Novel Chromatofocusing Methods with Applications in Proteomics

University Of Maryland Baltimore County, Baltimore MD

Investigators

Abstract

Abstract Proposal Title: Novel chromatofocusing Methods with Applications in Proteomics Proposal Number: CTS-0442072 Principal Investigator: Douglas Frey Institution: University of Maryland College Park A new chromatographic technique, serial displacement chromatofocusing (SDC), will be developed and evaluated with regard to its usefulness as a replacement for preparative isoelectric focusing in the field of proteomics. To further enhance the usefulness of the method, theoretical tools involving computer-aided process design, the thermodynamics of protein adsorption, and bioinformatics, will be developed and employed. The proposed SDC method involves the formation of a self-generated, retained pH gradient consisting of a series of self-sharpening pH fronts, which propagate inside an ion-exchange column, which will use nanometer-sized particles as a packing material. This project will address three prototypical uses in the field of proteomics: (1) its use as an efficient prefractionation method prior in 2D gel electrophoresis, (2) its use as a rapid separation method for use in high-throughput proteomics, and (3) its use as a non-denaturing first separation dimension in a 2D zymogram method for detecting enzymatic activity. In terms of the broader impacts, the work has applications in several areas of critical national importance, such as in the search for new diagnostic markers for diseases and in the rapid identification of microorganisms for food safety and counter-bioterrorism. In addition, the proposed work is an interdisciplinary study that involves aspects of chromatographic process engineering, bioanalytical chemistry, and applied biology, so that students working on the project will be well prepared to join the U.S. workforce in the biotechnology area. Through the Meyerhoff scholarship program, students at both the graduate and undergraduate levels will be recruited to work on this project, which will help to increase the number minorities working in the U.S. biotechnology industry.