Basic Information

Abstract Number: 1770-2     Author Name: Brenna E McJury Affiliation: University of North Carolina Session Title: Young Investigator Award from Subdivision on Chromatography and Separation Chemistry of the Analytical Chemistry Division of the ACS Event Type: Award Event Title: Combined LC-LC-MS for the Separation and Identification of Proteins in Complex Mixtures Presider(s): Bidlingmeyer, Brian A Start Time: 08:45 AM ( Slot # 4 ) Date: Wednesday, March 3rd, 2010 Location: 300 Keywords: Capillary LC, Liquid Chromatography/Mass Spectroscopy, Protein, Proteomics

Co-Authors

Name Affiliation Jorenson, James W University of North Carolina Matthew, Brian University of North Carolina Stobaugh, Jordan University of North Carolina

Abstract Content

Two-dimensional LC-LC is an ideal separation system to be coupled with electrospray ionization mass spectrometry. This is especially true if the second dimension is reversed phase LC as the reversed phase mobile phase can be highly compatible with the electrospray process. We are currently pursuing on-line LC-LC-MS of intact proteins from complex mixtures as a means to identify which proteins in the mixture merit further study. The mass spectrometer provides the means for detection and quantification of intact proteins, as well as providing the molecular weight of the proteins. Parallel to mass spectrometric detection, fractions are collected in order to permit subsequent analysis and identification of any proteins of interest. Protein fractions selected for further study are digested with trypsin, and the tryptic peptides analyzed by reversed phase LC-MS-MS, permitting determination of peptide sequences and positive identification of the proteins of interest. We have recently also begun development of LC-LC-MS systems for separation of intact membrane-bound proteins.