Purification of Shiga toxin by α-d-galactose-(1 → 4)-β-d-galactose-(1 → 4)- β-d-glucose-(1 →) receptor ligand-based chromatography

Abstract

A simple and rapid method for Shiga toxin purification based on specific binding to the Galα1 → 4Galβ1 → 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 otained, 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. © 1989.