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PubMed · 15309736

Simplified purification method for Clostridium difficile toxin A.

Si-Wu Fu·Jing Xue·Ya-Li Zhang·Dian-Yuan Zhou
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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 pH 5.5, 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 550,000. The purified toxin A had a protein concentration of 0.881 mg/mL. The minimum lethal dose was 1X10(6) MLD/mL (i.p.mice). The cytotoxic titer was 10(7) CU/mg. The haemagglutinate activity was at a concentration of 1.72 microg/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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AnimalsAnion Exchange ResinsBacterial ToxinsBiotechnologyChemical PrecipitationChlorocebus aethiopsChromatography, AffinityEnterotoxinsEthanolaminesIleumImmunodiffusionPolymersRabbitsVero CellsJournal Article

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Si-Wu Fu, Jing Xue, Ya-Li Zhang, Dian-Yuan Zhou. 2004-09-15. Simplified purification method for Clostridium difficile toxin A.. https://doi.org/10.3748/wjg.v10.i18.2756

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