降纤酶制备中层析工艺对病毒去除能力研究

Study on the Virus Removal Ability of Chromatographic Process in the Preparation of Defibrase

目的:研究降纤酶制备中层析工艺对脂包膜指示病毒伪狂犬病毒(PRV)和水疱性口腔炎病毒(VSV)的去除能力,为该产品的病毒安全性评价提供依据。方法:用微量滴定法测定层析工艺处理前后各批次样品中PRV和VSV的滴度,按Karber法计算病毒滴度,并计算层析处理前后各批次样品中各指示病毒的滴度下降值。结果:层析填料(使用≥108次)对PRV的去除效果为4.28 logs,层析填料(使用≥102次)对VSV的去除效果为4.72 logs;未处理对照中,PRV滴度平均下降0.14 logs、VSV滴度平均下降0.32 logs。结论:降纤酶制备工艺中的层析工艺能有效去除脂包膜指示病毒。

Objective:To study the capabilities of removing lipid-envelope indicator viruses(PRV and VSV)by the chromatography process in the preparation of defibrase in order to lay a basis for the virus safety evaluation of the products.Methods:The titers of PRV and VSV in each batch of samples before and after the chromatographic process were measured by microtitration method.The virus titers were calculated with the Karber method,and the titer drop values of each indicator virus(PRV and VSV)in each batch of samples before and after the chromatographic treatment were calculated.Results:The removal effect of chromatography packing(used≥108 times)on PRV was 4.28 logs,and the removal effect of chromatography packing(used≥102 times)on VSV was 4.72 logs.The PRV titer in the untreated control has decreased by 0.14 logs and VSV titer has decreased by 0.32 logs on average.Conclusion:The chromatographic in the preparation of defibrase can effectively remove the lipid-envelope indicator virus.