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Copyright ©The Author(s) 2004. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Sep 15, 2004; 10(18): 2756-2758
Published online Sep 15, 2004. doi: 10.3748/wjg.v10.i18.2756
Simplified purification method for Clostridium difficile toxin A
Si-Wu Fu, Jing Xue, Ya-Li Zhang, Dian-Yuan Zhou
Si-Wu Fu, Jing Xue, Ya-Li Zhang, Dian-Yuan Zhou, PLA Institute for Digestive Medicine, Nanfang Hospital, First Military Medical University, Guangzhou 510515, Guangdong Province, China
Author contributions: All authors contributed equally to the work.
Correspondence to: Dr. Si-Wu Fu, PLA Institute for Digestive Medicine, Nanfang Hospital, First Military Medical University, Guangzhou 510515, Guangdong Province, China. fusiwu2001@sina.com
Telephone: +86-20-61641544
Received: December 17, 2003
Revised: December 23, 2003
Accepted: January 12, 2004
Published online: September 15, 2004
Abstract

AIM: To establish the purification method for Clostridium difficile (C. difficile) toxin A.

METHODS: C. difficile VPI 10463 filtrate was cultured anaerobically by the dialysis bag methods. And then the toxin A was purified by precipitation with 500 g/L (NH4)2SO4 and acid precipitation at pH5.5, followed by ion-exchange chromatography on DEAE-Toyopearl.

RESULTS: Purified toxin A exhibited only one band on native polyacrylamide gel electrophoresis (native-PAGE) and Ouchterlony double immunodiffusion. The molecular weight of toxin A was estimated to be 550000. The purified toxin A had a protein concentration of 0.881 mg/mL. The minimum lethal dose was 1 × 106 MLD/mL (i.p.mice). The cytotoxic titer was 107 CU/mg. The haemagglutinate activity was at a concentration of 1.72 μg/mL. The ratio of fluid volume (mL) accumulated to the length (cm) of the loop was 2.46.

CONCLUSION: The modified method for purification of toxin A of C. difficile was simple and convenient. It may be even more suitable for purification of toxin A on large scales.

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