Capillary-Channeled Polymer Fibers: Chemistries and Structures for Protein Separations and Analytics

在化学测量和成像项目（化学部）的支持下，R. Kenneth Marcus教授、他的合作者和克莱姆森大学的学生正在开发和表征毛细管通道聚合物（C-CP）纤维作为快速处理生物大分子的固定相。C-CP材料适用于广泛的柱格式（微孔到制备），并且相对于传统的多孔球，膜和整体固定相提供了大分子分离的优势，包括低流动阻力，最小化尺寸排斥效应，更高的传质速率，高流动相速度（不一定是体积流速）和梯度速率，而不牺牲色谱性能。拟议的研究是针对开发C-CP纤维柱的基本特征，以影响蛋白质分析领域的高产和选择性分离。这项工作必然是跨学科的，涉及分析和生物化学、化学工程和纺织科学研究人员之间的合作。将采取双管齐下的办法。第一个研究领域将集中在修饰表面化学物质以影响高水平的选择性。这些修饰不仅要评估它们的化学选择性，还要评估配体的利用效率、对非特异性结合的敏感性，以及在过程循环方面的稳健性（包括苛性清洁程序）。在所有情况下，性能指标将直接与商业上可用的固定相填充的列进行比较。第二个研究领域超越了色谱柱结构，涉及损耗/捕获柱和微移管尖端结构的流体动力学方面的表征和优化，以及它们分别如何影响在线和离散样品蛋白质处理的性能。正如科学家为研究生物的生物学功能而编目生物体DNA中的所有基因一样，他们也在发展蛋白质分析，最终能够编目生物体用于实现其功能的不同蛋白质集合。完整、准确的蛋白质目录对于开发更有效的药物、诊断工具和分子医学的进步至关重要。他们严重依赖于高效的分离技术，因为生物样本非常复杂。Marcus教授的这项工作将通过开发分离技术来推进蛋白质分析，该技术可以在更短的时间内更准确地分析更复杂的样品。这些研究涉及化学系和材料科学与工程学院的研究小组之间的合作努力，这两个研究小组都隶属于克莱姆森大学纤维和薄膜高级工程中心（CAEFF）。与联邦实验室的合作也在计划之中。这项工作的成功完成可能会导致南卡罗来纳州和其他地方的纺织和制药行业出现新的商业风险。CAEFF项目让研究生接触科学和研究哲学，跨越科学/工程界面。构建C-CP纤维柱的低成本和简单性将允许它们在新设计的本科生和研究生实验室实验中使用，说明蛋白质色谱的基本原理。

With support from the Chemical Measurement and Imaging Program (Division of Chemistry), Professor R. Kenneth Marcus, his collaborators and their students at Clemson University are developing and characterizing capillary-channeled polymer (C-CP) fibers as stationary phases for rapid processing of biomacromolecules. C-CP materials are amenable to a wide range of column formats (microbore to preparative) and offer advantages for macromolecular separations relative to traditional porous-bead, membrane, and monolithic stationary phases, including low flow resistance, minimization of size-exclusion effects, much higher mass transfer rates, and high mobile phase velocities (not necessarily volume flow rates) and gradient rates without sacrificing chromatographic performance. Proposed studies are directed at exploiting the basic characteristics of C-CP fiber columns to affect highly productive and selective separations in the realm of protein analytics. This work is necessarily interdisciplinary, involving cooperation between analytical and biochemistries, chemical engineering, and textile science researchers. A two-pronged approach will be taken. The first research area will focus on modification of surface chemistries to affect high levels of selectivity. The modifications will be evaluated not only for their chemical selectivity, but also the ligand utilization efficiency, susceptibility to non-specific binding, and robustness in terms of process cycling (including caustic clean in place procedures). In all cases, performance metrics will be directly compared to columns packed with commercially available stationary phases. The second research area looks beyond chromatographic column structures, involving the characterization and optimization of the hydrodynamic aspects of depletion/capture columns and micropipette tip constructs and how they affect performance of in-line and discrete sample protein processing, respectively.Just as scientists catalog all the genes in an organism's DNA to study its biological function, they are also developing protein analytics to eventually be able to catalog the varying collections of proteins an organism uses to carry out its functions. Complete, accurate protein catalogs are critical to the development of more effective pharmaceuticals, diagnostic tools and the advance of molecular medicine. They rely heavily on efficient separations technology because biological samples are so complex. This work conducted by Prof. Marcus will advance protein analytics by developing separations technology that can analyze more complex samples more accurately in less time. The studies involve a collaborative effort between research groups from the Department of Chemistry and the School of Materials Science and Engineering, both of which are affiliated with the Clemson University Center for Advanced Engineering of Fibers and Films (CAEFF). Collaborations with federal laboratories are also planned. Successful completion of the work may lead to new commercial ventures in the textile and pharmaceutical industries in South Carolina and elsewhere. CAEFF programs expose graduate students to science and research philosophies across the science/engineering interface. The low cost and simplicity of constructing C-CP fiber columns will allow their use in newly designed undergraduate and graduate laboratory experiments illustrating the basic principles of protein chromatography.