Affinity purification of human plasma vitamin D-binding protein

Abstract

During the course of our studies to probe the vitamin D ligand-binding domains of vitamin D-binding pro-tein and vitamin D receptor, we developed a synthetic procedure to modify the 3β-hydroxyl group of vitamin D3 and its 25-hydroxy- and 1,25-dihydroxy metabolites with a 3-aminopropylether group. In the present study we have coupled 25-hydroxyvitamin D3-3β-3΄-aminopropylether to an activated Sepharose matrix. Using this stable and reusable affinity matrix we have purified human vitamin D-binding protein from human plasma to homogeneity. © Academic Press, Inc.