腺病毒载体重组新型冠状病毒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 SARS-CoV-2 adenovirus vector-based vaccine potency test method in vitro

作　　者：苏歧 徐宏山[1] 王新竹 赵丹华 吴小红[1] 姜崴 叶强[1] 刘欣玉[1] SU Qi;XU Hongshan;WANG Xinzhu;ZHAO Danhua;WU Xiaohong;JIANG Wei;YE Qiang;LIU Xinyu(Division of Arboviral Vaccines,National Institutes for Food and Drug Control,Beijing 102629,China;Division of biotechnology,Changchun Institute of Biological Products Co.,Lid.,Changchun 30012,Jiling Province,China)

机构地区：[1]中国食品药品检定研究院虫媒病毒疫苗室,北京102629 [2]长春生物制品研究所有限责任公司生物技术室,吉林长春130012

出　　处：《微生物学免疫学进展》2024年第5期26-31,共6页Progress In Microbiology and Immunology

摘　　要：目的建立通用、稳定的腺病毒载体重组新型冠状病毒(severe acute respiratory syndrome 2,SARS-CoV-2)疫苗的体外生物学活性检测方法,并对其进行方法学验证。方法利用实验室之前筛选出的ELISA试剂盒筛选不同的细胞、细胞浓度和感染剂量,从而建立通用体外生物学活性检测方法并进行相对准确度、中间精密度、线性和范围的验证。结果建立了通用体外生物学活性检测方法,筛选出HEK293细胞为适宜感染用细胞,其铺板细胞浓度为8×10^(5)个/mL,细胞感染剂量是1.0×10^(7) VP。该方法经不同人员和日期检测验证,每个效价水平的相对偏倚应均在±30%内,水平效价测定值的几何变异系数(geometric coefficient of variation,GCV)≤30%,疫苗效价理论值(对数)与其相应的效价测定值(对数)作图并进行线性回归计算,直线回归方程的相关系数r≥0.95,效价水平范围在50%~200%,证明能满足对多种SARS-CoV-2变异株腺病毒载体疫苗的检定需要。结论建立了通用、稳定的腺病毒载体SARS-CoV-2疫苗体外生物学活性检测方法,该方法可应用于腺病毒载体SARS-CoV-2疫苗的质量控制。Objective To establish and verify a universal and stable potency test method in vitro for adenovirus vector-based Severe Acute Respiratory Syndrome Coronavirus 2(SARS-CoV-2)vaccines.Methods Different cells,cell concentrations and doses were screened using ELISA kits previously screened in the laboratory.And then a universal in vitro biological activity assay was established and relative accuracy,intermediate precision,linearity and it's range were validated.Results As the suitable cells for infection,HEK293 cell was laid on plate at the concentiation of 8×10^(5),the cellular infection dose was 1.0×10^(7) VP.The method was validated by diflerent personnel on different dates.The relative bias of each potency level should be within±30%,and the geometric coefficient of variation(GCV)of the horizontal potency measurements should be≤30%.The theoretical(logarithmic)values of the vaccine's potency were plotted against their corresponding potency measurements(logarithmic)and a linear regression was calculated.The correlation coefficients(r)of linear regression equations was≥0.95,and the potency levels ranged from 50%to 200%could meet the needs of the quality control.Conclusion A universal and stable method for potency test in vitro of SARS-CoV-2 adeno-virus vector-based vaccine has been established,which can be applied to the quality control of adenovirus vector-based vaccines.

关 键 词：重组新型冠状病毒疫苗(腺病毒载体) 质量控制 生物学活性 

分 类 号：Q-331[生物学]