SARS-CoV-2mRNA疫苗通用体外生物学活性检测方法的建立及验证
Establishment and validation of a universal test method in vitro for SARS-CoV-2 mRNA vaccine potency
王新竹 1赵丹华 2苏歧 1刘欣玉 2姜崴 3叶强2
作者信息
- 1. 中国食品药品检定研究院,北京 102629;长春生物制品研究所有限责任公司,吉林长春 130012
- 2. 中国食品药品检定研究院,北京 102629
- 3. 长春生物制品研究所有限责任公司,吉林长春 130012
- 折叠
摘要
目的 建立通用稳定的SARS-CoV-2 mRNA疫苗的体外生物学活性检测方法,并对方法进行验证,以期用于SARS-CoV-2 mRNA疫苗的质量控制.方法 筛选能够对SARS-CoV-2各变异株S蛋白均能较好结合的ELISA试剂盒及转染细胞、细胞铺板浓度、转染剂量,建立SARS-CoV-2 mRNA疫苗体外生物学活性检测方法,并进行验证.结果 筛选到1种对各变异株S蛋白均能良好结合的试剂盒,并确定了转染细胞为HEK293T、细胞铺板浓度为2.5 × 105个/mL、转染剂量为6孔板4μg/孔,建立了通用稳定的SARS-CoV-2 mRNA疫苗体外生物学活性检测方法.验证结果显示,该方法能满足检定需求.结论 建立的SARS-CoV-2mRNA疫苗体外生物学活性检测方法相对准确度、线性、中间精密度、范围良好,可用于SARS-CoV-2 mRNA疫苗的质量控制.
Abstract
Objective To establish and verify a universal and stable potency test method in vitro for SARS-CoV-2 mRNA vaccine,so as to use it for the quality control of SARS-CoV-2 mRNA vaccine.Methods ELISA kits that could bind well to S protein of SARS-CoV-2 variants,as well as transfected cells,cell plating concentrations and doses for transfection were screened,and then a potency test method for SARS-CoV-2 mRNA vaccine in vitro was established and verified.Results An ELISA kit was found with good binding ability to S protein of each variant,and HEK293T cells were determined as the transfection cells,with the plating concentration of 2.5 x 105 cells/mL and the transfection dose of 4 μg/well in the 6-well plate.An universal and stable potency test method for SARS-CoV-2 mRNA vaccine in vitro was established.The verification results showed that the method met the quality control needs.Conclusion The established potency test method in vitro for SARS-CoV-2 mRNA vaccine has good relative accuracy,linearity,intermediate precision and range,and can be applied to the quality control of SARS-CoV-2 mRNA vaccines.
关键词
SARS-CoV-2/mRNA疫苗/体外生物学活性/酶联免疫吸附测定Key words
SARS-CoV-2/mRNA vaccine/Potency in vitro/Enzyme-linked immunosorbent assay(ELISA)引用本文复制引用
基金项目
国家重点研发计划(2020YFC0860500)
出版年
2024
NETL
NSTL
万方数据