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Remarkable effect of mobile phase buffer on the SEC-ICP-AES derived Cu, Fe and Zn-metalloproteome pattern of rabbit blood plasma

Content Provider	Royal Society of Chemistry (RSC)
Author	Yamdagni, Raghav Wu, Qiao White, Wade Gailer, Jürgen Jahromi, Elham Zeini
Copyright Year	2010
Abstract	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 Zn2+ from <100 kDa to >100–600 kDa plasma proteins and to a smaller extent to a <10 kDa (Tris)2Zn2+-complex can be rationalized in terms of the abstraction of Zn2+ from the weak binding site on albumin. In contrast, only Hepes and Mops-buffer redistributed ∼20% of plasma Fe3+ from the <100 kDa to the >600 kDa elution range. Based on these results and considering that the utilization of PBS-buffer has previously resulted in the detection of a number of Cu, Fe and Zn-containing metalloentities in rabbit plasma that was most consistent with literature data, this mobile phase buffer is recommended for metallomic studies regarding mammalian blood plasma.
Starting Page	460
Ending Page	468
Page Count	9
File Format	HTM / HTML PDF
ISSN	17565901
Volume Number	2
Issue Number	7
Journal	Metallomics
DOI	10.1039/c003321a
Language	English
Publisher	Royal Society of Chemistry
Access Restriction	Open
Subject Keyword	PBS Phosphate-buffered saline Hepes Albumin Inductively coupled plasma atomic emission spectroscopy Spectrometer Tris Blood plasma SEC Cu Inductively coupled plasma PH Size-exclusion chromatography U.S. Securities and Exchange Commission
Content Type	Text
Resource Type	Article
Subject	Chemistry Medicine Metals and Alloys Biochemistry Biomaterials Biophysics

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