Purification of Shiga toxin by alpha-D-galactose-(1----4)-beta-D-galactose-(1----4)-beta-D-glucose-(1- ---) receptor ligand-based chromatography.
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.
Ryd
M
M
Department of Clinical Bacteriology, Huddinge Hospital, Sweden.
Alfredsson
H
H
Blomberg
L
L
Andersson
A
A
Lindberg
A A
AA
D000818:Animals; D001427:Bacterial Toxins / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002470:Cell Survival / Q000187:drug effects; D002846:Chromatography, Affinity / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D004768:Enterotoxins / Q000302:isolation & purification; D008024:Ligands; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009844:Oligosaccharides; D011956:Receptors, Cell Surface; D011971:Receptors, Immunologic / Q000378:metabolism*; D022603:Shiga Toxins; D012762:Shigella dysenteriae; D014709:Vero Cells
D001427:Bacterial Toxins; D004768:Enterotoxins; D008024:Ligands; D009844:Oligosaccharides; D011956:Receptors, Cell Surface; D011971:Receptors, Immunologic; D022603:Shiga Toxins; C045390:shigella toxin receptor