摘要
目的对重组融合蛋白柱层析病毒去除工艺进行验证。方法以脑心肌炎病毒(encephalomyocarditis virus,EMCV)和猪细小病毒(porcine parvovirus,PPV)为指示病毒,考察重组融合蛋白生产中柱层析步骤(染料亲和层析、阴离子交换层析和凝胶层析)去除病毒的效果。结果经染料亲和层析、阴离子交换层析、凝胶层析后,EMCV滴度降低平均值分别为1.968、1.984和4.391 log10,3步柱层析共去除EMCV 8.343 log10;PPV滴度降低平均值分别为2.135、3.936和2.048 log10,3步柱层析共去除PPV 8.119 log10。结论染料亲和层析、阴离子交换层析、凝胶层析3步柱层析联合处理,在生产过程中可有效去除指示病毒(EMCV和PPV)及其所代表的相关病毒。
Objective To verify the procedure for virus removal from recombinant fusion protein by column chromatography. Methods The efficacies of virus removal from recombinant fusion protein by column, including dye affinity chromatography,anion exchange chromatography and gel filtration chromatography,were evaluated by using ence- phalomyocarditis virus（EMCV） and porcine parvovirus（PPV） as indicators. Results The EMCV titer decreased by 1. 968,1. 984 and 4. 391 log10 after purification by dye affinity chromatography,anion exchange chromatography and gel filtration chromatography,while the PPV titer decreased by 2. 135,3. 936 and 2. 048 log10,respectively. However,the to tal removals of EMCV and PPV after the three steps of chromatography were 8. 343 and 8. 119 log10 respectively. Conclusion EMCV and PPV as indicators as well as the relevant viruses may be removed effectively by combination of dye affinity chromatography,anion exchange chromatography and gel filtration chromatography.
出处
《中国生物制品学杂志》
CAS
CSCD
2014年第2期241-244,共4页
Chinese Journal of Biologicals
基金
中国食品药品检定研究院中青年基金项目(2011C4)
关键词
重组融合蛋白
柱层析
病毒去除
Recombinant fusion protein
Column chromatography
Virus removal
