Fundamentals of Protein Transport in Charged Gels for Chromatography and Membrane Applications

CTS-0079334 Giorgio CartaUniversity of Virginia色谱和膜应用中带电凝胶中蛋白质转运的基础摘要本项目研究了为色谱和膜应用制备的带电水凝胶中的蛋白质转运。重点放在支持凝胶通过在多孔基质和其他刚性结构中原位合成它们。目前正在研究以下性质：（一）支持的离子凝胶的尺寸排阻、流体动力学渗透性和电性质;（二）通过同位素交换和放射自显影方法测量离子凝胶中蛋白质和电解质的分配和迁移率;和（iii）单-蛋白质吸附和多组分系统在离子凝胶稳定在毛细管和多孔颗粒使用两种直接显微镜可视化方法和宏观测量。 实验数据被用来开发速率模型来描述蛋白质在离子凝胶中的传输，速率理论被纳入柱色谱模型中，以预测和优化实际介质的分离性能。另一个目标是开发新的膜的基础上支持离子凝胶，以实现蛋白质separation.The总体目标，这项研究是一个基本的了解蛋白质运输带相反电荷的离子聚合物水凝胶作为基础，合理开发有效的媒体生物技术的应用。聚合物水凝胶在该领域具有许多当前和潜在的用途。虽然中性水凝胶用作尺寸排阻介质，但结合到聚合物主链的固定电荷的结合引入了从稀溶液可逆地积累大分子的能力。因此，离子水凝胶可用于蛋白质分离的色谱和膜过程，用于药物的透皮递送，作为治疗剂长期递送的可植入载体，以及作为诊断仪器中的元件。 了解带电大分子的输运性质是优化设计这些材料的关键。这项研究也将通过学生的参与对教育和人力资源开发产生重大影响，并将促进生物技术的发展。分离和纯化通常被认为是工业生物加工中最昂贵的步骤。因此，开发改进的色谱固定相和分离膜以及对该领域的科学家和工程师进行教育至关重要。

CTS-0079334Giorgio CartaUniversity of Virginia Fundamentals of Protein Transport in Charged Gels for Chromatography and Membrane ApplicationsAbstractThis project examines protein transport in charged hydrogels prepared for chromatography and membrane applications. The focus is placed on supported gels made by synthesizing them in situ within porous matrices and other rigid structures. The following properties are being investigated: (i) Size exclusion, hydrodynamic permeability, and electrical properties of supported ionic gels; (ii) Protein and electrolyte partitioning and mobility in ionic gels measured through isotope exchange and autoradiography methods; and (iii) Interdiffusion rates for single-protein adsorption and for multicomponent systems in ionic gels stabilized in capillaries and in porous particles using both direct microscopic visualization methods and macroscopic measurements. The experimental data are being used to develop rate models to describe transport of proteins in ionic gels, and the rate theories are incorporated in models for column chromatography to predict and optimize separation performance with practical media. An additional goal is to develop novel membranes based on supported ionic gels to effect protein separations.The overall goal of this research is a fundamental understanding of protein transport in oppositely charged ionic polymeric hydrogels to serve as a basis for the rational development of effective media for biotechnological applications. Polymeric hydrogels have many current and potential uses in this field. While neutral hydrogels serve as size-exclusion media, incorporation of a fixed charge, bound to the polymer backbone introduces the ability to accumulate macromolecules reversibly from dilute solutions. Ionic hydrogels can thus be used in chromatographic and membrane processes for protein separations, for the transdermal delivery of drugs, as implantable vehicles for the long-term delivery of therapeutic agents, and as elements in diagnostic instruments. Understanding the transport properties of charged macromolecules is a key to the optimum design of these materials. The research will also have significant impact on education and development of human resources through the involvement of students and will enhance the development of biotechnology. Separation and purification are generally thought to be the costliest steps in industrial bioprocessing. Hence, the development of improved chromatographic stationary phases and membranes for separations and the education of scientists and engineers in this field are crucial.