层析法去除A群流脑多糖疫苗粗糖中杂蛋白

A Chromotography Method for Removing Impurity Proteins from Meningococcal Polysaccharide Type A Sample

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.