Use of displacement chromatography for proteomic applications


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
Proteomics provides one of the most important bioanalytical challenges of our time. Displacement chromatography has significant potential for the simultaneous concentration and purification of complex biological mixtures. Model studies were carried out using high affinity, low molecular mass displacers for the ion exchange displacement chromatography of proteins and peptides to determine the limits of this approach. Parametric studies with various displacers, displacer concentrations, stationary phases, and mobile phase conditions were performed to optimize these difficult separations. Results indicate high affinity displacers employed at relatively low concentrations can effect high resolution separations amenable to the identification of low abundance solutes. Displacement ion exchange systems were then used in concert with nano-flow reversed phased liquid chromatography and mass spectrometry to evaluate the utility of this combined approach for the analysis of complex biological mixtures with a wide dynamic range of feed concentrations.