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.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM025431-11
Application #
3272991
Study Section
Metallobiochemistry Study Section (BMT)
Project Start
1978-07-01
Project End
1993-06-30
Budget Start
1988-07-01
Budget End
1989-06-30
Support Year
11
Fiscal Year
1988
Total Cost
Indirect Cost
Name
Purdue University
Department
Type
Earth Sciences/Resources
DUNS #
072051394
City
West Lafayette
State
IN
Country
United States
Zip Code
47907
Xu, W; Regnier, F E (1999) Electrokinetically-driven cation-exchange chromatography of proteins and its comparison with pressure-driven high-performance liquid chromatography. J Chromatogr A 853:243-56
Paliwal, S K; de Frutos, M; Regnier, F E (1996) Rapid separations of proteins by liquid chromatography. Methods Enzymol 270:133-51
Ratnayake, C K; Regnier, F E (1996) Study of protein binding to a silica support with a polymeric cation-exchange coating. J Chromatogr A 743:15-23
Ratnayake, C K; Regnier, F E (1996) Lateral interaction between electrostatically adsorbed and covalently immobilized proteins on the surface of cation-exchange sorbents. J Chromatogr A 743:25-32
Patterson, D H; Harmon, B J; Regnier, F E (1994) Electrophoretically mediated microanalysis of calcium. J Chromatogr A 662:389-95
Regehr, M F; Paliwal, S K; Regnier, F E (1994) Ensemble averaging and digital filtering in chromatography and electrophoresis. J Chromatogr A 659:247-53
Miller, K J; Leesong, I; Bao, J et al. (1993) Electrophoretically mediated microanalysis of leucine aminopeptidase in complex matrices using time-resolved laser-induced fluorescence detection. Anal Chem 65:3267-70
Wu, D; Regnier, F E (1993) Native protein separations and enzyme microassays by capillary zone and gel electrophoresis. Anal Chem 65:2029-35
Harmon, B J; Patterson, D H; Regnier, F E (1993) Electrophoretically mediated microanalysis of ethanol. J Chromatogr A 657:429-34
de Frutos, M; Paliwal, S K; Regnier, F E (1993) Liquid chromatography based enzyme-amplified immunological assays in fused-silica capillaries at the zeptomole level. Anal Chem 65:2159-63

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