Stabilized Biomimetic Separation Media

稳定的仿生分离介质

• 批准号: 8258754
• 负责人: CRAIG A ASPINWALL
• 金额: $ 25.72万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8258754
• 关键词: Accounting; Adsorption; Affect; Binding; Biochemical; Biochemical Pathway; Biological Assay; Biological Process; Biomimetics; Blood capillaries; Caliber; Cells; Chemicals; Complex; Complex Mixtures; Detection; Development; Disease; Ensure; Exhibits; G-Protein-Coupled Receptors; Generations; Goals; Integral Membrane Protein; Intervention; Ion Channel; Lead; Libraries; Ligands; Lipids; Marketing; Membrane; Membrane Proteins; Methods; Microfluidics; Molecular Bank; Peptides; Performance; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Phospholipids; Physiological; Polymers; Preparation; Property; Proteins; Regulation; Reproducibility; Research; Research Project Grants; Sampling; Screening procedure; Series; Silicon Dioxide; Solutions; Technology; Tubular formation; base; capillary; chemical stability; combinatorial; design; detector; improved; innovation; microchip; next generation; novel; particle; polymerization; protein function; public health relevance; receptor; reconstitution; response; scaffold; surface coating

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.

描述（由申请人提供）：我们将利用聚合磷脂膜来制备新一代生化分离基质，其在选择性、稳定性、重现性和耐用性方面表现出显着的改进。我们将重点关注两个主要目标：a) 膜蛋白功能化固定相材料和 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（直径范围为 0.5 至 5 µm）和/或二氧化硅毛细管的内壁将涂有高度稳定的生物功能化磷脂双层 (PLB)，以促进跨膜的高度选择性识别存在于复杂混合物中的蛋白质调节剂。离子通道功能化 PLB 将在嵌入微加工芯片流道中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 作为基质，通过不同的磷脂头基和/或掺入膜相关蛋白和跨膜蛋白和受体，对二氧化硅基质进行化学和生物功能化。通过形成聚合物支架而赋予 PLB 涂层高度的物理和化学稳定性，将显着提高长期稳定性，从而提高稳定 PLB 固定相的适用性。将膜蛋白掺入稳定的 PLB 固定相将为生理学和药理学重要分析物的分离和鉴定提供新的基础。除了分离之外，功能化 PLB 还可用于新型配体的生物物理分析，例如配体。基于肽的药物来识别膜的重要结构特征。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、生化途径和新颖的结合相互作用的阐明以及许多其他方面是有用的。 公共健康相关性：该研究项目将开发和实施新的、最先进的生化分离技术，从而识别生物功能的新型药理学和生理调节剂。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化和生物物理参数，以设计下一代基于肽的药物来治疗一系列疾病状态。