Universal method for the facile production of glycolipid/lipid matrices for the affinity purification of binding ligands

Abstract

Glycolipid recognition is a common motif in cellular physiology and bacterial pathogenesis. Such protein/lipid interactions are most conveniently demonstrated by the thin-layer chromatogram overlay. We have designed a simple affinity matrix for the purification of such glycolipid (or lipid) binding ligands based on the same principle, i.e., glycolipid (or lipid) adsorbed onto silica. The versatility of the procedure is demonstrated by the purification of several anti-glycolipid antibodies and anti-phosphatidyl ethanolamine (anti-PE) and the affinity purification of the Escherichia coli-derived verotoxin which binds to globotriaosyl ceramide.