Solid-phase assays for biosensing and high throughput screening using sol-gel derived bioglasses

这项研究的主要目标是：(1)开发生物相容的糖或多元醇修饰的溶胶-凝胶前驱体和新的多肽功能化双硅烷，它们将在二氧化硅材料中提供可调的表面性质和可控的形貌，并为被包裹的生物分子提供良好的长期稳定性；(2)利用先进的荧光(时间分辨各向异性，FRAP)和物理表征方法(扫描电子显微镜、原子力显微镜、孔压计)表征由这些前驱体衍生的新材料；(3)检测各种内在荧光和外在标记的蛋白质和适配子的热力学稳定性、可及性、构象、动力学和长期稳定性，以全面表征蛋白质功能与溶胶-凝胶材料特性和老化之间的相关性；(4)扩展我们基于溶胶-凝胶的微阵列平台，包括基于膜结合蛋白质和DNA适配子的新应用，用于纳米体积高温超导和多分析生物传感；(5)进一步开发我们基于溶胶-凝胶的生物亲和层析和酶反应器柱及方法，集成多维LC和MALDI MS/MS平台，用于小分子筛选和诊断以及使用蛋白质或适配子进行生物定向固相萃取。一个主要目标将是将这些检测技术应用于重要的生物靶点，特别是可以对健康研究产生重大影响的激酶和GPCRs。

The major objectives of the proposed research are: (1) the development of biocompatible sugar or polyol modified sol-gel precursors and new peptide functionalized bis-silanes that will provide tunable surface properties and controlled morphologies in silica materials along with good long-term stability for entrapped biomolecules; (2) the characterization of new materials that are derived from these precursors using advanced fluorescence (time resolved anisotropy, FRAP) and physical characterization methods (SEM, AFM, porosimetry); (3) the examination of the thermodynamic stability, accessibility, conformation, dynamics and long-term stabiity of a variety of intrinsically fluorescent and extrinsically labelled proteins and aptamers to comprehensively characterize correlations between protein function and sol-gel material properties and aging; (4) expansion of our sol-gel based microarray platform to include new applications based on membrane-bound proteins and DNA aptamers for nanovolume HTS and multianalyte biosensing; (5) further development of our sol-gel based bioaffinity chromatography and enzyme reactor columns and assays to integrate multidimensional LC and MALDI MS/MS platforms for small molecule screening and diagnostics and for biologically directed solid phase extraction using either proteins or aptamers. A major goal will be to apply these assay technologies to important biological targets, particularly kinases and GPCRs, where a significant impact on health research can be made.