CHO细胞簇集试验在无细胞百日咳疫苗质量控制中的应用

发布时间：2017-12-27 03:07

本文关键词：CHO细胞簇集试验在无细胞百日咳疫苗质量控制中的应用　出处：《中国生物制品学杂志》2014年10期 　论文类型：期刊论文

【摘要】：目的探讨CHO细胞簇集试验在无细胞百日咳疫苗质量控制中的应用。方法采用CHO细胞簇集试验比较分析不同来源的CHO细胞对百日咳毒素(pertussis toxin,PT)簇集活性。将豚鼠随机分为进口疫苗Lot 1~3实验组、国产疫苗Lot 4实验组、参考疫苗对照组,同时设未免疫对照组。分别于第0和21天,经豚鼠腹部皮下注射0.5 ml疫苗。于初免后第28天,经心脏采血,分离血清,采用ELISA法检测PT抗体滴度,CHO细胞簇集试验检测中和抗体滴度。结果 PT对不同来源的CHO细胞簇集能力存在一定差异,其中以来自ATCC的CHO细胞的簇集最敏感。各无细胞百日咳疫苗在豚鼠动物模型上均能诱导较好的PT抗体免疫反应,进口疫苗与参考疫苗及国产疫苗相比,差异均无统计学意义(P0.05);两种方法的检测结果具有一定的相关性,Spearman相关系数为0.474。结论证实了CHO细胞簇集试验用于百日咳疫苗纯化过程中质量控制和PT免疫原性分析的适用性,为加强和完善我国无细胞百日咳疫苗的质量控制奠定了基础。
[Abstract]:Objective to investigate the application of CHO cell cluster test in the quality control of acellular pertussis vaccine. Methods CHO cell cluster test was used to compare and analyze the cluster activity of CHO cells from different sources to pertussis toxin (pertussis toxin, PT). The guinea pigs were randomly divided into the imported vaccine Lot 1~3 experimental group, the domestic vaccine Lot 4 experimental group, the reference vaccine control group, and the unimmunized control group. At zeroth and twenty-first days, 0.5 ml vaccine was injected subcutaneously into the abdomen of the guinea pig. Twenty-eighth days after primary exemption, blood was collected from the heart and serum was separated. The titer of PT antibody was detected by ELISA, and the titer of neutralizing antibody was detected by CHO cell cluster test. Results there was a certain difference in the clustering ability of CHO cells from different sources, in which the cluster of CHO cells from ATCC was the most sensitive. The acellular pertussis vaccine could induce PT immune response as well as in the animal model of guinea pig, compared with imported vaccines and reference vaccine and domestic vaccine, there were no significant differences (P0.05); the test results of two methods had a certain correlation, Spearman correlation coefficient is 0.474. Conclusion the applicability of CHO cell cluster assay in quality control and PT immunogenicity analysis of pertussis vaccine purification process is confirmed, which lays the foundation for strengthening and improving the quality control of acellular pertussis vaccine in China.
【作者单位】：中国食品药品检定研究院卫生部生物技术产品检定方法及其标准化重点实验室;
【基金】：国家863计划“疫苗效果和质量评价新技术研究”(2012AA02A402)
【分类号】：R378.99;R392-33
【正文快照】： 百日咳毒素(pertussis toxin,PT)是百日咳致病机制中起主要作用的毒力因子,同时也是百日咳杆菌产生的诸多生物活性物质中唯一不受争议的保护性抗原。PT为目前无细胞百日咳疫苗中最主要的组分[1]。因此,在疫苗生产过程中,PT脱毒工艺监控和免疫原性分析一直是疫苗质量控制的重点

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