Separation of Macromolecules Based on Conformation Pathways

这项由美国国家科学基金会化学和生物分离项目颁发的奖项支持了布朗大学Anubhav Tripathi教授探索分离和/或定量这种“相似”物质的新方法的工作。在完全折叠或未折叠状态下具有相似迁移率的两个同工异构体之间产生构象差异的条件下进行分离的大分子。研究工作利用微流控筛选平台确定了大分子构象变化对溶剂质量的依赖关系，以确定最佳缓冲液。该研究项目还开发了一种新的微流控系统，用于在高盐的最佳缓冲液中分离异构体。最后，研究工作开发了数值和分析工具来分析具有改变构象的紧密迁移性大分子的分离。这项工作的结果可能会导致分离和纯化科学的一个新的研究方向，能够通过利用微量样品分离和/或定量同种异构体，并可能导致对所提出的方法分离的机制理解。鉴于具有类似流动性的大分子在突变、不适当表达或功能（例如朊病毒和癌症疾病）的病理状态中的核心作用，它们的量化变得非常重要。除了临床应用外，这项研究工作还可以帮助建立一个“图书馆”。各种蛋白质的构象特征，这可以帮助研究人员探索蛋白质折叠的复杂问题。最后，将热力学、动力学和输运现象的核心技能与分离和净化科学相结合。该计划支持开发运输过程中的新课程。该计划为探索微流体的新本科实验室提供了极好的机会。生物技术和纳米技术行业的基础工程研究和分析开发之间的紧密联系得到了促进。

CBET-0756588TripathiThis NSF award by the Chemical and Biological Separations program supports work by Professor Anubhav Tripathi at Brown University to explore a novel approach for separation and/or quantification of such ?similar? macromolecules by performing separation under conditions in which conformations differences arise between two isoforms that have similar mobilities in completely folded or unfolded states. The research work determines the dependence of conformation changes of macromolecules on solvent quality to identify the optimal buffer using a microfluidic screening platform. The research program also develops a new microfluidic system for separation of the isoforms in the optimal buffers with high salts. Lastly, the research work develops numerical and analytical tools to analyze separation of close mobility macromolecules with altered conformations. The results of this work is likely to result in a new research direction in separation and purification science, capable of separating and/or quantifying isoforms by utilizing minute amounts of sample, and can lead to a mechanistic understanding of the proposed approach separation. Quantification of macromolecules with similar mobilities has become important in light of their central role in pathological states of mutations, improper expression or function (e.g. prion and cancer diseases). In addition to its clinical utility, this research work can assist in development of a ?library? of conformational signatures of various proteins, which could help researchers that are exploring the complex problem of protein folding. Finally, the educational program couples core skills of thermodynamics, kinetics and transport phenomenon to separation and purification science. The program supports development of new courses in transport processes. The program offers excellent opportunities to new undergraduate laboratories exploring microfluidics. Strong ties are promoted between the fundamental engineering research and assay development in biotechnology and nanotechnology industries.