Separation of Vitamin B12-Binding Proteins of Serum, Gastric Juice and Saliva by Rapid DEAE Cellulose Chromatography

Abstract

A method of DEAE cellulose chromatography is described whereby the main vitamin B₁₂ binders in serum, gastric juice and saliva may be separated into two distinct fractions, one containing alpha-globulin binder and the other containing beta-globulin binder, in less than 2 hours. Typical elution patterns of alpha-globulin and beta-globulin B₁₂ binders from normal (⅘ beta; ⅕ alpha), pernicious anemia (⅔ beta; ⅓ alpha), and chronic myeloid leukemia (CML) (¾ alpha) sera are presented. Chromatography of normal gastric juice produced two peaks of B₁₂-binding material, one containing all the intrinsic factor (IF) B₁₂ binder (as well as some non-IF binder), and the other consisting almost exclusively of non-IF B₁₂ binders. Pernicious anemia gastric juice also produced two peaks, but IF was absent. Saliva had one main peak of B₁₂-binding material which seemed to correspond to the non-IF binder of normal gastric juices; both of these eluted with the alpha-globulin fraction.