摘要
目的:建立用层析法去除脑脊髓膜炎球菌A群荚膜多糖样品中的杂蛋白的方法,以替代传统的苯酚抽提。方法:A群流脑多糖粗糖经过新型多模式离子交换填料Capto adhere,使用低盐上样高盐洗脱,洗脱的目的峰再经过Sephardex G-25凝胶过滤脱盐换成水,即为流脑多糖精糖原液。结果:采用上述方法制备的精制多糖样品中杂蛋白占固总的比例低于0.2%,符合WHO现行规程中的杂蛋白比例低于1%的要求,核酸、内毒素等生化指标均符合现行规程,整个层析过程的回收率大于65%。结论:层析法去除A群流脑多糖粗糖中的杂蛋白是有效的,比传统的苯酚抽提的方法更有利于环保,易于操作,易于放大。
Objective：Set a new chromatography method to remove impurity proteins from meningococcal polysaccharide type A sample instead of the traditional phenol abstraction method. Method：This new method includes two purification steps by using Capto adhere and Sephardex G-25. Result：The contaminative protein concentration is 0.2% in final meningococcal polysaccharise type A sample, which is below the WHO regulation standard （ 1% ）, the content of nucleic acid and endotoxin are also blow the current regulation standard, the recovery is over 65%. Conclusion：Compare with the tradition phenol abstraction method, the chromatography method can remove impurity proteins effectively with high recovery and reducing environment pollution and easy to scale up.
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2010年第8期88-92,共5页
China Biotechnology
