Application of recombinant factor C method for bacterial endotoxin detection in recombinant SARS-CoV-2 vaccine(CHO cells)
10.13200/j.cnki.cjb.004629
- VernacularTitle:重组C因子法在重组新型冠状病毒疫苗（CHO细胞）细菌内毒素检测中的应用
- Author:
Hua LIU
1
Author Information
1. *Anhui Institute for Food and Drug Control, Key Laboratory of Public Health Social Governance, Philosophy and Social Sciences of Anhui Province, Recombinant Vaccine Research and Development Key Laboratory of Anhui Province Joint Construction, Hefei 230051, Anhui Province, China
- Publication Type:Journal Article
- Keywords:
Recombinant SARS-CoV-2 vaccine(CHO cells);
Recombinant factor C(rFC) method;
Endotoxin;
Fluorescence quantitative detection;
Limulus amebocyte lysate
- From:
Chinese Journal of Biologicals
2026;39(01):101-107
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the feasibility of recombinant factor C(rFC) method for quantitative detection of bacterial endotoxin in recombinant SARS-CoV-2 vaccine(CHO cells), and to provide more basis for the application of this method.Methods The standard curve of bacterial endotoxin was plotted by rFC method reagent, the dilution factor was determined through interference pre-experiment, the recovery rate of spiked endotoxin in the test solution was determined, and the quantitative detection test of bacterial endotoxin content in the test product was completed. The effect of aluminum adjuvant content on endotoxin detection was investigated, and the results were compared with those of the kinetic chromogenic assay and the gel clotting method described in current pharmacopeia.Results The linear range of the standard curve was 0. 05-50 EU/mL. The test sample diluted 50, 100 and 200 times did not interfere with the reaction, and the recovery rate of bacterial endotoxin fell in the range of 50% to 200%. The endotoxin quantitative test results of the three batches of test products all met the requirements, and were consistent with the gel clotting method. The Correl function correlation coefficient between aluminum adjuvant content an d endotoxin recovery rate determined by rFC method was-0. 828 3,showing a strong negative correlation.Conclusion The rFC method can be used for the quantitative detection of bacterial endotoxin in recombinant SARS-CoV-2 vaccine(CHO cells), with accurate results and high precision, and can reduce or even eradicate the usage of limulus resources, conforming to the 3R principle of world animal welfare.
