Purification of human intrinsic factor using high-performance ion-exchange chromatography as the final step.
Human intrinsic factor was purified 1430-fold from gastric juice with a yield of 75% using two steps: labile ligand affinity chromatography and high-performance ion-exchange chromatography. Intrinsic factor precipitated in the presence of specific autoantibodies and 15% sodium sulfate, had an estimated Mr of 59 000 in 5% SDS electrophoresis and could bind to the specific ileal receptor in vitro. Its carbohydrate composition could be related to N-lactosaminic and O-glycosidic chains. High-performance ion-exchange chromatography was a mild, rapid and efficient procedure to separate completely intrinsic factor from haptocorrin (another glycoprotein of gastric juice which binds cobalamin) and from other contaminating proteins.
D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D005750:Gastric Juice / Q000032:analysis; D006801:Humans; D007437:Intrinsic Factor / Q000032:analysis / Q000302:isolation & purification*
D007437:Intrinsic Factor