Pharmaceutical Sciences Faculty Publications

Title

Methods for Fractionation, Separation and Profiling of Proteins and Peptides

Document Type

Article

Publication Date

9-1-2002

Journal Title

Electrophoresis

ISSN

0173-0835

Volume

23

Issue

17

First Page

3048

Last Page

3061

DOI

10.1002/1522-2683(200209)23:17<3048::AID-ELPS3048>3.0.CO;2-L

PubMed ID

12207315

Abstract

In the last few years there has been an increased effort to develop technologies capable of identifying and quantifying large numbers of proteins expressed within a cell system (i.e., the proteome). The complexity of the mixtures being analyzed has made the development of effective fractionation and separation methods a critical component of this effort. This review highlights many of the protein and peptide fractionation and separation methods, such as electrophoresis and high-performance liquid chromatography (HPLC), which have experienced significant development over the past forty years. Modern instrumental strategies for the resolution of cell proteins, based on separations employing a single high-resolution or multidimensional approach, and the relative merits of each, will be discussed. The focus of this manuscript will be on the development of multidimensional separations such as two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), HPLC/HPLC, and HPLC-capillary electrophoresis and their application to the characterization of complex proteome mixtures.

Keywords

Chromatography, electrophoresis, peptides, proteins

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