Home General Physiology and Biophysics 2019 General Physiology and Biophysics Vol.38, No.4, p.305–314, 2019

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Quarterly, 80 pp. per issue
Founded: 1982
ISSN  1338-4325 (online)

Published in English

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General Physiology and Biophysics Vol.38, No.4, p.305–314, 2019

Title: A comparison of albumin removal procedures for proteomic analysis of blood plasma
Author: Anna Tomascova, Jan Lehotsky, Dagmar Kalenska, Eva Baranovicova, Peter Kaplan, Zuzana Tatarkova

Abstract: Blood biomarkers are usually present in low concentration and can be masked by the high-abundance proteins, of which albumin is the predominant one. The purpose of this study was to compare four different albumin removal methods compatible with in-gel based proteomics, applicable for plasma, without requiring specific techniques and high financial input. Plasma underwent albumin depletion with ultrafiltration device Amicon Ultra, commercial ProteoPrep Blue Albumin and IgG Depletion Kit, acetonitrile precipitation method and precipitation with acetonitrile-methanol protocol. All samples were evaluated by 1-D and 2-D gel electrophoresis with subsequent mass spectrometry protein identification. Two of the tested methods (ProteoPrep BlueKit and acetonitrile-methanol precipitation) maintained sufficient protein content for further in-gel analyses. Their 2-D protein profiles were distinctively separated and overlapped with protein profile of crude plasma. Protein spot count showed significant increase in protein spots, compared to crude plasma, only with acetonitrile-methanol precipitation method. Precipitation with acetonitrile-methanol method significantly increased number of protein spots on 2-D protein profile and improved score of mass spectrometry identification. However, albumin was still present and found in number of protein spots.

Keywords: Albumin removal — Mass spectrometry — Biomarkers — Blood plasma
Published online: 15-Jul-2019
Year: 2019, Volume: 38, Issue: 4 Page From: 305, Page To: 314

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