US-West Germany Cooperative Research in Kinetic Effects in Protein Chromatography

该奖项支持特拉华大学的Abraham M.Lenhoff教授与德意志联邦共和国美因茨的Johannes Gutenberg大学无机和分析化学研究所的Klaus K.Unger教授进行合作研究。他们研究的目的是为了更好地了解吸附和解吸动力学在蛋白质疏水相互作用层析(HIC)中的作用。所寻求的信息对分析和制备系统都具有重要意义。它为微量分析分离的速度设定了一个基本限制，并提供了速率信息，这些信息对于优化这种形式的色谱柱的制备柱的设计和操作可能是重要的。将使用两种互补的分析方法，两种方法都涉及相同的蛋白质、表面特征和溶剂组成。第一个涉及使用全内反射荧光(TIRF)光谱来直接、实时地测量平板载玻片上的吸附和解吸动力学和平衡。第二种方法使用带有1.5微米非多孔固定相的HIC来检测实际柱层析中的行为，这些条件下的动力学可能是限速的。将通过对各自系统的传输和动力学建模来比较两组实验的结果。计划中的合作将特拉华集团的TIRF能力和建模优势与美因茨集团的表面合成专业知识结合在一起。

This award supports Professor Abraham M. Lenhoff of the University of Delaware for collaborative research with Professor Klaus K. Unger of the Institute for Inorganic and Analytical Chemistry of Johannes Gutenberg University, Mainz, Federal Republic of Germany. The objective of their research is to gain a better understanding of the role of adsorption and desorption kinetics in hydrophobic interaction chroma- tography (HIC) of proteins. The information sought is significant for both analytical and preparative systems. It sets a fundamental limit for the speed of microanalytical separations, and it provides rate information that can be important in optimizing the design and operation of preparative columns for this form of chromatography. Two complementary analytic approaches will be used, both involving the same proteins, surface characteristics and solvent compositions. The first involves the use of Total Internal Reflectance Fluorescence (TIRF) spectroscopy for the direct, real-time measurement of adsorption and desorption kinetics and equilibria on a flat slide. The second uses HIC with a 1.5 micron non-porous stationary phase to examine behavior in actual column chromatography under conditions where kinetics are likely to be rate-limiting. Comparison of the results of the two sets of experiments will be made possible by the transport and kinetic modelling of the respective systems. The planned cooperation combines the TIRF capability and the modelling strengths of the Delaware group with the surface synthesis expertise of the Mainz group.