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

本研究的主要目标是：（1）开发生物相容的糖或多元醇改性的溶胶-凝胶前体和新型肽官能化的双硅烷，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态，同时沿着对包埋的生物分子具有良好的长期稳定性;（2）使用先进的荧光技术对由这些前体衍生的新材料进行表征（时间分辨各向异性，FRAP）和物理表征方法（SEM、AFM、孔隙率测定法）;（3）热力学稳定性、可及性、构象、动态和长期稳定性的各种内在荧光和外源标记的蛋白质和适体，以全面表征蛋白质功能和溶胶之间的相关性，凝胶材料的性质和老化;（4）我们的基于溶胶-凝胶的微阵列平台的扩展，以包括基于膜结合蛋白质和DNA适体的纳米体积HTS和多分析物生物传感的新应用;（5）进一步开发我们的基于溶胶-凝胶的生物亲和色谱和酶反应器柱和测定，以整合多维LC和MALDI MS/MS。用于小分子筛选和诊断以及使用蛋白质或适体进行生物定向固相萃取的MS平台。一个主要目标是将这些检测技术应用于重要的生物靶点，特别是激酶和GPCR，这对健康研究产生了重大影响。

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.