A novel and one-step purification of human ceruloplasmin by acharan sulfate affinity chromatography.
Human ceruloplasmin, a copper binding alpha(2)-glycoprotein, was purified by a single-step procedure using acharan sulfate affinity chromatography. Acharan sulfate was immobilized to amine-functionalized agarose matrix through carboxylic acids. Ceruloplasmin in human plasma was obtained from 0.4 M NaCl salt elution and characterized by SDS-PAGE (132 and 125 kDa), isoelectric focusing (pI 4.6), Western blotting, and MALDI-TOF-MS peptide mass fingerprinting. Ceruloplasmin was purified 106 fold with a specific oxidase activity of 0.53 U/mg protein.
Park
Youmie
Y
Natural Products Research Institute, Seoul National University, Seoul, 151-742, Korea.
Lee
In Sun
IS
Joo
Eun Ji
EJ
Hahn
Bum-Soo
BS
Kim
Yeong Shik
YS
D015153:Blotting, Western; D002570:Ceruloplasmin / Q000737:chemistry / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D006025:Glycosaminoglycans / Q000737:chemistry*; D006801:Humans; D007525:Isoelectric Focusing; D008970:Molecular Weight; D010449:Peptide Mapping; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D006025:Glycosaminoglycans; C106771:acharan sulfate; D002570:Ceruloplasmin