Integrated Program for Conformational Effects in Protein Chromatography

蛋白质色谱构象效应综合程序

• 批准号: 1134256
• 负责人: Giorgio Carta
• 金额: $ 30万
• 依托单位: University of Virginia Main Campus
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2016-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1134256&HistoricalAwards=false
• 关键词: Integrated; Program; Conformational; Effects; Protein

1134256FernandezThis NSF award by the Chemical and Biological Separations program supports work by Professors Erik Fernandez, Michael Shirts, and John O'Connell to analyze and predict protein structural changes during purification by chromatography. This will be accomplished by combining extensive experimental measurements, detailed molecular simulations, and thermodynamic modeling. The principal focus is hydrophobic interaction (HIC) and especially ion-exchange chromatography (IEC), vital commercial methods to separate desired proteins from chemically modified, high molecular weight, and conformational variants. The aims of the research are to 1) establish molecular details of global and local instabilities during protein adsorption to HIC and IEC media, 2) develop a methodology for predicting effects of operating conditions on structural and species distributions, and 3) test predictions with our tools for structural and chromatographic consequences of conformation changes in single- and multidomain proteins. This project will add molecular modeling tools to enable analysis of more complex, larger (?multidomain?) proteins. The outcomes should enable a priori design of surfaces and proteins to enhance selectivity and yield in both HIC and IEC for pharmaceutically relevant proteins such as antibodies. An educational module on dynamics and equilibria of biomolecular systems will be created for use in undergraduate biochemical engineering and thermodynamics courses, including a variety of computational exercises and visualizations. Collaboration between the chemical engineers and a prominent protein-folding researcher, will enable students to work with advanced techniques on both fundamental and practical problems. Collaboration with two biotechnology companies will expose students to complex, commercially-relevant issues of protein stability and processing and provide with pharmaceutically relevant multidomain proteins. Simulation tools developed will be open-source and posted on-line for access by other researchers.

1134256费尔南德斯这项由化学和生物分离计划颁发的NSF奖支持埃里克·费尔南德斯、迈克尔·衬衫和约翰·奥康奈尔教授在通过层析纯化过程中分析和预测蛋白质结构变化的工作。这将通过结合广泛的实验测量、详细的分子模拟和热力学建模来实现。主要的焦点是疏水相互作用(HIC)，特别是离子交换层析(IEC)，这是从化学修饰、高分子量和构象变体中分离所需蛋白质的重要商业方法。这项研究的目的是1)建立蛋白质在HIC和IEC介质上吸附过程中全局和局部不稳定性的分子细节，2)开发一种预测操作条件对结构和物种分布的影响的方法，以及3)使用我们的工具测试单域和多结构域蛋白质构象变化的结构和层析结果。该项目将增加分子建模工具，以实现对更复杂、更大(多域)的分析。蛋白质。这些结果应该能够对表面和蛋白质进行先验设计，以提高抗体等药物相关蛋白质在HIC和IEC中的选择性和产率。生物分子系统的动力学和平衡的教育模块将被创建用于本科生的生物化学工程和热力学课程，包括各种计算练习和可视化。化学工程师和一位著名的蛋白质折叠研究人员之间的合作，将使学生能够在基本和实际问题上使用先进技术。与两家生物技术公司的合作将使学生接触到复杂的、与商业相关的蛋白质稳定性和加工问题，并提供与药物相关的多域蛋白质。开发的模拟工具将是开源的，并发布在网上，供其他研究人员访问。