牛结核杆菌DNA疫苗的纯化工艺研究及质量鉴定

为获得纯度较高,能进行动物免疫试验的DNA疫苗,在高密度发酵工程菌的基础上,通过传统的碱裂解,获得粗提的质粒;用氯化钙去除裂解液中的大部分RNA,并用Sartobran P囊式过滤器获得了澄清的溶液。用聚乙二醇法沉淀超螺旋质粒DNA,去除大部分菌体蛋白和宿主DNA。最后用source 30Q阴离子交换色谱填料去除几乎全部的内毒素并浓缩了质粒。本试验纯化的DNA疫苗经质量检验达到了转染动物的要求。该工艺实现了一步色谱纯化DNA疫苗的目的,具有工艺简单、快速的优点。

Study the Preparation Technology and Quality Evaluation of 1Mycobacterium Bovis DNA Vaccine

In order to get high purity plasmids DNA which can be used for animal immunity,on the base of high density fermentive engineering bacteria,plasmids was isolated rudely through conventional alkaline lysis. Major RNA in the alkaline lysate was removed by calcium chloridize,and clarified solution was obtained by Sartobran P bladder-type filter; plasmid supercoil DNA was precipitated,and major bacteria proteins and host genomic DNA were removed by polyethyene glycol method. Finally,the quality monitoring of the DNA vaccine was accomplished the requirement for transfection of animal.This technique achieved one-step chromatographic purification of DNA vaccine,with the advantages of simplicity and rapidity.