Verification of a double antibody sandwich ELISA method for detection of residual recombinant trypsin in influenza split virion vaccine(MDCK cells)
10.13200/j.cnki.cjb.004659
- VernacularTitle:流感病毒裂解疫苗（MDCK细胞）重组胰蛋白酶残留量双抗体夹心ELISA检测方法的验证
- Author:
Qingmei ZHANG
1
Author Information
1. Viral Vaccine Research and Development 2 Unit, Wuhan Institute of Biological Products Co., Ltd., Wuhan 430207, Hubei Province, China
- Publication Type:Journal Article
- Keywords:
Influenza split virion vaccine(MDCK cells);
Monovalent virus harvest fluid;
Monovalent bulk;
Residual recombinant trypsin(RT);
Double antibody sandwich ELISA
- From:
Chinese Journal of Biologicals
2026;39(02):174-179+188
- CountryChina
- Language:Chinese
-
Abstract:
Objective To verify the double antibody sandwich ELISA method for the detection of residual recombinant trypsin(RT) in monovalent virus harvest fluid and monovalent bulk solution of influenza split virion vaccine(MDCK cells).Methods Double antibody sandwich ELISA was used to detect residual levels of RT in monovalent virus harvest fluid and monovalent bulk solution, and was verified for the limit of quantitation(LOQ), specificity, repeatability, intermediate precision, accuracy and robustness.Results The method exhibited good linearity within the range of 10-0. 156 ng/mL, with a correlation coefficient R~2 of 1. 000. The LOQ was 0. 156 ng/mL. After diluting the monovalent harvest by a factor of two using sample diluent and phosphate buffer, the coefficient of variation(CV) for RT content was 1%, and the RT content measured after diluting monovalent bulk solution two-fold with sample diluent and phosphate buffer was less than 0. 156 ng/mL.The spiked recovery rates of RT solutions at different concentrations ranged from 81% to 105%, with a CV ranging from 1%to 6%. The CV of RT content measured by different test personnel in monovalent harvest was 3%, and the RT content in the monovalent bulk solution measured by different operators was also less than 0. 156 ng/mL. The CV of RT residue content in monovalent harvest at different chromogenic times was 5%, and the residual RT content in the monovalent bulk solution was less than 0. 156 ng/mL, indicating this detection method had good tolerance to different chromogenic times.Conclusion The double antibody sandwich ELISA method for detecting residual RT in monovalent virus harvest solution and monovalent bulk solution of influenza split virion vaccine(MDCK cells) has good linearity and range, LOQ, specificity, repeatability, intermediate precision, accuracy and robustness, and is suitable for the detection of residual RT in influenza split virion vaccine(MDCK cells).
