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Purification of Shiga toxin by alpha-D-galactose-(1----4)-beta-D-galactose-(1----4)-beta-D-glucose-(1- ---) receptor
M Ryd1, H Alfredsson, L Blomberg
1Department of Clinical Bacteriology, Huddinge Hospital, Sweden.
FEBS Letters
|December 4, 1989
Abstract:
A simple and rapid method for Shiga toxin purification based on specific binding to the Gal alpha 1----4Gal beta 1----4Glc globotrioside trisaccharide covalently linked to polyacryl/polyvinyl (Fractogel) has been developed. A cell-free sonicate-filtrate of Shigella dysenteriae type 1, strain 114Sd was passed over the globotrioside-Fractogel column, and bound toxin eluted with 6 M guanidine-HCl. A yield of 36 mg pure toxin/1 sonicate-filtrate was obtained, i.e. a one step 1224-fold purification. The recovery of biologically active toxin was 87%. The toxin was purified to homogeneity as shown by native PAGE and SDS-PAGE.