LIQUID CHROMATOGRAPHY OF BIOLOGICAL SUBSTANCES

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This proposal is aimed at expanding the scope of high performance liquid chromatography and its application to medicine and the life sciences along several congruos lines. The molecular interactions involved in the retention of biopolymers by stationary phases in surface mediated chromatographic processes will be investigated in detail. The results of this study on hydrophobic, electrostatic and biospecific interactions are expected to shed light on the relevant surface phenomena in biological systems and to provide a means for the characterization of various stationary phases used in practice. A major research effort will be undertaken to develop highly efficient chromatographic separation techniques for glycans and their conjugates in order to meet an acute need for suitable tools for analysis and purification of such substances, whose biological significance is being increasingly recognized. Concurrently, efficient methods will be developed for the isolation and purification of lectins and oligosaccharides to be used in conjunction with our research on chromatography of biological carbohydrates. Systematization of these procedures will permit the establishment of a set of unit operations for the isolation of complex molecules from biological matrices. Experience gained in the course of this project will be used in ad hoc collaborative research projects on separation problems of importance to the life sciences and medicine. Another part of the proposed research will focus on the rapid analysis of proteins and nucleic acid on columns packed with micropellicular sorbents and using appropriately modified instrumentation. This approach promises at least a ten-fold reduction in the time of analysis. Novel stationary phases based on macroporous siliceous and polymeric microspheres will be developed for the analysis of proteins, nucleic acids and biologically important sugars; the experience gained in the analytical chromatography of these substances will be used to prepare sorbents for their preparative separation. Studies on the theory and practice of non-linear elution chromatography are expected to provide a framework for the analysis and design of the preparative chromatographic process.