Methods for fractionation, separation and profiling of proteins and peptides

Haleem J. Issaq*, Thomas P. Conrads, George M. Janini, Timothy D. Veenstra

*Corresponding author for this work

Research output: Contribution to journalReview articlepeer-review

190 Scopus citations


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 mixture.

Original languageEnglish
Pages (from-to)3048-3061
Number of pages14
Issue number17
StatePublished - Sep 2002
Externally publishedYes


  • Fractionation
  • Multidimensional separations
  • Peptides
  • Proteins
  • Proteomics
  • Review


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