The long term goal of this research is to develop, through advances in liquid chromatography, better tools for the fractionation and analysis of biological macromolecules. Five general areas will be examined; column technology, structure-retention relationships, protein structure analysis, high throughput preparative systems, and biospecific detectors. Research on column technology will be directed toward the development of stationary phase bonding chemistry for the preparation of chromatographic media on both non- porous and porous polystyrene supports that will be stable from pH 1 to 14 and be useful in both high speed analytical and preparative separations. Through the use of genetically created protein variants a further aim is to determine the relationship between chromatographic behavior and structural variables such as i) amino acid position on the protein surface, ii) three-dimensional structure of polypeptides under five kilodaltons, and iii) conformational changes.
The specific aim of the research on protein structural analysis is to use column switching techniques in i) chromatographic- immunological assays, ii) epitope mapping, and iii) the construction of psuedo-epitopes.
The final aims of the research are to develop preparative systems with a protein processing capability of 100 g/hr/L of column volume and biospecific detectors based on ligand displacement for monitoring the elution of specific proteins in analytical and preparative chromatography.
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