PURIFICATION OF OLIGOSACCHARIDE ANTIGENS BY WEAK AFFINITY-CHROMATOGRAPHY

Weak affinity (K(a) almost-equal-to 10(2)-10(4) M-1) monoclonal antibodies immobilized at high concentrations (> 50 mg/mL) on macroporous silica matrices provide a chromatographic medium for high performance separation of oligosaccharide antigens. Unlike classical affinity chromatography, weak affinity chromatography utilizes dynamic equilibrium kinetics as a basis for separation of antigens. Simple calculations based upon established principles of chromatographic theory enable rapid determination of affinity constants and systematic adjustment of column parameters to optimize analytical and semipreparative chromatography. Demonstrated applications of weak affinity chromatography include analysis of picomol quantities of oligosaccharides in complex biological fluids such as urine and serum and recovery of oligosaccharide antigens released from partially fractionated oligosaccharides of pooled human milk.