重组SARS-CoV-2 S1蛋白聚集对抗原效价的影响
10.13200/j.cnki.cjb.004723
- VernacularTitle:Effect of aggregation of recombinant SARS-CoV-2 S1 protein on antigenic potency
- Author:
Qiuping ZOU
- Publication Type:Journal Article
- Keywords:
SARS-CoV-2 vaccine;
Recombinant S1 protein;
Protein aggregation;
Antigenic potency
- From:
Chinese Journal of Biologicals
2026;39(06):659-666
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of the aggregate content of recombinant SARS-CoV-2 S1 protein on its antigenic potency,and to provide technical reference for the quality control of S1 protein as a substrate in SARS-CoV-2 vaccine development.Methods A double-antibody sandwich ELISA was employed to establish an in vitro antigenic potency measurement system for the S1 protein of the SARS-CoV-2 JN.1 variant.The impact of aggregation during the manufacturing process on the evaluation of both in vitro antigen potency and in vivo immunogenicity was analyzed.Results After heating denaturation,S1 protein was completely denatured(Tm = 55.5 ℃),and the antigen titer decreased significantly from(1.5 ±0.04) JEU/μg to(0.3 ± 0.02) JEU/μg(t_4= 46.47,P < 0.000 1).The protein was not degraded but aggregates appeared.Under non-denaturing conditions,molecular exclusion chromatography obtained four components(Ⅰ-Ⅳ,the proportion of aggregates decreased in turn) with a gradient of aggregates ratio.The antigen titer of component Ⅰ(aggregates > 95%) with the highest aggregates content was(16.8 ± 0.98) JEU/μg,which was about 13 times that of monomer component Ⅳ[aggregates < 4%,titer(1.3 ± 0.03) JEU/μg].Components with a higher proportion of aggregates and higher in vitro potency induced higher antibody titers,and there was a significant positive correlation between in vitro antigenic potency and in vivo antigenicity.Conclusion The proportion of SARS-CoV-2 S1 protein aggregates can serve as a key indicator for optimizing the manufacturing process and evaluating the quality and stability of bulk solutions and preparations.
