Stabilized Biomimetic Separation Media

描述（由申请人提供）：我们将利用聚合磷脂膜制备新一代生化分离基质，其在选择性、稳定性、再现性和可维持性方面表现出显著的改善。我们将集中在两个主要目标：设计和实施a）膜蛋白功能化固定相材料和B）离子通道功能化检测器的微芯片分离。为了实现这些目标，二氧化硅颗粒（直径范围为0.5 μ m至5 μ m）和/或二氧化硅毛细管的内壁将涂覆有高度稳定的生物官能化磷脂双层（PLB），以促进高度选择性地鉴定复杂混合物中存在的跨膜蛋白调节剂。离子通道功能化PLB将在嵌入微加工芯片的流动通道中的微加工孔上制备，以从复杂溶液中识别离子通道调节剂。PLB作为基质，通过不同的磷脂头基和/或结合膜相关和跨膜蛋白和受体，使二氧化硅基质化学和生物功能化。通过形成聚合物支架赋予的PLB涂层的高度物理和化学稳定性将显著改善长期稳定性，从而改善稳定化PLB固定相的适用性。将膜蛋白引入到稳定的PLB固定相中将为分离和鉴定生理和非生理重要分析物提供新的基础。除了分离之外，功能化PLB可用于新型配体（例如基于肽的药物）的生物物理分析，以鉴定膜的结构重要特征。本文提出的研究目标的成功实现将证明对于组合文库的定性和定量分析、药物靶点的鉴定和验证、生化途径和新型结合相互作用的阐明以及许多其他方面是有用的。 公共卫生相关性：该研究项目将开发和实施新的，最先进的生化分离技术，允许识别生物功能的新型药理和生理调节剂。该研究项目还将提供关键的使能技术，从而更有效地阐明关键的生物化学和生物物理参数，用于设计下一代基于肽的药物，用于治疗一系列疾病。

DESCRIPTION (provided by applicant): We will utilize polymeric phospholipid membranes to prepare a new generation of biochemical separation matrices that exhibit marked improvements in selectivity, stability, reproducibility and tenability. We will focus on two primary objectives: design and implementation of a) membrane protein functionalized stationary phase materials and b) ion channel functionalized detectors for microchip separations. To achieve these goals, silica particles (ranging in diameter from 0.5 to 5 <m) and/or the interior walls of silica capillaries will be coated with highly- stabilized, biologically-functionalized phospholipid bilayers (PLB) to facilitate highly selective identification of transmembrane protein modulators existing in complex mixtures. Ion channel functionalized PLBs will be prepared on a microfabricated aperture embedded in the flow channels of a microfabricated chip to identify ion channel modulators from complex solutions. The PLB serves as the matrix for chemically and biologically functionalizing the silica matrices via diverse phospholipid headgroups and/or incorporation of membrane-associated and membrane-spanning proteins and receptors. The high degree of physical and chemical stability of the PLB coatings, imparted via formation of a polymer scaffold, will significantly improve the long-term stability and thereby applicability of stabilized PLB stationary phases. Incorporation of membrane proteins into the stabilized PLB stationary phases will provide a novel basis for separation and identification of physiologically and pharmacologically important analytes. In addition to separations, functionalized PLBs can be used for biophysical analysis of novel ligands, e.g. peptide based pharmaceuticals to identify structurally important features of the membrane. Successful realization of the research goals presented herein will prove useful for qualitative and quantitative analysis of combinatorial libraries, identification and verification of pharmaceutical targets, elucidation of biochemical pathways and novel binding interactions, and many others. PUBLIC HEALTH RELEVANCE: This research project will develop and implement new, state-of-the-art technologies for biochemical separations, allowing for identification of novel pharmacological and physiological regulators of biological function. This research project will also provide key enabling technologies that lead to a more efficient elucidation of key biochemical and biophysical parameters for the design of next generation peptide-based drugs for treatment of a range of disease states.