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A molecular scanner to automate proteomic research and to display proteome images

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Published in Analytical chemistry. 1999, vol. 71, no. 21, p. 4981-8
Abstract Identification and characterization of all proteins expressed by a genome in biological samples represent major challenges in proteomics. Today's commonly used high-throughput approaches combine two-dimensional electrophoresis (2-DE) with peptide mass fingerprinting (PMF) analysis. Although automation is often possible, a number of limitations still adversely affect the rate of protein identification and annotation in 2-DE databases: the sequential excision process of pieces of gel containing protein; the enzymatic digestion step; the interpretation of mass spectra (reliability of identifications); and the manual updating of 2-DE databases. We present a highly automated method that generates a fully annoated 2-DE map. Using a parallel process, all proteins of a 2-DE are first simultaneously digested proteolytically and electro-transferred onto a poly(vinylidene difluoride) membrane. The membrane is then directly scanned by MALDI-TOF MS. After automated protein identification from the obtained peptide mass fingerprints using PeptIdent software (http://www.expasy.ch/tools/peptident.html + ++), a fully annotated 2-D map is created on-line. It is a multidimensional representation of a proteome that contains interpreted PMF data in addition to protein identification results. This "MS-imaging" method represents a major step toward the development of a clinical molecular scanner.
Keywords AutomationBlood Proteins/analysis/chemistryElectrophoresis, Gel, Two-Dimensional/methodsElectrophoresis, Polyacrylamide Gel/methodsHumansImage Processing, Computer-Assisted/instrumentation/methodsPeptide Mapping/methodsSpectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methodsTrypsin Inhibitor, Kunitz Soybean/analysis/chemistry
PMID: 10565287
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Article (Published version) (311 Kb) - public document Free access
Research groups Calipho (80)
Chimie et protéomique clinique (270)
Groupe de Protéomique biomédicale (635)
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BINZ, Pierre-Alain et al. A molecular scanner to automate proteomic research and to display proteome images. In: Analytical chemistry, 1999, vol. 71, n° 21, p. 4981-8. doi: 10.1021/ac990449e https://archive-ouverte.unige.ch/unige:80769

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Deposited on : 2016-02-22

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