Development and validation of relative activity evaluation method for SARS-CoV-2 neutralizing antibody based on Design of Experiment
10.13200/j.cnki.cjb.004319
- VernacularTitle:基于实验设计的SARS-CoV-2中和抗体相对活性检测方法的建立及验证
- Author:
LI Tiantian
- Publication Type:Journal Article
- Keywords:
Design of Experiment(DOE);
SARS-CoV-2;
Variant strain;
Pseudovirus;
Neutralizing antibody;
Relative activity
- From:
Chinese Journal of Biologicals
2024;37(10):1245-1250
- CountryChina
- Language:Chinese
-
Abstract:
Objective To develop and validate an evaluation method based on Design of Experiment(DOE) for the relative activity of SARS-CoV-2 neutralizing antibody, so as to provide a reliable detection method for the development and quality evaluation of SARS-CoV-2 neutralizing antibody.Methods A method for evaluating the relative activity of SARS-CoV-2neutralizing antibody was developed by using SARS-CoV-2 variant pseudovirus. The cell density [(3-5) × 10~5cells/mL],pseudovirus titer(3 750-6 250 TCID_(50)/mL), neutralization time of antibody to pseudovirus(0. 5-1. 5 h), and interaction time of antibody-pseudovirus complex and cells(48-96 h) were optimized by DOE. The optimized method was verified for the specificity, linear range, accuracy and precision, and used to detect the relative activity of two batches of SARS-CoV-2neutralizing antibodies.Results The optimum cell density was 4 × 10~5cells/mL, the titer of pseudovirus was 6 250 TCID_(50)/mL,the neutralization time of antibody to pseudovirus was 1. 5 h, and the interaction time of antibody-pseudovirus complex and cells was 96 h. Only SARS-CoV-2 neutralizing antibody showed a typical dose-response curve, while anti-VEGF monoclonal antibody and sample diluent did not show a dose-response curve. The theoretical titer of SARS-CoV-2 neutralizing antibody in the range of 50%-200% showed a good linear relationship with the detection value, and the linear equation was y = 1. 034 2 x-0. 013,r = 0. 958 8. The relative bias of the mean values of the three test results of the samples at the relative titer levels of 50%, 71%, 100%, 141% and 200% ranged from 6. 2% to 7. 3%, and the geometric coefficient of variation(GCV)ranged from 3. 2% to 16. 9%. The relative activity of the two batches of SARS-CoV-2 neutralizing antibodies was 91% and118% respectively.Conclusion The developed method has good specificity, accuracy and precision, and can be used for the detection of the relative activity of SARS-CoV-2 neutralizing antibody.
