Displacer concentration effects in displacement chromatography: Proteomic applications

BIOT 299

Steven Taylor Evans, evanss3@rpi.edu1, Alexander S. Freed, freeda@rpi.edu1, Amy Manocchi1, and Steve M. Cramer, crames@rpi.edu2. (1) Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, Troy, NY 12180, (2) Chemical and Biological Engineering, Rensselaer Polytechnic Institute, 110 8th Street, Ricketts Bldg. 130, Troy, NY 12180
Displacement chromatography can simultaneously purify and concentrate solutes making it attractive for proteomic applications. Increased dynamic range requires low abundant protein detection in the presence of high abundance proteins having similar affinities for chromatographic resins. In order to examine this, a model study was carried out using high affinity, low molecular mass displacers to determine the limits of this approach for protein purification in ion exchange systems. Model simulations were also carried out using a Steric Mass Action based chromatographic model to study the correlation between relative feed concentrations, separation factors, displacer concentration, and salt concentration on the yield, purity, shock layer, and detection of solutes present in low concentrations. Results indicate high affinity displacers employed at relatively low concentrations can effect high resolution separations amenable to the identification of low abundance proteins. Implications for displacement chromatography as a complementary multidimensional separation tool and mass spectrometry interfacing will be discussed.


Poster Session
5:30 PM-7:30 PM, Wednesday, 13 September 2006 Moscone Center -- Hall D, Poster

8:00 PM-10:00 PM, Monday, 11 September 2006 Moscone Center -- Hall D, Sci-Mix

Division of Biochemical Technology

The 232nd ACS National Meeting, San Francisco, CA, September 10-14, 2006