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物化学分离基质，该基质在选择性、稳定性、可重复性和可持续性方面有显著改善。我们将专注于两个主要目标：a)膜蛋白功能化固定相材料的设计和实现；b)用于微芯片分离的离子通道功能化探测器。为了实现这些目标，二氧化硅颗粒（直径范围从0.5到5 <m）和/或二氧化硅毛细血管的内壁将被高度稳定的生物功能化磷脂双分子层（PLB）包裹，以促进在复杂混合物中存在的跨膜蛋白调节剂的高度选择性鉴定。离子通道功能化PLBs将在嵌入微加工芯片流道的微加工孔径上制备，以从复杂溶液中识别离子通道调制器。PLB通过不同的磷脂头基团和/或结合膜相关蛋白和膜跨越蛋白和受体，作为化学和生物功能化二氧化硅基质的基质。通过聚合物支架的形成，PLB涂层具有高度的物理和化学稳定性，这将显著提高稳定PLB固定相的长期稳定性，从而提高其适用性。将膜蛋白掺入稳定的PLB固定相将为分离和鉴定生理和药理学上重要的分析物提供新的基础。除了分离之外，功能化的plb还可以用于新型配体的生物物理分析，例如基于肽的药物，以识别膜的结构重要特征。本文提出的研究目标的成功实现将有助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。

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.