鼠疫耶尔森菌F1抗原制备中超滤工艺的优化  

Application and optimization of ultrafiltration process for Yersinia pestis F1 antigen

作　　者：李朋伟 王婷 孙田华 蒋保余 张光磊 马维民 焦磊 LI Pengwei;WANG Ting;SUN Tianhua;JIANG Baoyu;ZHANG Guanglei;MA Weimin;JIAO Lei(The Third Research Department,Lanzhou Institute of Biological Products Co.,Ltd.,Gansu Provincial Vaccine Technology Innovation Center,Lanzhou 730046,Gansu Province,China)

机构地区：[1]兰州生物制品研究所有限责任公司第三研究室甘肃省疫苗技术创新中心,甘肃兰州730046

出　　处：《微生物学免疫学进展》2024年第5期43-47,共5页Progress In Microbiology and Immunology

基　　金：国家重点研发计划(2018YFC1200600)。

摘　　要：目的通过单因素实验(experiment of single factor)和响应面分析法(response surface methodology)优化鼠疫耶尔森菌(Yersinia pestis)F1抗原纯化过程中的超滤工艺。方法以单因素试验优化鼠疫菌F1抗原超滤浓缩工艺,研究跨膜压、进液流速和料液质量浓度对膜通量的影响;采用Box-behnken中心设计法设计三因素两水平的响应面实验,建立二次回归模型,研究氨氮含量、料液质量浓度和跨膜压3个因素对F1抗原超滤过程中氨氮去除效率的影响,以优化鼠疫菌F1抗原超滤除盐工艺。结果单因素试验中,以进液速度300 mL/min和500 mL/min分别进行试验,跨膜压分别升至20.0 Psi和40.0 Psi后,膜通量不再明显增加;当料液蛋白浓度<0.30 g/L时,料液浓度升高不会对膜通量产生影响,料液蛋白浓度>0.30 g/L时,随着料液浓度升高,膜通量随之降低。响应面超滤除盐试验中,料液质量浓度和跨膜压对氨氮去除效率均有显著影响(P均<0.05),并且两者之间有交互作用。结论鼠疫菌F1抗原纯化过程中超滤工艺的应用,及其工艺条件的优化,可以显著节省F1抗原的纯化制备时间,同时也更易于生产规模的放大。Objective The ultrafiltration process of F1 antigen of Yersinia pestis was optimized by single factor experiments and response surface methodology.Methods Single factor experiment was used to optimize ultrafiltration concentration of F1 antigen of Yersinia pestis to study the effection of transmembrane pressure,feed flux,concentration of product on membrane flux.A response surface methodology based on Box-behnken design involving 3 factors at 2 levels was generated and the experimental data obtained were subjected to quadratic regression analysis to evaluate the effect of ammonia nitrogen concentration,product concentration,transmembrane pressure on ammonia nitrogen removal rate,in order to optimize the desalination process of Yersinia pestis F1 antigen ultrafiltration.Results In single factor experiments,the membrane flux no longer increased significantly after the transmembrane pressure was increased to 20.0 Psi and 40.0 Psi when experiments were conducted at feed flux of 300 mL/min and 500 mL/min,respectively.When the protein concentration in the feed solution was less than 0.30 g/L,an increase in the feed solution concentration would not affect the membrane flux.When the protein concentration in the feed solution was greater than 0.30 g/L,with the feed solution concentration increased,the membrane flux decreased.In the response surface ultrafiltration experiment,both product concentration and transmembrane pressure had a significant impact on the ammonia nitrogen removal efficiency,and there was an interaction between them.Conclusion The application of ultrafiltration technology and optimization of process conditions in the purification process of F1 antigen could significantly save the purification and preparation time of F1 antigen(P<0.05),and also make it easier to scale up production.

关 键 词：鼠疫耶尔森菌 F1抗原 超滤 响应面分析法 膜通量 超滤纯化 工艺优化 

分 类 号：R378.61[医药卫生—病原生物学]