艰难梭菌A毒素的纯化

Purification of Clostridium difficile toxin A

目的建立一种纯化艰难梭菌A毒素的方法。方法将艰难梭菌VPI 10 4 6 3菌株经透析培养 ,5 0 %饱和硫酸胺盐析 ,酸沉淀 ,再经DEAE Toyopearl 6 5 0M离子交换色谱。结果精制的艰难梭菌A毒素Native PAGE及对流免疫电泳均为单一带 ,相对分子质量为 5 5 0 0 0 0 ,小鼠毒力为 1× 10 6MLD/ml,Vero细胞毒性为 1× 10 7CU/ml。凝血活性为 1∶5 12 ,肠袢试验为阳性。结论该法简便、实用、可行。

PurposeTo establish the purification method of Clostridium difficile toxin A.MethodsThe purification process included precipitation with 50% (NH 4) 2SO 4 and acetic acid precipitation at pH 5.5, followed by ion-exchange chromatography on DEAE-Toyopearl 650M.ResultsThe purified toxin A exhibited only one band on native polyacrylamide gel electrophoresis(Native-PAGE) and counterimmunelectrophoresis,and the molecular weight of toxin A was estimated to be 550 000.ConclusionHighly purified toxin A of Clostridium difficile was obtained by a simple modified method.