原代地鼠肾细胞疫苗中残余宿主细胞蛋白检测方法的建立及初步验证

Establishment and initial validation of an ELISA method for determination of host cell protein in PHKC vaccines

目的建立定量检测原代地鼠肾细胞（primary hamster kidney cell，PHKC）疫苗中残余宿主细胞蛋白含量的方法。方法制备PHKC蛋白免疫家兔，对获得的抗血清进行纯化并标记辣根过氧化物酶，通过优化各反应条件建立ELISA方法，同时进行方法学验证。结果ELISA检测方法的最佳包被抗体质量浓度为6mg／L，酶标抗体工作浓度为1：2000，最佳检测区间为12．500～200．000μg／L，定量限度为23．250μg／L，准确度和精密度较佳。结论建立了一种简单、准确、可靠检测PHKC病毒性疫苗中宿主细胞蛋白残留量的ELISA双抗体夹心法。

Objective To establish an ELISA method for quantitative determination of the residual primary hamster kidney cell （PHKC） proteins in vaccines. Methods The PHKC proteins were prepared and used to immunize rabbits. The antisera were purified and labeled with horseradish peroxidase. The reaction conditions of the ELISA method were optimized in various aspects and the method was validated by a series of tests. Results The optimal coating antibody concentration was 6 mg/L, and the dilution of the enzyme-labeled antibody was 1 ： 2000. The best linear range was 12. 500-200. 000 μg/L, and the quantitation limit was 23. 250 μg/L. The ELISA method had good precision and accuracy. Conclusion The double antibody sandwich ELISA method which is easy, accuarte and reliable for quantitative determination of the residual PHKC proteins in vaccines is established.