Development and verification of a fluorescence quantitative detection method for SARS-CoV-2 antibodies based on magnetic bead sorting and peptide self-assembly
10.13200/j.cnki.cjb.003909
- VernacularTitle:基于磁珠分选及多肽自组装介导的SARS-CoV-2抗体荧光定量检测方法的建立及验证
- Author:
JIN Yu
- Publication Type:Journal Article
- Keywords:
Severe acute respiratory syndrome coronavirus 2(SARS-CoV-2);
Antibody;
Magnetic bead sorting;
Self-assembly of peptides
- From:
Chinese Journal of Biologicals
2024;37(10):1257-1262
- CountryChina
- Language:Chinese
-
Abstract:
Objective To develop and verify a fluorescence quantitative detection method for antibody against severe acute respiratory syndrome coronavirus 2(SARS-CoV-2) based on magnetic bead sorting and peptide self-assembly. Methods Magnetic beads were specifically bound to nucleocapsid(N) protein of SARS-CoV-2 by click chemistry reaction, which were detected by Western blot and BCA. Using magnetic beads labeled with N protein as coating antigen and donkey anti-human IgG labeled with alkaline phosphatase(ALP) as enzyme-labeled antibody,combined with assemblable peptide and thioflavine-T(ThT)luminescence system, a quantitative detection method for SARS-CoV-2 antibody was established, and the specificity,linear range, limit of detection(LOD),precision and durability were verified. The developed method and commercial ELISA kit were used to detect 8 serum samples with known background respectively,and the consistency between them was calculated.Results Most of SARS-CoV-2 N proteins were stably bound to magnetic beads. The developed method specifically detected positive serum samples. The concentration of SARS-CoV-2 IgG antibody standard ranged from 2. 5 to 80 ng/mL,showing a good linear relationship with fluorescence value, and the linear equation was y = 182. 11 x + 5 095. 5, R~2= 0. 99;The LOD was 2. 5 ng/mL;The coefficient of variation(CV)within batches ranged from 3. 9% to 10. 6%,the CV between batches ranged from 6. 0% to 13. 8%, and the CV of durability was within 1. 3%～6. 2%. The consistency between the results of the developed method and commercial ELISA kit was high(R~2= 0. 99,P < 0. 000 1). Conclusion The developed fluorescence quantitative detection method based on magnetic bead sorting and peptide self-assembly showed good specificity,precision and durability, which might be used for quantitative detection of SARS-CoV-2 antibody.
