流感病毒超滤工艺的优化  

Optimization of process for ultrafiltration of influenza virus

作　　者：高菁霞[1] 宋绍辉[1] 王东宝[1] 杨伟[1] 欧阳圣洁 代小虎[1] 马磊[1] 廖国阳[1] 李卫东[1] GAO Jing-xia;SONG Shao-hui;WANG Dong-bao;YANG Wei;OUYANG Sheng-jie;DAI Xiao-hu;MA Lei;LIAO Guo-yang;LI Wei-dong(Institute of Medical Biology, Chinese Academy of Medical Science, Kunming 650118, Yunnan Province, China)

机构地区：[1]中国医学科学院医学生物学研究所,云南昆明650118

出　　处：《中国生物制品学杂志》2018年第6期662-666,共5页Chinese Journal of Biologicals

基　　金：北京协和医学院中国医学科学院重大协同创新项目创新工程四价流感病毒裂解疫苗(2016-I2M-1-019);北京协和医学院中国医学科学院医学与健康科技创新工程协同创新团队项目(2016-I2M-3-026);云南省科技创新人才计划项目中国医学科学院医学生物学研究所疫苗研发与新技术应用研究省创新团队项目(2015HC027)

摘　　要：目的分析截留相对分子质量不同的超滤膜浓缩对流感病毒纯化效果的影响,以获得杂质含量低的高浓度病毒液。方法将H1N1、H3N2、Bv和By 4个型别流感病毒分别接种9~11日龄鸡胚,经(34±1)℃培养72 h后,收获尿囊液,经连续流离心获得澄清的病毒液后,再进行截留相对分子质量1 000 000、300 000的超滤膜及其两种超滤膜组合的超滤浓缩,测定超滤前后的总蛋白含量、卵清蛋白含量和血凝滴度。结果截留相对分子质量1 000 000的超滤膜超滤后,杂质去除率高,均在80%以上,截留相对分子质量300 000的超滤膜超滤后杂质去除率相对低,但后者病毒回收率较高,二者组合既能降低杂蛋白含量,又可提高病毒回收率。结论流感病毒鸡胚尿囊液采用截留相对分子质量1 000 000+300 000超滤膜组合超滤浓缩,病毒纯化效果较好。Objective To analyze the effect of concentration by ultrafiltration membrane with various relative molecular mass cutoffs on purification of influenza virus. Methods Four types of influenza viruses（H1 N1,H3 N2,Bv and By）were inoculated into 9-to 11-day-old chick embryo respectively,and cultured at（34 ± 1）℃ for 72 h. The allantoic fluid was harvested,clarified by continuous flow centrifugation,and concentrated by using the ultrafiltration membranes with relative molecular mass cutoffs of 1 000 000 and 300 000 alone or in combination. The total protein and ovalbumin contents and hemagglutination titer before and after ultrafiltration were determined. Results The removal efficiency of foreign matters by ultrafiltration using membrane with relative molecular mass cutoff of 1 000 000 was more than 80%.However,the removal efficiency by the membrane with relative molecular mass cutoff of 300 000 was relatively low,while the virus recovery rate was relatively high. The combination of two methods decreased the foreign protein content and increased the virus recovery rate. Conclusion Influenza virus was effectively purified by concentration of chick embryo allantoic fluid by the combination of ultrafiltration membranes with relative molecular mass cutoffs of 1 000 000 and 300 000.

关 键 词：流感病毒 鸡胚尿囊液 超滤 纯度 回收率 

分 类 号：R373.13[医药卫生—病原生物学] R392-33[医药卫生—基础医学]