Rapid protein A assay for intrinsic factor and its binding antibody

Abstract

A simple and rapid method for the measurement of cobalamin bound intrinsic factor (Cbl-IF) complex and intrinsic factor binding antibody is described. The method is based on the principle of affinity chromatography and adapted to a batch separation technique. A specific ligand staphylococcal protein A was coupled to Sepharose to form a convenient solid phase matrix. The intrinsic factor binding antibody in patients with pernicious anaemia was used to form an immune complex with Cbl-IF. This complex was adsorbed on to staphylococcal protein A. Gastric juice from control subjects and patients with pernicious anaemia was assayed for intrinsic factor activity and the results correlated very closely with two other established methods. Sera from 30 control subjects were assayed for binding intrinsic factor antibody and all were found to be negative; of 15 patients with pernicious anaemia, six were positive. These patients were selected with blocking antibody. The method does not require technologically advanced protein separation techniques and could therefore be applied in any clinical laboratory using radioisotopes. It could also be adapted to assay cobalamin in body fluids or in food.