Simple method for Shiga toxin 2e purification by affinity chromatography via binding to the divinyl sulfone group

PloS One
Hideyuki ArimitsuTakao Tsuji

Abstract

Here we describe a simple affinity purification method for Shiga toxin 2e (Stx2e), a major causative factor of edema disease in swine. Escherichia coli strain MV1184 transformed with the expression plasmid pBSK-Stx2e produced Stx2e when cultivated in CAYE broth containing lincomycin. Stx2e bound to commercial D-galactose gel, containing α-D-galactose immobilized on agarose resin via a divinyl sulfone linker, and was eluted with phosphate-buffered saline containing 4.5 M MgCl2. A small amount of Stx2e bound to another commercial α-galactose-immobilized agarose resin, but not to β-galactose-immobilized resin. In addition, Stx2e bound to thiophilic adsorbent resin containing β-mercaptoethanol immobilized on agarose resin via a divinyl sulfone, and was purified in the same manner as from D-galactose gel, but the Stx2e sample contained some contamination. These results indicate that Stx2e bound to D-galactose gel mainly through the divinyl sulfone group on the resin and to a lesser extent through α-D-galactose. With these methods, the yields of Stx2e and attenuated mutant Stx2e (mStx2e) from 1 L of culture were approximately 36 mg and 27.7 mg, respectively, and the binding capacity of the D-galactose gel and thiophilic adsorbent res...Continue Reading

References

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Nov 20, 2012·Microbiology and Immunology·Hideyuki ArimitsuTakao Tsuji

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Citations

Feb 24, 2016·Toxicon : Official Journal of the International Society on Toxinology·Robert Alvin Bernedo-Navarro, Tomomasa Yano

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Datasets Mentioned

BETA
GU459254

Methods Mentioned

BETA
protein assay
enzyme-linked immunosorbent assay
ELISA
electrophoresis
affinity purification

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