Stabilized Biomimetic Separation Media

描述(申请人提供)：我们将利用聚合物磷脂膜来制备新一代生化分离基质，这些基质在选择性、稳定性、重复性和延展性方面都有显著的改善。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检测器的设计和实现。为了实现这些目标，二氧化硅颗粒(直径从0.5到5微米)和/或二氧化硅毛细管的内壁将被涂上高度稳定的、生物功能化的磷脂双层(PLB)，以便于高度选择性地识别存在于复杂混合物中的跨膜蛋白调节剂。离子通道功能化的PLB将被制备在嵌入在微制造芯片的流道中的微孔上，以从复杂的溶液中识别离子通道调制器。PLB作为基质，通过不同的磷脂头基和/或掺入膜相关和跨膜的蛋白质和受体，使二氧化硅基质具有化学和生物功能。通过形成聚合物支架赋予PLB涂层高度的物理和化学稳定性，将显著提高稳定的PLB固定相的长期稳定性，从而提高其适用性。在稳定的PLB固定相中掺入膜蛋白将为分离和鉴定具有重要生理和药理意义的分析物提供新的基础。除了分离，功能化的PLBS还可以用于新型配体的生物物理分析，例如基于多肽的药物，以确定膜的重要结构特征。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、药物靶标的鉴定和验证、生化途径和新的结合作用的阐明等许多方面。 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，从而能够识别生物功能的新的药理和生理调节因素。这项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化和生物物理参数，以设计下一代基于多肽的药物，用于治疗一系列疾病状态。

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.