Protein Purification Using Novel Methods for Buffer Focusing Chromatography

摘要本基金由化学与传输系统部界面、传输与分离项目的分离与净化项目子项目授予。首席研究员是马里兰大学巴尔的摩分校的道格拉斯·弗雷博士。拟议的研究涉及研究技术，以有效地纯化蛋白质，而不将它们暴露在可能变性或沉淀的pH范围内。这些方法通常称为缓冲聚焦色谱法或聚焦色谱法。它们目前没有大规模使用，因为从业者对如何使用低成本的缓冲剂在低离子强度下形成具有高缓冲能力的保留pH梯度缺乏了解。拟议的研究提出了推进这一领域知识状态的各种方法，包括理论研究、实验工作和数据建模的良好融合。

ABSTRACTThis grant is awarded through the Separations and Purification program sub-element of the Interfacial, Transport and Separations program of the Chemical and Transport Systems Division. The principal investigator is Dr. Douglas Frey at the University of Maryland Baltimore County. The proposed research concerns the study of techniques to efficiently purify proteins without exposing them to pH ranges where they might denature or precipitate. These methods are often referred to as buffer focusing chromatography or chromatofocusing. They are not currently used on a large scale because of a lack of understanding amongst practitioners about how retained pH gradients, having a high buffering capacity, can be formed at low ionic strenghts using low cost buffering species. The proposed research suggests various approaches for advancing the state of knowledge in this area and encompasses a good blend of theoretical studies, experimental work and modeling of the data.