection, positive overlapping peptides with significant statistical significance relative to other positive overlapping peptide readings are obtained and are defined as immunodominant epitope peptides of B cells, namely immunodominant epitope peptides. Unrelated peptide OVA 192–201 (SEQ ID NO:36 EDTQAMPFRV) is an anionic control peptide.
Results: as shown in FIG. 1, there are 2 positive polypeptides RBD 61-78 (SEQ ID NO:11 CYGVSPTKLNDLCFTNVY)、RBD 97-114 (SEQ ID NO:17 TGKIADYNYKLPDDFTGC) has a statistically significant meaning relative to other dominant peptide reads, defined as dominant epitopes, OVA in this figure 192-201 A negative irrelevant peptide; by the method, not only all B cell epitopes of RBD are obtained by screening, but also B cell dominant epitopes of RBD are defined.
Example 4 antisera from RBD immunized mice were analyzed for their ability to inhibit RBD binding to human angiotensin converting enzyme 2 (ACE 2);
proteins from RBD were immunized by three intramuscular injections on days 0, 14, 21 with AddaVax adjuvant (purchased from InvivoGen) for 6-8 weeks BALB/c mice, day 7 post last immunization, and antisera were collected from the tail vein.
To evaluate the immunogenicity of RBD, mice were immunized by nasal drip with the RBD protein described in example 2 supplemented with adavax adjuvant, and post-trisection mice were collected from venous blood samples of the tail for RBD-specific antibody level detection. Results of RBD-specific IgG antibody titers from immunized mouse serum as shown in fig. 2, the geometric mean titers of RBD-specific IgG antibodies from immunized group serum increased to about 106. Wherein the log10 log of RBD specific IgG antibody titers was 5.95±0.2375 (P < 0.05). The above results indicate that: RBD can induce high levels of RBD specific antibodies by nasal drip immunization with adavax adjuvant.
The analysis results of the RBD specific IgG antibody subtype of the immunized mouse serum are shown in FIG. 3, wherein the RBD specific IgG antibody mainly comprises the IgG1 subtype.
Neutralizing antibody titers were first measured by competition ELISA methods to assess RBD-induced functional antibody levels. The competition inhibition assay was performed using a kit (Anti-SARS-CoV-2 Neutralizing Antibody Titer Serologic Assay Kit, available from Acrobiosystems, beijing, china). Serum samples from immunized mice were diluted 1:10 and mixed with RBD of 0.3 μg/ml HRP-SARS-CoV-2 spike protein for repeated analysis. The microplate reader detects the Optical Density (OD) at 450nm and calculates the neutralization inhibition according to the formula (1-average OD of sample/average OD of negative control) ×100%. Results are shown in FIG. 4, 20 μg of antibody NT50 (Titers of 50% Neutralization) induced after RBD 3 immunizations reached 1:3412 (P < 0.05), and the inverse serum dilution plot is shown in FIG. 5. The above results illustrate: RBD was boosted with adavax adjuvant and the induced high level of RBD-specific antibodies had the ability to compete for inhibition of RBD binding to human angiotensin converting enzyme 2 neutralization.
Example 5 position of immunodominant epitope peptide in three-dimensional Structure of RBD holoprotein
The present example is used to demonstrate the results of the positional distribution analysis of the immunodominant epitope peptide of antibodies in the three-dimensional structure of the RBD holoprotein.
Downloading a 3D structure diagram of the reported RBD protein in a pubMed protein public database, and marking the sequence positions of the immunodominant epitope peptides screened in the experiment by using PyMOL 1.1 program software.
The results are shown in FIG. 6, which shows the dominant peptide RBD 61-78 、RBD 97-114 Respectively located in different loop regions of the RBD three-dimensional crystal structure, it can be seen that the dominant epitope peptide sequence (antibody dominant epitope) is a reliable candidate molecule for RBD epitope vaccine.
EXAMPLE 6 amino acid sequence conservation analysis of immunodominant epitope peptide in each strain of SARS-CoV-2
This example is for antibody immunodominant epitope peptide RBD screened by the present invention 61-78 、RBD 97-114 Results of amino acid sequence conservation analysis of (2).
The amino acid sequence of RBD protein of each strain of 30 SARS-CoV-2 was searched in Genbank database, amino acid sequence alignment was performed using NCBI Basic Local Alignment Search Tool (BLAST) software, 30 strains of SARS-CoV-2 were randomly selected for multiple sequence alignment (Multiple Alignment), and the site address was https:// BLAST.
The results are shown in FIG. 7, which shows the dominant peptide RBD 61-78 、RBD 97-114 The amino acid sequences in each strain of 30 SARS-CoV-2 are conserved, so that it has good application prospect.
While the foregoing is directed to the preferred embodiments of the present invention, it will be appreciated by those skilled in the art that various modifications and adaptations can be made without departing from the principles of the present invention, and such modifications and adaptations are intended to be comprehended within the scope of the present invention.
Sequence listing
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<400> 17
Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr
1 5 10 15
Gly Cys
<210> 18
<211> 18
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 18
Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala Trp
1 5 10 15
Asn Ser
<210> 19
<211> 18
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 19
Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp
1 5 10 15
Ser Lys
<210> 20
<211> 18
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 20
Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn
1 5 10 15
Tyr Asn
<210> 21
<211> 18
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 21
Asn Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg
1 5 10 15
Leu Phe
<210> 22
<211> 18
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 22
Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn
1 5 10 15
Leu Lys
<210> 23
<211> 18
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 23
Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg
1 5 10 15
Asp Ile
<210> 24
<211> 18
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 24
Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile
1 5 10 15
Tyr Gln
<210> 25
<211> 18
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 25
Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr
1 5 10 15
Pro Cys
<210> 26
<211> 18
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 26
Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly Val Glu
1 5 10 15
Gly Phe
<210> 27
<211> 18
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 27
Ala Gly Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe
1 5 10 15
Pro Leu
<210> 28
<211> 18
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 28
Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly
1 5 10 15
Phe Gln
<210> 29
<211> 18
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 29
Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly
1 5 10 15
Val Gly
<210> 30
<211> 18
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 30
Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr
1 5 10 15
Arg Val
<210> 31
<211> 18
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 31
Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser
1 5 10 15
Phe Glu
<210> 32
<211> 18
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 32
Tyr Gln Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala
1 5 10 15
Pro Ala
<210> 33
<211> 18
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 33
Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly
1 5 10 15
Pro Lys
<210> 34
<211> 18
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 34
Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Thr Asn
1 5 10 15
Leu Val
<210> 35
<211> 19
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 35
Thr Val Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys
1 5 10 15
Val Asn Phe
<210> 36
<211> 10
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 36
Glu Asp Thr Gln Ala Met Pro Phe Arg Val
1 5 10
Claims (14)
1. An epitope peptide of RBD of SARS-CoV-2, characterized in that the amino acid sequence of the epitope peptide is SEQ ID NO. 11 or SEQ ID NO. 17.
2. The epitope peptide of claim 1, wherein said epitope peptide is conjugated to a polypeptide tag at the N-terminus or C-terminus.
3. The epitope peptide of claim 2, wherein said polypeptide tag is a biotin tag or a fluorescent tag.
4. Use of the epitope peptide of any one of claims 1-3 for the preparation of a diagnostic reagent for specific antibodies against SARS-CoV-2 infection.
5. Use of an epitope peptide according to any one of claims 1-3 in the manufacture of a vaccine for the prevention or treatment of SARS-CoV-2 infection.
6. A pharmaceutical composition for preventing or treating SARS-CoV-2 infection, comprising the epitope peptide of any one of claims 1-3 and a pharmaceutically acceptable adjuvant.
7. The pharmaceutical composition of claim 6, further comprising an adjuvant.
8. The pharmaceutical composition of claim 7, wherein the adjuvant is adavax.
9. The pharmaceutical composition according to any one of claims 6-8, wherein the pharmaceutical composition is in a dosage form for nasal administration.
10. The pharmaceutical composition of claim 9, wherein the dosage form is a spray, nasal drop, powder, gel formulation, or microsphere formulation.
11. A diagnostic reagent for SARS-CoV-2 infection, comprising the epitope peptide of any one of claims 1-3 coated on a detection carrier selected from any one of a polystyrene microplate, a colloidal gold reagent strip, magnetic beads and a microfluidic chip.
12. The diagnostic reagent of claim 11, further comprising a secondary antibody that specifically recognizes a murine IgG antibody.
13. The diagnostic reagent of claim 12, wherein the second antibody is selected from one of a goat anti-mouse monoclonal antibody and a rabbit anti-mouse polyclonal antibody.
14. The diagnostic reagent of claim 12, wherein the second antibody is conjugated with a ligand group that activates or quenches a specific fluorescent group.
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Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111830258A (en) * | 2020-07-03 | 2020-10-27 | 浙江睦科生物科技有限公司 | Novel coronavirus specific antibody detection kit |
| CN112646006A (en) * | 2021-01-20 | 2021-04-13 | 中国人民解放军陆军军医大学 | Marker epitope polypeptide for diagnosing COVID-19 mild and severe symptoms and application thereof |
| CN112851770A (en) * | 2021-02-05 | 2021-05-28 | 中国人民解放军陆军军医大学 | Alpha hemolysin epitope peptide for diagnosing or preventing staphylococcus aureus infection and application thereof |
| CN112940087A (en) * | 2021-03-17 | 2021-06-11 | 郑州大学 | Common epitope peptide of SARS-CoV and SARS-CoV-2 and its application |
| CN112961223A (en) * | 2021-02-24 | 2021-06-15 | 东南大学 | SARS-CoV-2 lymphocyte antigen epitope peptide and its application |
| KR20210123155A (en) * | 2020-04-02 | 2021-10-13 | 조선대학교산학협력단 | Use of RBD as diagnostic, treatment or vaccine of COVID-19 |
| WO2021209035A1 (en) * | 2020-04-17 | 2021-10-21 | 暨南大学 | Novel coronavirus t cell epitope peptide, and pmhc and preparation and application thereof |
| CN114058593A (en) * | 2020-08-06 | 2022-02-18 | 中国人民解放军陆军军医大学 | Monoclonal antibody of N antigen of SARS-CoV-2, detection method and use thereof |
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2022
- 2022-03-24 CN CN202210296306.0A patent/CN114685628B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20210123155A (en) * | 2020-04-02 | 2021-10-13 | 조선대학교산학협력단 | Use of RBD as diagnostic, treatment or vaccine of COVID-19 |
| WO2021209035A1 (en) * | 2020-04-17 | 2021-10-21 | 暨南大学 | Novel coronavirus t cell epitope peptide, and pmhc and preparation and application thereof |
| CN111830258A (en) * | 2020-07-03 | 2020-10-27 | 浙江睦科生物科技有限公司 | Novel coronavirus specific antibody detection kit |
| CN114058593A (en) * | 2020-08-06 | 2022-02-18 | 中国人民解放军陆军军医大学 | Monoclonal antibody of N antigen of SARS-CoV-2, detection method and use thereof |
| CN112646006A (en) * | 2021-01-20 | 2021-04-13 | 中国人民解放军陆军军医大学 | Marker epitope polypeptide for diagnosing COVID-19 mild and severe symptoms and application thereof |
| CN112851770A (en) * | 2021-02-05 | 2021-05-28 | 中国人民解放军陆军军医大学 | Alpha hemolysin epitope peptide for diagnosing or preventing staphylococcus aureus infection and application thereof |
| CN112961223A (en) * | 2021-02-24 | 2021-06-15 | 东南大学 | SARS-CoV-2 lymphocyte antigen epitope peptide and its application |
| CN112940087A (en) * | 2021-03-17 | 2021-06-11 | 郑州大学 | Common epitope peptide of SARS-CoV and SARS-CoV-2 and its application |
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| CN114685628A (en) | 2022-07-01 |
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