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: 6889247
• 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 2007-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6889247
• 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; phage display; 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：（申请人的摘要）使用良好的免疫和噬菌体 - 播放技术蛋白质可以设计几乎所有绑定 具有特异性和亲和力的任意分析物。这些功能暗示 该生物材料非常适合在传感器应用中使用。 但是，不幸的是，没有方便和一般的检测手段 蛋白质 - 配体结合在接近实时。一种可能概括的解决方案 这一困难源于观察到许多蛋白质仅在 绑定其靶配体。此折叠有效地表示最大 可能会改变物理特性（结构和维度） 多肽链。在这里，我们建议合理引入结合诱导的 折叠成原本折叠良好的蛋白质，并将其融合到这么大， 结合特异性构象变化，易于检测到可检测的变化 共价附加的光学报告基团的特性。 为了生成绑定特定的光学信号，我们将使用共轭 具有光学特性的聚合物和半导体材料材料 对于天然发生的染色体。这些材料充当“传感器 合奏”（SE），具有终止光发射的能力 在单个特异性淬灭分子的存在下，多个）ptical位点。 通过构建淬火器 - 蛋白质-SE结构，我们将产生大型， 随着折叠去除淬火器，传感器发射率的结合特异性变化 从接近到SE。这为生物传感器 - 光学提供了手段 能够具有极大的光学扩增的平台。提议 研究将免费的专业知识研究人员融合在一起 生物化学，有机和无机材料科学和光学领域 光谱法。该提案详细介绍了一种适合理性的方法 结合引起的折叠的产生lnd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 是简单的诊断测试，用于确定这些生物复合材料的实用性 用于检测重要底物，例如特定的DNA序列和 逆转录病毒产品。成功完成拟议的研究将证明 可行性是一种新型的生物光传感器技术，适合 大量临床，工业或 国防利益。