title: Application of liquid-liquid partition chromatography in the simultaneous purification of sex-hormone-binding globulin and corticosteroid-binding globulin.
abstract: Two human serum proteins, corticosteroid-binding globulin (CBG) and sex-hormone-binding globulin (SHBG), were purified to homogeneity by the application of a combination of three different modes of chromatography. Human pregnancy serum was fractionated with ammonium sulphate. SHBG (50% pellet) and CBG (80% pellet) were then purified by affinity chromatography on tresyl-activated Sepharose with 15-aminopentadecanoic acid (for SHBG) and 1,12-diaminododecane (for CBG) as spacers and 17 zeta-aminoethyl-5 alpha-androstan-3 beta,17-diol (for SHBG) and 17 alpha-hydroxy-4-androsten-3-one-17 beta-carboxylic acid (for CBG) as specific ligands for these two proteins. The eluate was injected into a Mono Q anion-exchange column. Fractions containing SHBG or CBG were finally purified by liquid-liquid chromatography on Lipar-Gel 750. This chromatographic sequence clearly separated SHBG and CBG from other proteins, mainly serum albumin, without a loss of protein or binding activity.
authors.affiliation: Abteilung für Experimentelle Endokrinologie, Johannes Gutenberg Universität Mainz, F.R.G.
authors.forename: A
authors.initials: A
authors.lastname: Heubner
authors.forename: O
authors.initials: O
authors.lastname: Belovsky
authors.forename: W
authors.initials: W
authors.lastname: Müller
authors.forename: H J
authors.initials: HJ
authors.lastname: Grill
authors.forename: B
authors.initials: B
authors.lastname: Manz
authors.forename: M
authors.initials: M
authors.lastname: Juchem
authors.forename: K
authors.initials: K
authors.lastname: Pollow
mesh_terms: D002846:Chromatography, Affinity; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D007525:Isoelectric Focusing; D011247:Pregnancy; D011956:Receptors, Cell Surface / Q000032:analysis; D012738:Sex Hormone-Binding Globulin / Q000302:isolation & purification*; D013056:Spectrophotometry, Ultraviolet; D014156:Transcortin / Q000302:isolation & purification*
chemical_list: D011956:Receptors, Cell Surface; D012738:Sex Hormone-Binding Globulin; D014156:Transcortin