Combining isoelectric point-based fractionation, liquid chromatography and mass spectrometry to improve peptide detection and protein identification.

ABSTRACT: The off-line coupling of an isoelectric trapping device termed membrane separated wells for isoelectric focusing and trapping (MSWIFT) to mass spectrometry-based proteomic studies is described. The MSWIFT is a high capacity, high-throughput, mass spectrometry-compatible isoelectric trapping device that provides isoelectric point (pI)-based separations of complex mixtures of peptides. In MSWIFT, separation and analyte trapping are achieved by migrating the peptide ions through membranes having fixed pH values until the peptide pI is bracketed by the pH values of adjacent membranes. The pH values of the membranes can be tuned, thus affording a high degree of experimental flexibility. Specific advantages of using MSWIFT for sample prefractionation include: (1) small sample volumes (approximat

SUBMITTER: Cologna SM 

PROVIDER: S-EPMC2927729 | biostudies-literature | 2010 Sep

REPOSITORIES: biostudies-literature