Low affinity pair size exclusion chromatography

Low affinity pair size exclusion chromatog. (LAPSEC) is herein described as a novel approach specifically for lectin based monosaccharide sepn. but with the potential for wider application using weak antibody-antigen interactions. This technique exploits weak ligand receptor interactions and the difference in mol. size between free and receptor-bound ligand, to effect sepns. using a size exclusion chromatog. column. While such carrier-based sepns. are achievable with high affinity interactions, the advantage of the approach described here is that the use of weak interactions also allows sepn. of ligand from carrier in the same column, allowing recycling of the carrier. The utility of the LAPSEC approach is shown by results obtained using Con A and Lotus Tetragonolobus lectin to sep. their specific monosaccharides (D-mannose and L-fucose, resp.) from unbound monosaccharides. These systems have been simulated using a simple multi-sectional equil. model using independent measurements of binding consts. The results show that monosaccharide sepn. and recovery of lectin is feasible in a single run and suggests that this approach may have potential for the selective recovery of low mol. wt. products on a preparative scale. As receptors are not immobilized, resin costs are reduced, there is efficient use of affinity receptor, and sepn. is achieved under isocratic conditions. [on SciFinder (R)]