Lewis A C, Richardson S H, Sheridan B
Appl Environ Microbiol. 1976 Aug;32(2):288-93. doi: 10.1128/aem.32.2.288-293.1976.
Procedures for cholera enterotoxin purification previously developed in this labarotory were not applicable to large-scale purification, and these methods resulted in low yields of pure toxin. An efficient scheme has been developed whereby pure cholera enterotoxin can be obtained from 6 to 8 liters of culture supernatant fluid. This method consists of concentration by membrane ultrafiltration followed by gel filtration and cation-exchange chromatography. Pure cholera enterotoxin of high biological potency was obtained after a final step of preparative acrylamide gel electrophoresis. The degree of purity of the toxin-antigen as well as its biological activity were determined at various setps of purification. This alternate technique for purification is offered because of the widespread interest in cholera enterotoxin as a specific stimulator of adenyl cyclase.
本实验室先前开发的霍乱肠毒素纯化程序不适用于大规模纯化，且这些方法导致纯毒素产量较低。现已开发出一种高效方案，可从6至8升培养上清液中获得纯霍乱肠毒素。该方法包括通过膜超滤浓缩，然后进行凝胶过滤和阳离子交换色谱。在制备性丙烯酰胺凝胶电泳的最后一步之后，获得了具有高生物活性的纯霍乱肠毒素。在纯化的各个步骤中测定了毒素抗原的纯度及其生物活性。由于人们对霍乱肠毒素作为腺苷酸环化酶的特异性刺激剂广泛感兴趣，因此提供了这种替代纯化技术。
