Protein/peptide purification by three-well OFFGEL electrophoresis with immobilized ultra narrow pH gradient gels

Purification and desalting of protein and peptide samples by three-well OFFGEL electrophoresis with immobilized ultra narrow pH gradient gels is proposed as a fast preparative strategy for proteomics. The gist of this strategy is to separate proteins and peptides according to their isoelectric point...

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Bibliographic Details
Published inAnalytical methods Vol. 6; no. 12; pp. 3995 - 4002
Main Authors Tobolkina, Elena, Cortés-Salazar, Fernando, Qiao, Liang, Girault, Hubert H.
Format Journal Article
LanguageEnglish
Published 01.01.2014
Subjects
Electrophoresis
Gels
Mathematical analysis
Peptides
Proteins
Purification
Strategy
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Summary:Purification and desalting of protein and peptide samples by three-well OFFGEL electrophoresis with immobilized ultra narrow pH gradient gels is proposed as a fast preparative strategy for proteomics. The gist of this strategy is to separate proteins and peptides according to their isoelectric points and to isolate those of a given pI value equal to the mean pH value of the gel. The present approach has been demonstrated both on protein mixtures and a digested Escherichia coli protein extract. UV-Vis spectroscopy, MALDI-MS, SDS-PAGE and LC-MS/MS were employed for the quantitative and qualitative characterization of the separation results. The electrophoretic methodology has been simulated by finite element methods.
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ISSN:1759-9660
1759-9679
DOI:10.1039/C4AY00088A