摘要
目的利用层析技术制备高纯度的核酸疫苗。方法用层析技术中的凝胶过滤、亲和、离子交换层析3种方法，依次对核酸疫苗进行纯化，并检测其纯度。结果连续使用3种层析方法获得的DNA达到了理想的分离效果，琼脂糖凝胶电泳未检出RNA，A260与A280的比值介于1．80～2．00之间，其中环状DNA达90％以上。经离子交换层析后的内毒素含量小于10EU／mg DNA，宿主DNA残留量小于0．002μg/μg DNA。结论纯化的核酸疫苗均符合国家标准，为制备核酸疫苗奠定了基础。
Objective To prepare highly purified DNA by chromatography. Methods Purify DNA vaccine by gel fdtration, affinity and ion exchange chromatography in turn, and determined the purity of purified vaccine. Results After purification by 3 kinds of chromatography,no RNA was detected in DNA vaccine by agarese gel electrophoresis. The ratio of A260 to A280 was 1. 80-2. 00. The content of circular DNA was more than 90%. The endotoxin and host DNA contents were less than 10 EU/mg DNA and less than 0. 002 μg/μg DNA respectively. Conclusion The purified DNA vaccine met the relevant national requirements.
出处
《中国生物制品学杂志》
CAS
CSCD
2007年第2期129-132,共4页
Chinese Journal of Biologicals
基金
国家863基金资助项目(2005AA218030)
关键词
核酸疫苗
层析技术
纯化
DNA vaccine
Chromatography
Puritification
