Lewis A C, Freeman B A
Science. 1969 Aug 22;165(3895):808-9. doi: 10.1126/science.165.3895.808.
Choleragenic toxin was separated from vascular permeability factor by ion-exchange chromatography of supernatants of dialyzed peptone cultures of Vibrio cholerae. The choleragenic toxin eluted from columns of QAE-Sephadex with low-ionicity systems is free of permeability factor activity. Further elution of these columns with 0.5M NaCl removes both the permeability factor and residual choleragenic toxin. When this latter material is chromatographed on columns of carboxymethyl-Sephadex, the permeability factor toxin is eluted by 0.02M phosphate buffer and is free of choleragenic activity. Therefore, choleragenic and permeability factor activities of the type 2 cholera toxins are different and can be separated by these procedures.
通过对霍乱弧菌透析蛋白胨培养上清液进行离子交换色谱法,从血管通透性因子中分离出霍乱毒素。用低离子强度系统从QAE - 葡聚糖凝胶柱上洗脱下来的霍乱毒素没有通透性因子活性。用0.5M氯化钠进一步洗脱这些柱子,可去除通透性因子和残留的霍乱毒素。当将后一种物质在羧甲基葡聚糖凝胶柱上进行色谱分析时,通透性因子毒素被0.02M磷酸盐缓冲液洗脱,且没有霍乱毒素活性。因此,2型霍乱毒素的霍乱毒素活性和通透性因子活性是不同的,并且可以通过这些方法分离。