论文部分内容阅读
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(P>0.05);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。
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 analyze the clustering activity of pertussis toxin (PT) in CHO cells from different sources. Guinea pigs were randomly divided into experimental group of imported vaccine Lot 1 ~ 3, domestic vaccine Lot 4 experimental group, reference vaccine control group, at the same time set unimmunized control group. On days 0 and 21, respectively, 0.5 ml of the vaccine was injected subcutaneously into the abdomen of guinea pigs. On the 28th day after initial immunization, the blood was collected from the heart and the serum was separated. The titer of PT antibody was detected by ELISA, and the titer of neutralizing antibody was detected by the CHO cell cluster test. Results There was a certain difference in the ability of PT to collect CHO cells from different origins, among which the CHO cells from ATCC were the most sensitive. Each acellular pertussis vaccine can induce better PT antibody immune response in animal models of guinea pigs, compared with imported vaccines and domestic vaccines, the difference was not statistically significant (P> 0.05); The results of two methods Has a certain relevance, Spearman correlation coefficient of 0.474. Conclusions The applicability of the CHO cell cluster assay in quality control and PT immunogenicity analysis during the purification of pertussis vaccine was confirmed, which lays the foundation for strengthening and improving the quality control of acellular pertussis vaccine in China.