Document Type
Article
Publication Date
1-1-1985
Abstract
Chromatofocusing was used to purify the vitamin D-binding protein (DBP) from pig plasma in a procedure that consisted of an initial DEAE-cellulose chromatography followed by DEAE-Sephadex chromatography, with final purification by chromatofocusing. The protein was purified 184-fold over its concentration in plasma. When the plasma was labelled with a tracer concentration of [3H]calcidiol, it was apparent that holo- and apo-DBP did not co-chromatograph on chromatofocusing. The separation of these two forms of DBP on chromatofocusing was verified by using purified apo-DBP mixed with either a tracer or a saturating concentration of calcidiol. This separation was consistent with differences observed in their isoelectric points. The ability to separate apo and holo forms of DBP should permit the study of their specific interactions with other binding proteins and help determine the physiological relevance of these interactions.
Publication Source (Journal or Book title)
Biochemical Journal
First Page
669
Last Page
674
Recommended Citation
Keenan, M., & Holmes, R. (1985). Chromatofocusing in the purification and separation of apo- and holo-(vitamin D-binding protein). Biochemical Journal, 229 (3), 669-674. https://doi.org/10.1042/bj2290669