Multi-dimensional liquid chromatography in proteomics—A review

Abstract

Proteomics is the large-scale study of proteins, particularly their expression, structures and functions. This still-emerging combination of technologies aims to describe and characterize all expressed proteins in a biological system. Because of upper limits on mass detection of mass spectrometers, proteins are usually digested into peptides and the peptides are then separated, identified and quantified from this complex enzymatic digest. The problem in digesting proteins first and then analyzing the peptide cleavage fragments by mass spectrometry is that huge numbers of peptides are generated that overwhelm direct mass spectral analyses. The objective in the liquid chromatography approach to proteomics is to fractionate peptide mixtures to enable and maximize identification and quantification of the component peptides by mass spectrometry. This review will focus on existing multidimensional liquid chromatographic (MDLC) platforms developed for proteomics and their application in combination with other techniques such as stable isotope labeling. We also provide some perspectives on likely future developments.

The Combination of Lectin Affinity Chromatography, Gel Electrophoresis and Mass Spectrometry in the Study of Plant Glycoproteome: Preliminary Insights

“…Various techniques (liquid chromatography, capillary, gel electrophoresis, or different varieties of affinity chromatography) have been widely used to enrich and purify the proteins of interest from other proteins and components (e.g., [7][8][9][10][11][12]). The most powerful of these methods is affinity chromatography (AC).…”

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“…The most widely used high throughput separation technique is the coupling of IEX and RPC with MS instrumentation. Other examples of multidimensional HPLC/MS techniques can also include RPC/CE chromatography, affinity chromatography/RP chromatography, SEC/IEX/RPC and SEC/ RPC/CE (Issaq et al, 2005;Zhang et al, 2010a).…”

Proteomic approaches to the study of renal mitochondria

“…Proteolytic digestion of complex samples results in huge number of proteolytic peptides and multidimensional fractionation of peptides based on combination of two or more chromatographic steps (e.g., reverse phase liquid chromatography, ion exchange chromatography) (ref. 55 ) offers increased separation capacity and higher resolution. Separated peptides are eluted from the column directly into the mass spectrometer.…”