Solute Transport and Binding Kinetics in Affinity Chromatography

一种新型光纤探针将用于研究：(1)溶质通过载体基质的运动；(2)固定化配体结合溶质的内在动力学。探针为光纤，利用倏逝波在光纤表面激发分子；作为波的电磁场仅能穿透2000埃进入周围介质，可以用来研究光纤表面的分子。在该项目的第二部分，波导将被覆盖一层非常薄的支撑矩阵薄膜，并确定感兴趣的溶质通过亲和柱的运动速率。这将允许通过直接测量分离传质和平衡效应。因此，支持矩阵的性质和配体连接的方法的影响可以明确地确定。这些知识对于亲和、色谱、生物技术中应用的分离技术将是相当重要的。亲和分离是生物制品回收的一项重要技术。虽然研究广泛，但大多数研究都停留在宏观层面；例如，一些研究使用了色谱理论中的正面分析或脉冲技术。本项目将尝试从微观角度研究影响结合常数和传质速率的因素。

A novel fiber optic probe will be developed and then used to investigate: (1) the movement of solutes through a support matrix and (2) the intrinsic kinetics of solute binding by immobilized ligands. The probe is an optical fiber employing an evanescent wave to excite molecules at the fiber surface; the electromagnetic field which is the wave penetrates only 2000 angstroms into the surrounding medium, and can be used to study the molecules at the surface of the optical fiber. In the second part of the project, the waveguide will be coated with a very thin film of a support matrix and the rate of movement of solutes of interest through an affinity column will be determined. This will allow separation of mass transfer and equilibrium effects by direct measurement. Thus, the influence of the nature of the support matrix and the method of ligand attachment can be unambiguously determined. This knowledge will be of considerable importance in affinity, chromatography, a separation technique with applications in biotechnology. Affinity-based separations are an important technique for the recovery of biological products. Though extensively studied, most investigations have been at the macroscopic level; for example, some studies have used frontal analysis or pulse techniques from chromatography theory. This project will attempt a study of factors affecting binding constants and mass transfer rates from a microscopic perspective.