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约翰内斯古腾堡大学无机和分析化学研究所的Klaus K. Unger教授的合作研究。他们的研究目的是为了更好地理解蛋白质的疏水相互作用色谱（HIC）中吸附和解吸动力学的作用。所寻求的资料对分析系统和准备系统都很重要。它为微分析分离的速度设定了一个基本限制，并且它提供的速率信息对于优化这种形式的色谱制备柱的设计和操作是重要的。将使用两种互补的分析方法，两者都涉及相同的蛋白质，表面特性和溶剂组成。第一个涉及使用全内反射荧光（TIRF）光谱直接，实时测量吸附和解吸动力学和平衡在一个平坦的幻灯片。第二种方法使用HIC与1.5微米的无孔固定相，在动力学可能限速的条件下检查实际柱色谱中的行为。通过对各自系统的输运和动力学建模，可以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相结合。

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.