低场核磁共振分析蛋白质层析传质过程

Phase transfer was involved in every chromatographic step for the components in separation process. In this project we propose a new method using low-field NMR to analysis of the mass transfer in protein chromatography process. As low-field NMR can effectively measure the relaxation time of the hydrogen in solvents (small molecules), but is unable to detect that of the solutes (macromolecules), it can be used to detect the diffusion and mass transfer of solvents, and then analyze the mass transfer of protein in chromatographic medium. Firstly we can accurate determine the pore size and its distribution and the effects of cross-linking and ligand binding in different types of protein chromatographic medium, based on the preliminary research results of NMR relaxation time characterizations of water in the pores of the medium. Then deuterated water witch has no NMR signal can be used as a probe, to study the diffusion process of water and buffer solution in the pores of chromatographic medium. Finally, as the object of study, to in situ characterize chromatographic separation process, protein solutions containing or not heavy water will be added to the medium and tested by NMR, and then can be correlated to the medium properties. The research results of this project is expected to not only provide a better insight into mass transfer in protein chromatography, but also an important analysis method for designation and preparation of chromatographic media for protein separation.

层析过程的每一步骤中都涉及待分离组分在固液两相间的传递。本项目提出了一种用低场核磁共振分析蛋白质层析过程传质过程的的新方法。利用低场核磁的特性，即可以有效测定溶剂（小分子液体）氢的弛豫特征，但很难检测到溶质（大分子共价键合）的氢，从而可以在蛋白质层析中，通过溶剂的扩散传质过程进行蛋白质传质过程的分析。本项目首先在前期利用低场核磁共振对常规蛋白质层析介质孔内水弛豫特征研究结果的基础上，准确测定各种不同类型的层析介质孔径及其分布，分析各种交联和配基键合对层析介质结构特性的影响；然后以无核磁信号的重水作为探针，研究水及缓冲液在层析介质孔内的扩散过程；最后以蛋白质的水或重水溶液为对象检测蛋白质层析传质过程，并将其与层析介质特性相关联，用核磁共振进行层析分离的原位表征。本项目预期的研究成果不仅可以为蛋白质层析传质提供机理指导，并且可以为层析介质的设计和制备提供一个重要的分析手段。

层析过程的每一步骤中都涉及待分离组分在固液两相间的传递。本项目提出了一种用低场核磁共振分析蛋白质层析过程传质过程的的新方法。利用低场核磁的特性，即可以有效测定溶剂（小分子液体）氢的弛豫特征，但无法检测到溶质（大分子共价键合）的氢，从而可以在蛋白质层析中，通过溶剂的扩散传质过程推导蛋白质传质过程的分析。本项目首先在前期利用低场核磁共振对常规蛋白质层析介质孔内水弛豫特征研究结果的基础上，准确测定各种不同类型的层析介质孔径及其分布，分析了各种交联和配基键合对层析介质结构特性的影响。研究了具有表面延伸剂的层析介质的孔结构，建立了传质过程与层析介质特性的关系。研究并建立了介质原料水凝胶的胶凝点和溶胀行为测定的新方法。并研发了小型介质合成考察系统，准确控制了蛋白层析介质的孔结构，并与商品介质的层析传质特性进行了对比。本项目预期的研究成果不仅可以为蛋白质层析传质提供机理指导，并且可以为层析介质的设计和制备提供一个重要的分析手段。