A displacement chromatography based proteomic analysis platform

ANYL 28

Steven Taylor Evans, evanss3@rpi.edu1, Alexander S. Freed, freeda@rpi.edu1, Mark Platt, plattm@rpi.edu2, and Steven M. Cramer, crames@rpi.edu1. (1) Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, 110 8th St, Troy, NY 12180, (2) Department of Chemistry and Chemical Biology, Rensselaer Polytechnic Institute, Troy, NY
Ion exchange displacement chromatography was employed at both the protein and peptide levels in concert with mass spectrometry to develop a new proteomics analysis platform. Studies were first carried out using model mixtures of proteins and peptides to determine the limits of this approach. These results confirmed that trace solutes could be significantly concentrated and enriched during the displacement process. Further, simulations carried out using a Steric Mass Action based chromatographic model were in good agreement with the experiments. This approach was then employed for proteins and peptides derived from an E. coli. cell lysate and digest. The results were compared to “traditional” proteomic approaches and increased coverage of trace proteins was obtained with the displacement approach. Finally, a salt-free ion exchange process was developed to permit direct coupling of displacement separations with on-line MS/MS analysis for peptide sequence determination.
 

Analytical Approaches: Bioanalytical Chemistry
1:00 PM-5:20 PM, Sunday, August 17, 2008 Courtyard by Marriott Downtown -- Logan, Oral

Division of Analytical Chemistry

The 236th ACS National Meeting, Philadelphia, PA, August 17-21, 2008