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Published on: June 2, 2023
A novel and one-step purification of human ceruloplasmin by acharan sulfate affinity chromatography
Youmie Park1, In Sun Lee, Eun Ji Joo
1Natural Products Research Institute, Seoul National University, Seoul, 151-742, Korea.
Abstract:
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.
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