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Selenoprotein P analysis in human plasma - A discrepancy between HPLC fractionation of human plasma with heparin-affinity chromatography and SDS-PAGE with immunoblot analysis

Citation

Jacobson, GA and Featherstone, AM and Townsend, AT and Lord, RJ and Peterson, GM, Selenoprotein P analysis in human plasma - A discrepancy between HPLC fractionation of human plasma with heparin-affinity chromatography and SDS-PAGE with immunoblot analysis, Biological Trace Element Research, 107, (3) pp. 213-220. ISSN 0163-4984 (2005) [Refereed Article]


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DOI: doi:10.1385/BTER:107:3:213

Abstract

Several recent analytical methods for determination of Se and selenoprotein P have involved high-performance liquid chromatography (HPLC) using heparin-affinity columns coupled to inductively coupled plasma-mass spectrometry (ICP-MS) for Se detection. HPLC-ICP-MS chromatography using tandem HPLC columns with ICP-MS detection was used to detect the major selenium-containing proteins in plasma (glutathione peroxidase, albumin, and selenoprotein P). The efficiency of HPLC separation of plasma selenoprotein P was investigated by analyzing HPLC fractions using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with immunoblot analysis. The HPLC fraction corresponding to selenoprotein P contained 25.1% of total selenoprotein P as measured by immunoblot analysis. The majority (74.9%) of total selenoprotein P found by immunoblot analysis was contained in the early HPLC fractions, consistent with either poor heparin affinity, which was not evident based on the HPLC-ICP-MS technique alone or nonspecific binding of the antibody. Immunoblot analysis of selenoprotein relies on antibodies binding to a selenoprotein P epitope, which might be preserved when selenoprotein P is broken down to release selenocysteine residues. Immunoblot methods overestimate selenoprotein P and are not suitable for determinations of intact selenoprotein P. © Copyright 2005 by Humana Press Inc. All rights of any nature, whatsoever, reserved.

Item Details

Item Type:Refereed Article
Research Division:Medical and Health Sciences
Research Group:Pharmacology and Pharmaceutical Sciences
Research Field:Pharmaceutical Sciences
Objective Division:Health
Objective Group:Other Health
Objective Field:Health not elsewhere classified
Author:Jacobson, GA (Dr Glenn Jacobson)
Author:Featherstone, AM (Dr Alison Featherstone)
Author:Townsend, AT (Associate Professor Ashley Townsend)
Author:Lord, RJ (Dr Roger Lord)
Author:Peterson, GM (Professor Gregory Peterson)
ID Code:38275
Year Published:2005
Web of Science® Times Cited:6
Deposited By:Pharmacy
Deposited On:2005-08-01
Last Modified:2011-11-03
Downloads:3 View Download Statistics

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