W_(135)和Y群脑膜炎球菌培养和多糖纯化

目的培养W135和Y群脑膜炎球菌及荚膜多糖的纯化。方法采用综合培养基代替半综合培养基在75L生物反应器中对两群脑膜炎球菌进行培养,研究培养基处方、发酵、多糖提取及纯化工艺。结果综合培养基可代替半综合培养基对两群脑膜炎球菌进行培养。在培养过程中补加葡萄糖溶液、优化搅拌速度及通气量均有利于两菌群生长;在多糖纯化过程中,通过对比试验选择25%乙醇沉淀核酸,2～8℃的酚溶液,萃取蛋白的次数控制在3～4次为好。结论所研制的综合培养基及多糖纯化的工艺具有较好的可行性,为四价脑膜炎球菌多糖疫苗的研制提供了试验基础。

Culture of Meningococcus Serogroups W135 and Y and Purification of Polysaccharide

Objective To culture meningococcus serogroups W_~135 and Y and purify capsular polysaccharide.Methods Meningococcus serogroups W_~135 and Y were cultured in a 75 L bioreactor,and the formula of medium as well as procedures of fermentation and extraction and purification of polysaccharide were studied.Results Synthetic medium might be used for the culture instead of semi-synthetic medium.Supplying glucose solution and optimizing the stirring rate and airing quantity were beneficial to the growth of meningococcus serogroups W_~135 and Y.During the purification of polysaccharide,25% alcohol might be selected to precipitate the nucleic acid.The times of extraction of protein with phenol solution at 2-8℃ should be controlled at 3-4.Conclusion The developed synthetic medium and purification procedure were feasible and provided an experimental basis for the preparation of tetravalent meningococcus polysaccharide vaccine.