Bio-Optical Composites for Rapid Analyte Detection

用于快速分析物检测的生物光学复合材料

• 批准号: 6636625
• 负责人: Kevin W Plaxco
• 金额: $ 21.42万
• 依托单位: UNIVERSITY OF CALIFORNIA SANTA BARBARA
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-05-01 至 2006-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6636625
• 关键词: analytical method; bioengineering /biomedical engineering; biosensor device; chemical conjugate; chemical structure function; circular dichroism; conformation; covalent bond; cystine; fluorescence spectrometry; intermolecular interaction; ligands; lysine; nanotechnology; optics; peptide chemical synthesis; polymers; protein binding; protein engineering; protein folding; reagent /indicator; semiconduction; site directed mutagenesis; thermodynamics; time resolved data; transcription factor

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.

描述：（申请人摘要）使用完善的免疫学和 噬菌体展示技术蛋白质可以被设计成结合几乎任何 任意分析物具有很大的特异性和亲和力。这些特征表明 生物材料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， 近实时的蛋白质-配体结合。一个潜在的可推广的解决方案 这一困难源于观察，许多蛋白质折叠只有在 结合它们的靶配体。这种折叠实际上代表了 物理性质（结构和尺寸）的可能变化 多肽链在这里，我们建议合理地引入绑定诱导 折叠成其他折叠良好的蛋白质， 结合特异性构象变化，在结合特异性构象中具有容易检测的变化。 共价连接的光学报告基团的性质。 为了产生结合特异性光信号，我们将使用共辄光信号。 光学性能远优于上级的聚合物和纳米材料 与自然产生的发色团的颜色不同。这些材料充当“传感器 具有终止光发射的能力的“系综”（SE）。 多个）光学位点。 通过构建猝灭蛋白-SE构建体，我们将产生大量， 当折叠去除猝灭剂时，传感器发射率的结合特异性变化 从接近SE。这为生物传感器提供了一种方法， 能够进行极大光放大的平台。拟议 研究整合了互补的专业知识研究人员在不同的 生物化学、有机和无机材料科学和光学领域 谱该提案详细说明了一种适合理性的方法， 结合诱导折叠的产生和共轭 聚合物-蛋白质和无机纳米材料-蛋白质复合物。还包括 是确定这些生物复合材料效用的简单诊断测试 用于检测重要的底物，如特异性DNA序列， 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， 具有重要临床意义的化合物的实时检测阵列， 国防利益。