Abundant plasma protein depletion using ammonium sulfate precipitation and Protein A affinity chromatography

Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
Lentel PringelsLiliane Schoofs

Abstract

Plasma is a highly valuable resource for biomarker research since it is easy obtainable and contains a high amount of information on patient health status. Although advancements in the field of proteomics enabled analysis of the plasma proteome, identification of low abundant proteins remains challenging due to high complexity and large dynamic range. In order to reduce the dynamic range of protein concentrations, a tandem depletion technique consisting of ammonium sulfate precipitation and Protein A affinity chromatography was developed. Using this method, 50% of albumin, together with other high abundant proteins such as alpha-1-antitrypsin, was depleted from the plasma sample at 20% to 40% ammonium sulfate saturation levels. In combination with immunoglobulin removal using a Protein A column, this technique delivered up to 40 new low- to medium abundance protein identifications when performing a shotgun mass spectrometry analysis. Compared to non-depleted plasma, 270 additional protein spots were observed during 2D-PAGE analysis. These results illustrate that this tandem depletion method is equivalent to commercial kits which are based on immune-affinity chromatography. Moreover, this method using Protein A immunoglobulin de...Continue Reading

Citations

Sep 18, 2020·Advances in Colloid and Interface Science·Shixiang LiuHailin Cong
Jul 1, 2020·Journal of Proteomics·Corina Itze-Mayrhofer, Gottfried Brem
Aug 8, 2021·International Journal of Molecular Sciences·Katie DunphyDespina Bazou

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