DEAE-Sepharose FF纯化脊髓灰质炎病毒Sabin株Ⅰ型稳定性研究  

Study on the stability of DEAE-Sepharose FF purified poliovirus type 1 Sabin strain

作　　者：高承刚 李卫东 杨晓蕾 GAO Chenggang;LI Weidong;YANG Xiaolei(Institute of Medical Biology, Chinese Academy of Medical Science and Peking Union Medical College , Kunming 650018, China)

机构地区：[1]中国医学科学院·北京协和医学院·医学生物学研究所

出　　处：《西部医学》2019年第6期831-835,共5页Medical Journal of West China

基　　金：国家科技部重大专项(2013ZX10004003-003-002);国际合作项目(2011DFR30420);中国医学科学院医学科技创新团队(2016-I2M-3-026);云南省创新团队(2015HC027)

摘　　要：目的观察离子交换纯化脊髓灰质炎病毒Ⅰ型Sabin株稳定性,以研究病毒纯化的回收率和质量控制。方法使用微载体培养Vero细胞和脊灰病毒Ⅰ型Sabin株,经过逐级过滤澄清和100K超滤膜浓缩,再经凝胶介质初步纯化后的病毒液,然后观察经过离子介质DEAE Sepharose FF层析纯化5批次的纯化效果。对每次纯化获得的病毒液测定D抗原、蛋白质含量、牛血清白蛋白含量、DNA残留量以评价其质量。结果凝胶纯化后的病毒液,经离子介质DEAE Sepharose FF纯化,其D抗原、蛋白质含量、牛血清白蛋白含量、DNA残留量和比活性的批间差异不显著(P>0.05)。结论离子介质DEAE Sepharose FF在一定批次内纯化脊灰病毒工艺稳定。Objective To study the recovery efficiency and quality control system for poliovirus Type Ⅰ Sabin strain, purified by ion exchange. Methods Vero cells were seeded on the microcarrier, infected by poliovirus type Ⅰ Sabin strains. After harvesting of virus, the systematic purification process was taken, including multistage filtration clarification, 100K ultrafiltration membrane and then gel medium. The purification efficiency of ion medium DEAE Sepharose FF chromatography was observed for 5 batches. The key quality control points including D antigen, protein content, bovine serum albumin content and DNA residue were monitored and evaluated for each times. Results After purified poliovirus Type Ⅰ Sabin strain by DEAE Sepharose FF, there was no significant difference in D antigen, protein content, bovine serum albumin content, DNA residue and specific activity between each batch. Conclusion The purification method of DEAE Sepharose FF for poliovirus Type Ⅰ Sabin strain was useful and stability.

关 键 词：脊髓灰质炎病毒 DEAE SEPHAROSE FF 介质稳定性 

分 类 号：R512.4[医药卫生—内科学]