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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。
Objective To cultivate the purification of meningococci and capsular polysaccharides of W135 and Y groups. Methods Two groups of meningococci were cultured in a 75 L bioreactor using a synthetic medium instead of a semi-synthetic medium to study medium preparation, fermentation, polysaccharide extraction and purification. Results The integrated medium could replace two groups of meningococci in semi-synthetic medium. In the course of cultivation, adding glucose solution, optimizing stirring speed and aeration volume are conducive to the growth of the two bacteria; in the process of polysaccharide purification, 25% ethanol was used to precipitate nucleic acid, 2 ~ 8 ℃ phenol solution and protein extraction The number of control in 3 to 4 times as well. CONCLUSION The synthetic medium and polysaccharide purification technology developed are feasible and provide experimental basis for the development of tetravalent meningococcal polysaccharide vaccine.