Neutralizing Antibody Kit for Epitope Peptide Antigen Detection Based on Screening of Novel Coronavirus RBD and ACE2 Receptor Binding Domain
A technology of epitope peptides and kits, applied in the field of viruses, can solve the problems of inability to distinguish total binding antibodies and neutralizing antibodies, cumbersome and time-consuming, etc., and achieve the effect of improving the detection rate of new crowns, less interference factors, and strong specificity
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Embodiment 1
[0067] Example 1 Epitope peptides screened by coronavirus RBD and ACE2 receptor binding domains
[0068] 1. Experimental method
[0069] 64 epitope peptide antigens were screened by computer simulation of the new coronavirus RBD and ACE2 receptor binding domains, and the screened epitope peptides were chemically synthesized and coated on the microplate respectively. After ELISA experiments, 8 effective epitopes were verified and screened. Peptide antigen, other epitope peptide ELISA experiments were negative.
[0070] 2. Experimental results
[0071] Among them, the results of 4 epitope peptide antigen verifications are consistent with the test results of the specific neutralizing antibody competition method ELISA kit for the SARS-CoV-2 RBD domain. The 4 epitope peptide antigens include:
[0072] E1: PGQTGKIADYNYKLPDDFTGGGG (the amino acid sequence is shown in SEQ ID NO: 1);
[0073] E2: GGGCVIAWNSNNLDSKVGGNYNY (the amino acid sequence is shown in SEQ ID NO: 2);
[0074] E...
Embodiment 2
[0076] Example 2 A new coronavirus neutralizing antibody detection kit
[0077] 1. Composition
[0078] The components of the Neutralizing Antibody Kit for Epitope Peptide Antigen Detection Based on New Coronavirus RBD and ACE2 Receptor Binding Domain Screening are as follows:
[0079] 1. A solid-phase carrier coated with epitope peptide antigens (epitope peptides E1, E2, E3, E4 whose amino acid sequences are shown in SEQ ID NOs: 1 to 4) screened by the RBD and ACE2 receptor binding domains of the novel coronavirus 96-well plate / 48-well plate, each epitope peptide antigen is 400ng / well (preparation method: epitope peptide antigens (E1, E2, E3, E4) are coated in the microwell plate, each well contains 4 400ng each of epitope peptides, coated on enzyme-linked plate, coated overnight at 4°C, and blocked with 3% nonfat milk powder for 2h),
[0080] Wherein, E1: PGQTGKIADYNYKLPDDFTGGGG-Lys-Biotin (amino acid sequence shown in SEQ ID NO: 1);
[0081] E2: Biotin-Ahx-GGGCVIAWNSNNLD...
Embodiment 3
[0099] Example 3 Detection of clinical samples
[0100] 1. Experimental method
[0101] 1. In order to test the detection effect of the kit of Example 1 in vaccine recipients, the kit of Example 1 was used to detect clinical samples. In this example, 5 vaccinated persons (all of which were confirmed to have neutralizing antibodies in serum by neutralizing antibody competition experiments) were included, and 8 healthy persons were also included as controls.
[0102] 2. Test results
[0103] The test results are shown in Table 1. The S / N values of the healthy controls are all less than 1.2, while the S / N values of the vaccine recipients are all greater than 2, that is, the kit can detect the neutralizing antibodies present in the serum of the vaccine recipients.
[0104] Table 1:
[0105]
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