国产层析填料在百日咳毒素和丝状血凝素纯化中的筛选及应用  

Screening and application of domestic chromatography resin in purification of pertussis toxin and filamentous hemagglutinin

作　　者：付慧 王玲 张娜 赵芳圆 何鹏 李世慧 FU Hui;WANG Ling;ZHANG Na;ZHAO Fang-yuan;HE Peng;LI Shi-hui(Bacterial vaccine Department of Research,Beijing Biological Products Institute Co.,Ltd.,Beijing 100176,China)

机构地区：[1]北京生物制品研究所有限责任公司细菌类疫苗研究室,北京100176

出　　处：《微生物学免疫学进展》2023年第3期36-44,共9页Progress In Microbiology and Immunology

基　　金：国家科技重大专项“重大新药创制”(2018ZX09738003⁃001)。

摘　　要：目的筛选适合的国产层析填料以满足对组分百日咳疫苗中百日咳毒素(pertussis toxin,PT)与丝状血凝素(filamentous hemagglutinin,FHA)的分离纯化。方法筛选对PT与FHA结合能力较好且分离度高的国产层析填料,优化纯化工艺,通过3批纯化试验比较国产填料与进口填料的抗原回收率、目的抗原纯度以及对目的抗原的动态结合载量。结果通过纯化试验筛选到国产填料SP resin-1与MMC resin-1并优化了纯化工艺。3批纯化试验显示,SP resin-1纯化百日咳抗原工艺稳定,FHA的纯度和回收率与Capto SP ImpRes差异无统计学意义(P>0.05);且与Capto SP ImpRes相比,SP resin-1对抗原的动态结合载量更高。3批PT精纯试验显示,国产填料MMC resin-1精纯PT的工艺稳定,PT的纯度和回收率均与Capto MMC差异无统计学意义(P>0.05),并且MMC resin-1对PT的载量高于Capto MMC。结论试验筛选出纯化PT与FHA的国产填料SP resin-1与MMC resin-1,纯化结果稳定,且纯化的抗原回收率与抗原纯度非劣效于进口填料。Objective To screen suitable domestic chromatographic resins for the separation and purification of pertussis toxin(PT)and filamentous hemagglutinin(FHA)from fractionated pertussis vaccine.Methods We screened domestic chromatographic resin with good binding ability and high separation of PT and FHA,optimized the purification process,and compared the antigen recovery,target antigen purity and dynamic binding load of the target antigen between domestic resin and imported resin through three batches of purification tests.Results The purification process of pertussis antigen with SP resin-1 and MMC resin-1 was stable,and the differences in the purity and recovery of FHA were not statistically significant compared with Capto SP ImpRes(P>0.05);and compared with Capto SP ImpRes Three batches of PT purification tests showed that the process of purifying PT with the domestic MMC resin-1 was stable,the purity and recovery of PT were not statistically different from those of Capto MMC(P>0.05),and the loading of PT with MMC resin-1 was higher than that of Capto MMC.Conclusion The domestic SP resin-1 and MMC resin-1 were screened out for the purification of PT and FHA.The purification results were stable,the antigen recovery and purity were not inferior to those of imported resins.

关 键 词：百日咳纯化 层析填料 百日咳毒素 丝状血凝素 

分 类 号：R378[医药卫生—病原生物学]