Use of monolithic supports in proteomics technology

An overview on the utilization of monoliths in proteomics technology will be given. Both silica- and polymer-based monoliths have broad use for microseparation of tryptic peptides in reversed-phase (RP) mode before identification by mass spectrometry (MS) or by MS/MS. For two-dimensional (2D) LC sep...

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Bibliographic Details
Published in:Journal of Chromatography A 2007-03, Vol.1144 (1), p.2-13
Main Authors: Josic, Djuro, Clifton, James G.
Format: Article
Language:English
Subjects:
Analytical, structural and metabolic biochemistry
Biological and medical sciences
Chromatography, Affinity
Chromatography, Liquid - instrumentation
Chromatography, Liquid - methods
Fundamental and applied biological sciences. Psychology
General aspects, investigation methods
Microscopy, Electron, Scanning
Monoliths
Nanoparticles - chemistry
Nanoparticles - ultrastructure
Polymers - chemistry
Protein identification
Proteins
Proteomics
Proteomics - instrumentation
Proteomics - methods
Sample preparation
Silicon Dioxide - chemistry
Tandem Mass Spectrometry - instrumentation
Tandem Mass Spectrometry - methods
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Summary:An overview on the utilization of monoliths in proteomics technology will be given. Both silica- and polymer-based monoliths have broad use for microseparation of tryptic peptides in reversed-phase (RP) mode before identification by mass spectrometry (MS) or by MS/MS. For two-dimensional (2D) LC separation of peptides before MS or MS/MS analysis, a combination of ion-exchange, usually cation-exchange (CEX) chromatography with RP chromatography on monolithic supports can be employed. Immobilized metal ion affinity chromatography monoliths with immobilized Fe 3+-ions are used for the isolation of phosphopeptides. Monoliths with immobilized affinity ligands are usually applied to the rapid separation of proteins and peptides. Miniaturized reactors with immobilized proteolytic enzymes are utilized for rapid on- or offline digestion of isolated proteins or protein mixtures prior to identification by LC–MS/MS. Monoliths also have broad potential for application in sample preparation, prior to further proteomic analyses. Monolithic supports with large pore sizes can be exploited for the isolation of nanoparticles, such as cells, organelles, viruses and protein aggregates. The potential for further adoption of monolithic supports in protein separation and enrichment of low abundance proteins prior to proteolytic digestion and final LC–MS/MS protein identification will be discussed.
ISSN:0021-9673
DOI:10.1016/j.chroma.2006.11.082