Beckman
Coulter Application (system and method support)
Proteome Analysis of Human Plasma with the ProteomeLab™ PF 2D System
Michael H. Simonian, Ph.D., and Edna Betgovargez
Beckman Coulter, Inc.
Proteomics is the study of how information coded in a cell is expressed and regulated at the protein level to achieve the function of an organism. Among the goals of proteomics are the following: the identification of all the proteins in a given cell, tissue or organism; determination of how these proteins interact; and, finally, to understand the mechanism of the function of these proteins. An ultimate result of proteomics is the understanding of complex biological systems, which can lead to new diagnostics and therapy. The first step toward this end is the identification of all proteins in a given system by protein mapping or profiling. Typically, this is considered the discovery phase of proteomics and involves the comparison of different states of a cell or tissue, such as diseased vs. normal, mature vs. immature, or drug-tested vs. non-treated. Traditionally, this profiling has been accomplished by two-dimensional gel electrophoresis for the discovery phase of proteomics. Although two-dimensional polyacrylamide gel electrophoresis has successfully resolved proteomics, it is labor-intensive, time-consuming, and at best, semi-quantitative.
This paper describes an alternative approach to two-dimensional gel electrophoresis for the discovery phase of proteomics. A complete system, the ProteomeLab PF 2D has been developed with integrated hardware, software, and chemistry. This system uses two-dimensional liquid chromatography. The first dimension is chromatofocusing, which separates based on the isoelectric points (pI) of proteins. Fractions are collected in the first dimension and, subsequently, are separated by a second dimension of reversed-phase chromatography, which separates based on hydrophobicity. This two-dimensional approach was used to compare the proteins in fasting and non-fasting human plasma and then determine if there were any qualitative and/or quantitative differences between the two states of plasma.
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