High-yield high-performance liquid chromatographic analysis of steroid hormone receptors on glass columns

Abstract

The HPLC characteristics of extensively purified chicken oviduct protesterone receptors were compared on TSK 3000 SW size-exclusion and DEAE-5-PW media packed in either glass or stainless-steel columns. Recoveries of [ 3H]progesterone-labeled receptor from size exclusion were 75–95% in glass columns and less than 10% in stainless-steel columns. Similarly, recoveries from DEAE were greater than 90% in glass columns but only ∼45% in stainless-steel columns. Recoveries in glass columns were similar on several HPLC systems. Thus, the requisite component for high yields from extensively purified receptor preparations was the glass column itself. While receptor B exhibited ionic strength-dependent mobility similar to several standard proteins on size-exclusion glass column HPLC, receptor A was very peculiar. Resolution of receptors A and B was superior to previous reports using size exclusion open-end chromatography. We also resolved functionally active proteolytic gragments. Finally, the generality of glass column size-exclusion HPLC was demonstrated by high-yield analyses of different steroid hormone receptors from different tissues and species.