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

提案编号：0730830主要分析员：Steven Cramer机构：Rensselaer Polytechnic InstituteProPOSAL Title：蛋白质纯化的化学选择性置换剂蛋白质、置换剂结构、移动的相特性和置换剂浓度对置换色谱系统的有效性和选择性的影响将使用自动液体处理系统进行检查，以实现多维筛选。将合成从高通量筛选（HTS）研究中选择的高亲和力和选择性置换剂先导化合物。在整个合成工作中，将研究将发色团和/或荧光团引入这些置换剂的各种方法，以便于在线检测。这项工作将检查置换剂，有能力结合到特定类别的蛋白质。 除了从HTS研究中确定的选择性置换剂之外，这项工作还将研究使用已知的结合配体拴系到适当带电部分的类选择性置换剂的先验设计。在该项目的这一阶段开发的置换剂将被评估其在批处理和柱实验下选择性置换所需蛋白质的能力。选择性置换剂和蛋白质之间的相互作用模式也将使用多种技术进行研究。 基于空间质量作用等温线的四态模拟模型将用于预测这些选择性置换剂的柱性能。拟议的研究将对大规模生物加工和分析生物技术产生影响。开发与特定蛋白质或蛋白质类别结合同时也充当有效置换剂的选择性置换剂将显著增加离子交换系统的选择性。拟议的工作可能对参与细胞调控的关键蛋白质的纯化产生重大影响。 建议的研究也将有一个重要的影响，研究生和本科化学工程专业的学生的教育。参与这个多学科项目的学生将获得色谱生物处理，建模，机器人和各种分析技术，如毛细管区带电泳，表面等离子体共振和NMR的重要培训。本计画之研究成果将纳入层析分离制程之课程中。 最后，在PI实验室实习的高中生也将获得重要的实验室经验。 如果成功，该提案将对生物加工和许多与健康相关的领域产生重大而广泛的影响。

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.