Bio-Optical Composites for Rapid Analyte Detection

描述：(申请人摘要)利用成熟的免疫学和噬菌体展示技术，可以设计出结合几乎任何 具有极强的特异性和亲和力的任意分析物。这些特征表明 生物材料将非常适合用于传感器应用。 然而，不幸的是，没有方便和通用的检测手段 近乎实时的蛋白质-配体结合。潜在的可推广的解决方案 造成这种困难的原因是观察到许多蛋白质只在 结合他们的目标配体。这一折叠实际上代表了最大的 可能发生的物理属性(结构和尺寸)的变化 多肽链。在这里，我们建议合理地引入绑定诱导 折叠成折叠得很好的蛋白质，然后将这么大的， 结合特定的构象变化与容易检测到的 共价连接的光学报告基团的性质。 为了产生特定于结合的光信号，我们将使用共轭 具有优异光学性能的聚合物和半导体材料 那些自然产生的生色团。这些材料起到“传感器”的作用 有能力终止以下物质的光发射的集合 在单个特定的猝灭剂分子存在的情况下的多个物理位置。 通过构建猝灭器-蛋白质-SE结构，我们将产生大量的， 折叠移除猝灭剂时传感器发射率的结合特定变化 从附近到东南部。这为生物传感器提供了手段--光学 具有极大光学放大能力的平台。建议数 研究整合了不同领域的研究人员的免费专业知识 生物化学、有机和无机材料科学和光学领域 光谱学。该提案详细说明了一种适合理性人的方法 结合诱导折叠的产生及共轭化合物的合成 聚合物-蛋白质和无机纳米材料-蛋白质复合材料。还包括 是用于确定这些生物复合材料的效用的简单诊断测试 用于检测重要底物，如特定DNA序列和 逆转录病毒产品。成功完成拟议的研究将证明 一种新的生物光子传感器技术的可行性研究 实时检测重要的临床、工业或 国防利益。

DESCRIPTION: (Applicant's abstract) Using well established immunological and phage-display technologies proteins can be designed that bind almost any arbitrary analyte with great specificity and affinity. These features suggests that biomaterials would be ideally suited for use in sensor applications. Unfortunately, however, there are no convenient and general means of detecting protein-ligand binding in near real-time. A potentially generalizable solution to this difficulty stems from the observation that many proteins fold only upon binding their target ligands. This folding represents effectively the largest possible change in the physical properties (structure and dimensions) of the polypeptide chain. Here we propose to rationally introduce binding-induced folding into otherwise well folded proteins and to couple this large, binding-specific conformational change with an easily detectable change in the properties of covalently attached, optical reporter groups. In order to generate binding-specific optical signals we will use conjugated polymers and semiconductoi ianomaterials with optical properties far superior to those of naturally occurring chromaphores. These materials act as a "sensor ensemble" (SE) that has the capability to terminate the optical emission of multiple )ptical sites in the presence of a single specific quencher molecule. By building a quencher-protein-SE construct, we will generate large, binding-specific changes in sensor emissivity as folding removes the quencher from proximity to the SE. This provides the means for a biosensor-optical platform capable of extremely large optical amplification. The proposed research integrates the complimentary expertise researchers in the diverse fields of biochemistry, organic and inorganic materials science and optical spectroscopy. The proposal details an approach suitable for the rational production of binding-induced folding Lnd the synthesis of conjugated polymer-protein and inorganic nanomaterial-protein composites. Also included are simple diagnostic tests for determining the utility of these biocomposites for the detection of important substrates such as specific DNA sequences and retroviral products. Successful completion of the proposed research will prove the feasibility a novel biophotonic sensor technology suitable for the real-time detection array of compounds of significant clinical, industrial or defense interest.