Sabin株脊髓灰质炎病毒纯化的离子交换层析条件研究

目的  研究Sabin株脊髓灰质炎病毒（Sabin strain polio virus，sPV）纯化的离子交换层析（ion exchange chromatography，IEC）条件。 方法  在不同层析条件下对sPV凝胶层析粗纯液进行IEC，设定的层析参数分别为介质Q Sepharose FF或Eshmuno Q、上样量30%或40%柱体积、流速（150±5）或（240±8） cm/h。分别检测各型sPV纯化液的D抗原含量、蛋白浓度、病毒滴度、宿主细胞蛋白残留量和Vero细胞DNA残留量，计算D抗原回收率和比活。结果  Eshmuno Q IEC纯化的各型sPV纯化液的D抗原回收率均明显高于Q Sepharose FF IEC。在上样量40% 柱体积、流速（150±5） cm/h条件下，Eshmuno Q IEC纯化的各型sPV纯化液的D抗原比活均高于Q Sepharose FF IEC，差异均有统计学意义（Ⅰ型：t=4.23，Ⅱ型：t=5.73，Ⅲ型：t=4.18，P值均<0.05），而且前者的宿主细胞蛋白残留量（Ⅰ型：t=8.29，Ⅱ型：t=7.89，Ⅲ型：t=8.18，P值均<0.05）和Vero细胞DNA残留量（Ⅰ型：t=4.23，Ⅱ型：t=4.56，Ⅲ型：t=4.78，P值均<0.05）均低于后者，差异均有统计学意义。结论  用IEC纯化sPV进行纯化时，以Eshmuno Q代替Q Sepharose FF可增加上样量和流速，从而缩短IEC时间，提高纯化效率。

Study on ion exchange chromatography condition for purification of Sabin strain polio virus

Objective   To study on ion exchange chromatography (IEC) condition for purification of Sabin strain polio virus (sPV).  Methods  The crude purified liquids of sPV obtained by gel chromatography were subjected to IEC in different conditions, and setting parameters included chromatography medium Q Sepharose FF or Eshmuno Q, loading sample volume 30% or 40% column   volume (CV), and flow velocity (150±5) or (240±8) cm/h. D antigen contents, protein concenrations, virus titres, host cell protein（HCP）residues and Vero cell DNA residues in purified liquids of sPV purified by IEC were detected, and the recovery rates and specific activities of D antigen were caculated.  Results  The D antigen recovery rates of the Eshmuno Q IEC purified liquids of sPV were all higher than those of Q Sepharose FF IEC purified. In the conditions of loading sample volume 40% CV and flow velocity (150±5) cm/h, the D antigen specific activities of the Eshmuno Q IEC purified liquids of sPV were all higher than those of Q Sepharose FF IEC purified, and the differences were all statistically significant (typeⅠ: t=4.23, typeⅡ: t=5.73, type Ⅲ: t=4.18,  all P values <0.05), and HCP residues (typeⅠ: t=8.29, typeⅡ: t=7.89, type Ⅲ: t=8.18, all P values <0.05) and Vero cell DNA residues (typeⅠ: t=4.23, typeⅡ: t=4.56, type Ⅲ: t=4.78, all P values  <0.05) of the former were all lower than those of the latter, all with statistically significant differences. Conclusion  When sPV is purified with IEC, replacing Q Sepharose FF with Eshmuno Q can increase loading sample volume and flow velocity of IEC, thus the time of IEC is shortened and the purification efficiency is improved.