Neutralizing antibody kit for detecting epitope peptide antigen screened based on binding structural domain of novel coronavirus RBD and ACE2 receptor
An epitope peptide and kit technology, applied in the field of viruses, can solve the problems of inability to distinguish between total binding antibodies and neutralizing antibodies, cumbersome and time-consuming, 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 domain
[0068] 1. Experimental method
[0069] 64 epitope peptide antigens were screened by computer simulation of the RBD and ACE2 receptor binding domains of the new coronavirus, and the screened epitope peptides were chemically synthesized and coated on microwell plates, and 8 effective epitopes were screened out by ELISA experiments Peptide antigen, other epitope peptide ELISA experiments were negative.
[0070] 2. Experimental results
[0071] Among them, the verification results of 4 epitope peptide antigens are consistent with the test results of the specific neutralizing antibody competition 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] E3: GGNYNYLYRLFRKSNLKPF...
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 the screening of the new coronavirus RBD and ACE2 receptor binding domain are as follows:
[0079] 1. A solid phase carrier coated with epitope peptide antigens (amino acid sequences such as epitope peptides E1, E2, E3, and E4 shown in SEQ ID NO: 1-4) screened by the new coronavirus RBD and ACE2 receptor binding domain 96-well plate / 48-well plate, each epitope peptide antigen is 400ng / well (preparation method: epitope peptide antigen (E1, E2, E3, E4) is coated in a microwell plate, and each well contains 4 Each 400ng of epitope peptides was coated on an enzyme-linked plate, coated overnight at 4°C, and blocked with 3% skimmed milk powder for 2h),
[0080] Wherein, E1: PGQTGKIADYNYKLPDDFTGGGG-Lys-Biotin (the amino acid sequence is shown in SEQ ID NO: 1);
[0081] E2: Biotin-Ahx-G...
Embodiment 3
[0099] The detection of embodiment 3 clinical sample
[0100] 1. Experimental method
[0101] 1. In order to test the detection effect of the kit of Example 1 in vaccinators, the kit of Example 1 was used to detect clinical samples. In this example, 5 vaccinators (all of whom were confirmed to have neutralizing antibodies in their serum by neutralizing antibody competition experiments) were included, and 8 healthy individuals were also included as controls at the same time.
[0102] 2. Test results
[0103] The test results are shown in Table 1. The S / N values of the healthy controls were all less than 1.2, while the S / N values of the vaccinators were all greater than 2, that is, the kit can detect the neutralizing antibodies present in the sera of the vaccinators
[0104] Table 1:
[0105]
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