Enhanced Protein Separations Using Molecular Micelles

描述(由申请者提供)：虽然基于前一次美国国立卫生研究院拨款期间进行的研究，但这项提案概述了关注根本不同方向的研究。这项提案中概述的研究的总体目标是探索分离小蛋白质样本的改进方法。为实现这一总体目标，概述了三个重点领域。第一个重点领域是主要的研究领域，包括开发用于分离微升蛋白质的开管毛细管电色谱(OT-CEC)。在第一个领域，通过交替清洗阳离子和阴离子聚合物产生的非常坚固的聚电解质多层(PEM)涂层将用于OT-CEC模式。我们研究的这一阶段最初将使用的离子聚合物将以分子胶束为基础，类似于我们在上一个资助期开发的那些分子胶束。随着研究的发展，还将添加其他阴离子聚合物。阳离子聚合物也将是类似于我们上一个资助期使用的聚合物(例如聚赖氨酸)。然而，新的阳离子分子胶束也将被合成，并与我们的阴离子分子胶束结合作为PEM涂层的新形式进行探索。此外，我们还将探索两性聚合物在OT-CEC中用于PEM涂层的用途。这项研究的一个关键组成部分是了解我们的PEM涂层的结构。我们的PEM涂层将与麻省理工学院化学工程系的Paula Hammond博士合作进行表征。这项拟议研究的第二个重点领域是探索在第一阶段开发的OT-CEC技术在微流控平台上的适用性，以分析纳升体积的蛋白质。这项研究将与美国国家标准与技术研究所的劳里·洛卡西奥博士合作完成。这项研究的第三阶段涉及使用分子胶束来提高凝胶电泳法分离蛋白质的能力。与其他重点领域一样，将利用各种策略来优化不同类别蛋白质的分离。最后，需要指出的是，由于蛋白质无处不在，该项目与生物医学研究的几乎所有领域都有关。因此，分离、检测和鉴定这一提案中概述的少量蛋白质对公共卫生的几乎所有方面都很重要。

DESCRIPTION (provided by applicant): While based on studies performed during the previous NIH grant period, this proposal outlines research which focuses in a fundamentally different direction. The overall goal of the research outlined in this proposal is to explore enhanced methods for separation of small protein samples. Three focus areas are outlined to achieve this overall goal. The first focus area is the primary area of research and involves development of open-tubular capillary electrochromatography (OT-CEC) for separation of microliter quantities of proteins. In this first area, very robust polyelectrolyte multilayer (PEM) coatings generated from alternate rinses of cationic and anionic polymers will be used in the OT-CEC mode. The ionic polymers which will be initially used in this phase of our research will be based on molecular micelles similar to those developed during our previous funding period. Other anionic polymers will be added as the research develops. The cationic polymers will also be polymers (e.g. poly-lysine) similar to those used during our previous funding period. However, new cationic molecular micelles will also be synthesized and explored in combination with our anionic molecular micelles as new forms of PEM coatings. In addition, we will explore the utility of zwitterionic polymers for PEM coatings in OT-CEC. A critical component of this research is to understand the structure of our PEM coatings. Characterization of our PEM coatings will be done in collaboration with Dr. Paula Hammond of the department of chemical engineering of MIT. The second focus area of this proposed research is to explore the applicability of the OT-CEC technology developed during the first phase to microfluidic platforms for analyses of nanoliter volumes of proteins. This research will be done in collaboration with Dr. Laurie Locascio of the National Institute of Standards and Technology. The third phase of this research involves the use of molecular micelles to improve the separation of proteins using gel electrophoresis. As with the other focus areas, various strategies will be exploited to optimize separations for different classes of proteins. Finally, it is noted that this project is relevant to almost all areas of biomedical research since proteins are ubiquitous. Thus, isolation, detection, and identification of small quantities of proteins as outlined in this proposal are important to almost all aspects of public health.