摘要
目的:分别用新型复合型层析介质Capto Core 400和传统介质Sepharose 6FF纯化Sabin株脊髓灰质炎病毒(Sabin strain poliovirus,sPV)去除宿主细胞蛋白(hostcell protein,HCP)和DNA,并且比较纯化结果及工艺参数。方法:用Capto Core 400与Sepharose 6FF纯化Vero细胞培养的Ⅰ、Ⅱ、Ⅲ型sPV浓缩液。分别检测纯化后sPV的D抗原含量、HCP残留量和DNA残留量,计算D抗原回收率、HCP和DNA去除率,并用t检验分析差异。结果:Capto Core 400与Sepharose 6FF纯化Ⅰ、Ⅱ、Ⅲ型sPV的D抗原回收率差异均无统计学意义(t值分别为1.09、1.08、1.02,P值均>0.05),但前者的Vero细胞HCP(t值分别为3.15、3.23、3.54)和DNA去除率均高于后者(t值分别为3.41、3.25、3.62),且差异有统计学意义(P值均<0.05)。Capto Core 400与Sepharose 6FF的介质使用体积比值为0.05,上样后纯化时间比值为0.04,工作缓冲液使用量比值为0.25,工艺线性流速比值为10,每升介质上样量比值为20,最大耐压力比值为2。结论:对比Sepharose 6FF,Capto Core 400可以更有效地去除Vero细胞HCP与DNA,各个工艺参数得到了优化,提高了sPV层析纯化的工作效率,并节约了时间和成本。
Objective To purify Sabin strain poliovirus(sPV)using new multimodal chromatography medium Capto Core 400 and traditional medium Sepharose 6FF to remove residual host cell protein(HCP)and DNA,comparing purification results and process parameters.Methods TypesⅠ,Ⅱ,ⅢsPV concentrates were purified using Capto Core 400 and Sepharose 6FF media,respectively.D antigen contents,HCP and DNA residues in purified sPV liquids were detected.D antigen recovery rates,as well as HCP and DNA removal rates were caculated and then analyzed by t-test.Results D antigen recovery rates of typesⅠ,Ⅱ,ⅢsPV purified by Capto Core 400 and Sepharose 6FF had no statistically significant difference(t values were 1.09,1.08,and 1.02,respectively,all P values>0.05).But HCP residue(t values were 3.15,3.23,and 3.54,respectively)and DNA residue removal rates(t values were 3.41,3.25,and 3.62,respectively)of the former were all higher than the latter,all with statistically significant differences(all P values<0.05).The specific values for Capto Core 400 and Sepharose 6FF process parameter ratios were:medium volume 0.05,purification time 0.04,application volume of buffer 0.25,process linear velocity 10,loading quantity of sample per liter medium 20,maximum withstand pressure 2.Conclusion Comparing with Sepharose 6FF,Capto Core 400 removes residual Vero cell HCP and DNA more effectively,with more highly optimized process parameters,resulting in increased sPV purification process efficiency and lowered time and cost.
作者
赵硕
彭博
孙浩
杜明磊
詹乐洋
张晋
李思越
张萌
梁宏阳
Zhao Shuo;Peng Bo;Sun Hao;Du Minglei;Zhan Leyang;Zhang Jin;Li Siyue;Zhang Meng;Liang Hongyang(The 5th Department of Vaccine,Beijing Institute of Biological Products Co.,Ltd.,Beijing 100176,China)
出处
《国际生物制品学杂志》
CAS
2020年第6期292-295,共4页
International Journal of Biologicals
