Affinity purification of a glucose-containing oligosaccharide using a monoclonal antibody.

Binding of a human urinary tetrasaccharide (Glc alpha 1-6Glc alpha 1-4Glc alpha 1-4Glc) by a mouse monoclonal antibody, 61.1, shows an unusually large dependence upon temperature. Association constants determined by equilibrium dialysis double for each 8 degrees C downward shift in temperature from 37 degrees C (8 X 10(3) M-1) to 4 degrees C (1.7 X 10(5) M-1). Purified 61.1 antibody behaves as a specific temperature-sensitive affinity adsorbent when covalently bound to cyanogen bromide-activated Sepharose or non-covalently bound via its Fc portion to staphylococcal protein A-Sepharose. A column containing 72 mg of antibody 61.1 bound to 5 ml of staphylococcal protein A-Sepharose can bind up to 0.52 mumol of reduced tetrasaccharide. The relatively large combining sites of most antibodies may permit chromatographic selection based upon structural features more complex than those recognized by most plant lectins.