Isolation of thyroxine-binding globulin (TBG) by immunoadsorption chromatography: some physical and immunochemical characteristics of TBG.
Thyroxine-binding globulin (TBG) was isolated from pooled whole human serum by diethylaminoethyl (DEAE) Sephadex anion exchange chromatography followed by immunoadsorption chromatography on a cyanogen bromide-activated Sepharose 4B-sheep anti-human TBG immunoadsorbent. Sodium dodecyl sulphate (SDS)-poly-acrylamide gel electrophoresis (PAGE) of the purified TBG revealed a major protein band with a molecular mass of 65 000 and a weak band of molecular mass 54 000 in both reducing and non-reducing buffers. Sedimentation velocity analysis revealed an S20,w coefficient of 4.5S and a calculated molecular mass of 60 000. Immunochemical analysis confirmed the purity of the TBG preparation which gave a single precipitin peak on two-dimensional immunoelectrophoresis.
D000596:Amino Acids / Q000032:analysis; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D005779:Immunodiffusion; D007122:Immunoelectrophoresis; D007163:Immunosorbent Techniques; D008970:Molecular Weight; D013975:Thyroxine-Binding Proteins / Q000302:isolation & purification*; D014461:Ultracentrifugation
D000596:Amino Acids; D013975:Thyroxine-Binding Proteins