Enhanced Protein Separations Using Molecular Micelles

使用分子胶束增强蛋白质分离

• 批准号: 7487946
• 负责人: ISIAH M WARNER
• 金额: $ 25.97万
• 依托单位: LOUISIANA STATE UNIV A&M COL BATON ROUGE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-20 至 2010-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7487946
• 关键词: Area; Arterial Fatty Streak; Arteries; Biological Markers; Biomedical Research; Blood capillaries; Bypass; Charge; Chemical Engineering; Chemistry; Class; Collaborations; Complex Mixtures; Coupled; Data; Detection; Development; Devices; Film; Funding; Glycoproteins; Goals; Grant; Institutes; Laboratories; Lysine; Malignant Neoplasms; Measurement; Methods; Micelles; Microfluidic Microchips; Microfluidics; Molecular; Phase; Plague; Polymers; Procedures; Proteins; Public Health; Reagent; Research; Research Personnel; Sampling; Solutions; Standards of Weights and Measures; Structure; Technology; Tubular formation; Two-Dimensional Gel Electrophoresis; United States National Institutes of Health; Work; base; capillary; concept; desire; gel electrophoresis; improved; interest; mass spectrometer; molecular recognition; nanolitre; nanolitre scale; novel; success; tool

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.

描述（由申请人提供）：虽然基于在以前的NIH资助期间进行的研究，但本提案概述了侧重于根本不同方向的研究。该提案中概述的研究的总体目标是探索用于分离小蛋白样品的增强方法。为实现这一总体目标，概述了三个重点领域。第一个重点领域是研究的主要领域，涉及开发用于分离微升量蛋白质的开管毛细管电色谱（OT-CEC）。在这第一个领域中，将在OT-CEC模式中使用由阳离子和阴离子聚合物的交替冲洗产生的非常坚固的可渗透多层（PEM）涂层。在我们研究的这一阶段中最初使用的离子聚合物将基于与我们之前资助期间开发的分子胶束类似的分子胶束。随着研究的发展，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之前资助期间使用的聚合物（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，我们将探索两性离子聚合物在OT-CEC中用于PEM涂层的效用。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涂层的表征将与麻省理工学院化学工程系的Paula哈蒙德博士合作完成。这项拟议研究的第二个重点领域是探索第一阶段开发的OT-CEC技术对微流体平台的适用性，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改善凝胶电泳对蛋白质的分离。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，值得注意的是，这个项目是相关的几乎所有领域的生物医学研究，因为蛋白质是无处不在的。因此，分离、检测和鉴定本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。