Novel, High Performance Detection for Bioseparations

Koropchak, John

Abstract

The overall objective of the proposed research is to develop simple, practical methods of analysis which can be applied to biochemical problems to provide faster, more comprehensive, and sensitive detection. Separation methods including HPLC and CE will be coupled to a new detection system (condensation nucleation light scattering ) in order to provide a simple, practical method of analysis applicable to biochemical problems. The new detection system is nearly universal (i.e., only requiring low volatility for detection) , and theoretically it is capable of detection even to the single molecule level. The project focuses on the development of methodology with applications in biochemistry, biotechnology and biomedicine. In particular, separations and detection of underivatized amino acids, peptides and proteins will be developed and characterized with both HPLC and CE, including application to peptide mapping of native authentic and recombinant proteins. It is anticipated that these new methods will provide more accurate and comprehensive information, which will minimize the health risks that may result from the utilization of these materials. Other efforts will be devoted to the development of methods for the analysis of mycotoxins called fumonisins, which are produced by the most common fungi in corn. Current methods are not amenable to widespread utilization and identification of these toxins; it is anticipated that the methods developed by this work will alleviate that difficulty. Other applications include the measurement of phospholipids; however, utilization of the methodology developed in many other areas can be envisioned.