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 FERNANDEZTHETHIS NSF颁发的化学和生物学分离计划支持Erik Fernandez教授，Michael Shirts和John O'Connell教授的工作，以分析和预测色谱纯化期间的蛋白质结构变化。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。主要焦点是疏水相互作用（HIC），尤其是离子交换色谱法（IEC），即将所需蛋白质与化学改性，高分子量和构象变体分开的重要商业方法。研究的目的是1）在蛋白质吸附到HIC和IEC媒体期间的全球和局部不稳定性的分子细节，2）开发一种方法，以预测操作条件对结构和物种分布的影响，以及3）通过我们的测试预测单一和多域蛋白质中构象变化的结构和色谱后果的工具。 该项目将添加分子建模工具，以实现对更复杂，更大（？多域？）蛋白质的分析。结果应使表面和蛋白质的先验设计能够增强HIC和IEC的选择性和产量，以促进药物相关蛋白（例如抗体）。 将创建有关生物分子系统动力学和平衡的教育模块，以用于本科生物化学工程和热力学课程，包括各种计算练习和可视化。化学工程师与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的技术。 与两家生物技术公司的合作将使学生面临蛋白质稳定性和加工的复杂，与商业上相关的问题，并提供与药物相关的多域蛋白质。开发的仿真工具将是开源的，并在线发布以供其他研究人员访问。