Screening, purification, and identification of a copper-dependent FITC-binding protein in human plasma: Albumin

Yu Wei Wu, Sung Fang Chen, Charng Bin Yang, Yu Hui Tsai

研究成果: 雜誌貢獻期刊論文同行評審

6 引文 斯高帕斯(Scopus)

摘要

In this study, a protein purified by fluorescein isothiocyanate (FITC)-affinity chromatography from human plasma was identified as albumin by MALDI-TOF-MS. Albumin was found to conjugate with FITC-labeled molecules through a copper-dependent reaction. The formation of this complex was confirmed by methods including a newly developed "charcoal-based fluorescence assay" (CFA), gel-filtration, affinity chromatography, and ultrafiltration. The binding was identified as disulfide bridge formation. This is the first to demonstrate that copper induces a covalent binding of FITC-labeled molecules with albumin. In addition, the developed CFA method facilitates the screening of small fluorescent dyes binding to macromolecules.

原文英語
頁(從 - 到)187-191
頁數5
期刊Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
863
發行號1
DOIs
出版狀態已發佈 - 2008 二月 15

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry
  • Cell Biology

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