Displacement Chromatography of Proteins: Identification of Critical Displacer Properties and Development of Novel Nontoxic Displacers

该项目研究了驱替色谱和聚合物合成，以研究使用定义明确的聚合物驱替剂化合物的关键驱替剂性能的影响。多种新型聚合物取代剂将被合成用于离子交换、疏水相互作用和金属亲和色谱系统。所开发的置换剂将进行纯化，并采用适当的分析化学技术来确定其结构和纯度。这些化合物的色谱行为将用洗脱色谱法进行评价，吸附等温线将用正面色谱法确定。将为这些吸附剂系统建立具有一系列特性的蛋白质饲料混合物模型。我们将研究不同分子量、官能团密度和构象的聚合物取代剂在这些体系中作为蛋白质取代剂的能力，并确定这些性质的相对重要性。有关驱替剂性能的所得数据将用于确定驱替剂物理化学性能的组合，这是生物聚合物的有效驱替和固定相材料再生所需的驱替剂的快速去除。这项基础研究将为分子水平上的位移现象提供基本的见解，并将为未来生物分离问题的设计和合成适当的位移剂建立指导方针。

The project studies displacement chromatography and polymer synthesis to investigate the effect of critical displacer properties using well defined polymeric displacer compounds. A variety of novel polymeric displacers will be synthesized for ion exchange, hydrophobic interaction, and metal affinity chromatographic systems. The developed displacers will be purified and subjected to appropriate analytical chemical techniques to establish their structure and purity. Chromatographic behavior of these compounds will be evaluated using elution chromatography and adsorption isotherms will be determined by frontal chromatography. Model protein feed mixtures exhibiting a spectrum of properties will be established for these adsorbent systems. The ability of polymeric displacers of various molecular weights, functional group densities, and conformations to act as protein displacers in these systems will be examined, and the relative importance of these properties will be established. The resulting data on displacer properties will be used to identify combinations of displacer physicochemical properties which are required for efficient displacement of biopolymers and rapid removal of the displacers for regeneration of the stationary phase materials. This fundamental research will provide basic insight into the displacement phenomenon at the molecular level and will establish guidelines for the design and synthesis of appropriate displacers for future bioseparation problems.