破伤风人免疫球蛋白抗-HBs效价测定(ELISA法)方法验证  

Human Tetanus Immunoglobulin Anti-HBS Determination(ELISA)Method Validation

作　　者：吴翠 纪胜榆 周婷 WU Cui;JI Shengyu;ZHOU Ting(Guangdong Well Biological Pharmaceutical Co.,Ltd.,Shantou 515071,China)

机构地区：[1]广东卫伦生物制药有限公司,广东汕头515071

出　　处：《生物化工》2024年第2期123-126,共4页Biological Chemical Engineering

摘　　要：目的:对ELISA法测定破伤风人免疫球蛋白抗-HBs效价进行方法学验证。方法:根据《中国药典》(2020年版)“9401生物制品生物活性/效价测定方法验证指导原则”,对破伤风人免疫球蛋白抗-HBs效价测定(ELISA法)方法进行专属性、相对准确度、中间精密度、线性及线性范围的验证,并与放射免疫方法(RIA法)测定结果进行比对。结果:专属性验证中,回收率在88%~110%,实测与理论结果等效,样品、混合样品均与标准回归曲线平行;相对准确度验证中,相对偏倚在-6.9%~5.0%,回归曲线斜率0.9981;中间精密度验证中,几何变异系数<10%;线性验证中,相关系数为0.9970;方法范围为10~120 mIU/mL。ELISA法测得的每1 g蛋白质中抗-HBs结果平均值比RIA法低0.5 IU。结论:经验证,ELISA法可替代RIA法用于破伤风人免疫球蛋白中抗-HBs效价的测定。Objective:To validate the method for the determination of anti-HBS titer of human tetanus immunoglobulin by ELISA.Methods:According to the validation guidelines of 9401 biological activity/titer determination method of biological products in Chinese Pharmacopoeia(2020 edition),the specificity,relative accuracy,intermediate precision,linear and linear range of human tetanus immunoglobulin anti-HBS assay(ELISA)method are verified.The results are compared with those of radioimmunoassay(RIA).Results:In the specific verification,the recoveries are between 88%and 110%,the theoretical and measured results are equivalent,and the samples and mixed samples are parallel to the standard regression curve;in the verification of relative accuracy,relative bias ranges from-6.9%to 5.0%,and the slope of regression curve is 0.9981;in intermediate precision verification,geometric coefficient of variation are all<10%;in linear verification,the correlation coefficient of regression curve is 0.9970;method range is 10 mIU/mL to 120 mIU/mL.The average value of anti-HBS 6 times per 1 g of protein measured by ELISA is 0.5 IU lower than that by RIA.Conclusion:It is proved that ELISA can replace RIA for the determination of anti-HBS titer in human tetanus immunoglobulin.

关 键 词：抗-HBs效价测定 ELISA法 破伤风人免疫球蛋白 

分 类 号：O657.7[理学—分析化学]