Remarkable effect of mobile phase buffer on the SEC-ICP-AES derived Cu, Fe and Zn-metalloproteome pattern of rabbit blood plasmaElectronic supplementary information (ESI) available: Table S1 - Retention time and peak area data for the Cu, Fe and Zn peaks of all mobile phases. See DOI: 10.1039/c003321aThroughout this manuscript a metalloprotein is defined as a protein which contains either a weakly or strongly bound metal irrespective of the biochemical role that the metalloprotein serves in the

• Main Authors: Jahromi, Elham Zeini, White, Wade, Wu, Qiao, Yamdagni, Raghav, Gailer, Jürgen
• Format: Article
• Language: English
• ISSN: 1756-5901; 1756-591X
• DOI: 10.1039/c003321a

The development of an analytical method to quantify the major Cu, Fe and Zn-containing metalloproteins in mammalian plasma has been recently reported. This method is based on the separation of plasma proteins by size exclusion chromatography (SEC) followed by the on-line detection of the metalloproteins by an inductively coupled plasma atomic emission spectrometer (ICP-AES). To assess whether the mobile phase buffer can affect the SEC-ICP-AES-derived metalloproteome pattern, thawed rabbit plasma was analyzed using phosphate buffered saline (PBS)-buffer (0.15 M, pH 7.4), Tris-buffer (0.1 and 0.05 M, pH 7.4), Hepes-buffer (0.1 M, pH 7.4) or Mops-buffer (0.1 M, pH 7.4). In contrast to the Cu-specific chromatograms, the Fe and Zn-specific chromatograms that were obtained with Tris, Hepes and Mops-buffer were considerably different from those attained with PBS-buffer. The Tris, Hepes and Mops-buffer mediated redistribution of ∼25% plasma Zn 2+ from 100-600 kDa plasma proteins and to a smaller extent to a