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层析技术纯化核酸疫苗
Purification of DNA Vaccine by Chromatography
【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。
【Abstract】 Objective To prepare highly purified DNA by chromatography.Methods Purify DNA vaccine by gel filtration,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 agarose 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.
- 【文献出处】 中国生物制品学杂志 ,Chinese Journal of Biologicals , 编辑部邮箱 ,2007年02期
- 【分类号】R392
- 【被引频次】7
- 【下载频次】596