Novel Adsorbents and Transport Mechanisms for Enhanced Protein Mass Transfer in Ion Exchange Chromatography

用于增强离子交换色谱中蛋白质传质的新型吸附剂和传输机制

• 批准号: 0729857
• 负责人: Giorgio Carta
• 金额: $ 25万
• 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: 2007
• 资助国家: 美国
• 起止时间: 2007-11-01 至 2010-10-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0729857&HistoricalAwards=false
• 关键词: Novel; Adsorbents; Transport; Mechanisms; Enhanced

PROPOSAL NUMBER: 0730830PRINCIPAL INVESTIGATOR: Steven CramerINSTITUTION: Rensselaer Polytechnic InstitutePROPOSAL TITLE: Chemically Selective Displacers for Protein PurificationThe effects of protein, displacer structure, mobile phase characteristics and displacer concentration on the efficacy and selectivity of displacement chromatography systems will be examined using an automated liquid handling system to enable multi-dimensional screens. High affinity and selective displacer lead compounds selected from the high throughput screening (HTS) studies will be synthesized. Throughout this synthesis work various approaches to introduce chromophores and/or fluorophores into these displacers will be examined in order to facilitate on-line detection. This work will examine displacers that have the ability to bind to specific classes of proteins. In addition to selective displacers identified from the HTS studies, this work will investigate the a priori design of class selective displacers using known binding ligands tethered to appropriately charged moieties. Displacers developed in this phase of the project will be evaluated for their ability to selectively displace the desired proteins under batch and column experiments. The mode of interaction between selective displacers and proteins will also be investigated using multiple techniques. A four state simulation model based on the steric mass action isotherm will be used to predict column performance of these selective displacers. The proposed research will have an impact on both large scale bioprocessing as well as analytical biotechnology. The development of selective displacers that bind to a specific protein or protein class while also acting as efficient displacers will dramatically increase the selectivity of ion exchange systems. The proposed work could have a major impact on the purification of key proteins involved in cellular regulation. The proposed research will also have an important impact on the education of both graduate and undergraduate chemical engineering students. The students involved in this multidisciplinary project will obtain significant training in chromatographic bioprocessing, modeling, robotics, and a variety of analytical techiques such as capillary zone electrophoresis, surface plasmon resonance and NMR. The research from this project will be incorporated into a course on Chromatographic Separation Processes. Finally, high school students who are carrying out internships in the PI's laboratory will also obtain important laboratory experience. If successful the proposal will have a significant and broad impact on bioprocessing and a number of health related fields.

建议编号：0730830原理研究员：史蒂文cramerinstitution：Rensselaer Polytechnic Institutpropropropropropropropropropolopolopolososal标题：用于蛋白质纯化的化学选择性位移蛋白纯化蛋白质，位移结构，流动相位特征和置换液的液体功能的蛋白质的影响，临床相结构和位移层的液体系统会被检查系统以实现多维屏幕。将合成从高吞吐量筛选（HTS）研究中选择的高亲和力和选择性置换铅化合物。在整个综合工作中，将检查各种将发色团和/或荧光团引入这些位移器的方法，以促进在线检测。这项工作将检查具有结合特定类别蛋白质的能力的位移。 除了从HTS研究中鉴定出的选择性位移外，这项工作还将使用已知的结合配体束缚以适当充电的部分研究类选择性位移的先验设计。将在项目的这一阶段开发的位移器，以评估其在批处理和柱实验下选择性位移所需蛋白质的能力。选择性位移和蛋白质之间的相互作用方式也将使用多种技术研究。 基于空间质量作用等温线的四个状态仿真模型将用于预测这些选择性位移的列性能。拟议的研究将对大规模生物处理和分析生物技术产生影响。与特定蛋白质或蛋白质类结合的选择性位移剂的发展，同时充当有效的位移剂也将显着提高离子交换系统的选择性。提出的工作可能会对参与细胞调节的关键蛋白的纯化产生重大影响。 拟议的研究还将对研究生和本科化学工程专业学生的教育产生重要影响。参与这个多学科项目的学生将获得色谱生物处理，建模，机器人技术以及各种分析技术（例如毛细管电泳，表面等离子体等离子体共振和NMR）的重大培训。该项目的研究将纳入色谱分离过程的课程中。 最后，在PI实验室进行实习的高中生也将获得重要的实验室经验。 如果成功，该提案将对生物加入和许多与健康相关的领域产生重大且广泛的影响。