人免疫球蛋白中甲肝抗体效价ELISA检测方法的建立及验证

Development and verification of ELISA for hepatitis A antibody titer in human immunoglobulin

目的建立人免疫球蛋白产品中甲肝抗体效价ELISA检测方法,并对该方法进行验证及初步应用,以期满足《中国药典》三部(2015版)对人免疫球蛋白产品中甲肝抗体效价的质量控制检测要求。方法将人免疫球蛋白甲肝抗体国际标准品(97/646)稀释后,采用建立的ELISA法检测甲肝抗体效价,分别验证方法的线性、准确性和精密度,试验均重复3次。应用建立的方法检测我国3家企业共13批人免疫球蛋白产品中的甲肝抗体效价。结果甲肝抗体效价在10~50 m IU/ml浓度范围内线性良好,相关系数为0.994 8;用建立的方法检测浓度为15、25、35 m IU/ml人免疫球蛋白甲肝抗体国际标准品的回收率在86.9%~104.4%之间,3次测定结果的相对标准偏差(relative standard deviation,RSD)均小于10%。用建立的方法检测我国3家企业共13批人免疫球蛋白产品中的甲肝抗体效价,结果均低于《中国药典》三部(2015版)对甲肝抗体效价不低于100 IU/ml的新增要求。结论建立的ELISA方法快速、准确,可用于检测我国人免疫球蛋白产品的甲肝抗体效价。

Objective To develop, verify and preliminarily apply an ELISA method for determination of hepatitis A（HA）antibody tilter in human immunoglobulin. Methods The International Standard for Anti-Hepatitis A, Immunoglobulin（97/646）was diluted and determined for HA antibody titer by the developed ELISA method. The developed method was verified for linearity, accuracy and precision, each for 3 times. The HA antibody titers in 13 batches of human immuno-globulin from three manufacturers were determined by the method. Results The linear range of the developed method was 10 ~ 50 m IU/ml, with a correlation coefficient of 0. 994 8. The recovery rates of HA antibody titers in human immu-noglobulin at concentrations of 15, 25 and 35 m IU/ml were 86. 9% ~ 104. 4%, with RSDs of less than 10%. The HA antibody titers in 13 batches of human immunoglobulin from three manufacturers were not less than 100 IU/ml, which met the requirements in Chinese Pharmacopoeia（Volume Ⅲ, 2015 edition）. Conclusion The developed ELISA method was rapid and accurate, which might be used for the determination of HA antibody titer in human immunoglobulin in China.