SBIR PHASE I: Metal Cluster and Combined Flourescent/Gold Labels for Peptide Synthesis

SBIR 阶段 I：用于肽合成的金属簇和组合荧光/金标记

• 批准号: 9561345
• 负责人: Richard Powell
• 金额: $ 7.5万
• 依托单位: Nanoprobes Incorporated
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-03-01 至 1996-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9561345&HistoricalAwards=false
• 关键词: SBIR; PHASE; Metal; Cluster; Combined

9561345 Powell In this Small Business Innovation Research Phase I project, Undecagold, 1.4 nm "Nanogold" cluster complexes, combined fluorescent and gold cluster labels, and platinum cluster complexes 1.8 to 3.6 nm in diameter will be covalently conjugated to protected amino acids suitable for supported peptide synthesis. Analogs of L-cysteine, L-lysine and glycine will be prepared, in addition to labeling reagents functionalized to append the cluster labels to the N-terminal and C-terminal, and will be used to insert the labels at programmed positions in synthetic peptides during automated peptide synthesis. Chirality of the natural amino acids will be preserved to retain maximum peptide bioactivity. The labels will be synthetically modified for maximum solubility and stability towards synthesis and deprotection conditions by systematic variations of their coordinated ligands. Labeled peptide analogs of the C-terminal region of histone H1 protein, HMG-D and HMG-Z proteins will be prepared and microinjected into living cells. Their distribution, function and macromolecular localization within the cell nucleus will be studied by fluorescence confocal and electron microscopy. Labels will also be incorporated into synthetic analogs of the hormone peptide leuteinizing hormone releasing hormone (LHRH), then their sensitivity and specificity as immunoassay probes will be studied in membrane blot and microtiter plate tests. The reagents will be widely used by electron microscopists, cell and molecular biologists to prepare probes with the highest possible resolution, retention of bioactivity and sensitivity. The strategy for preparing labeled and unlabeled peptides simultaneously will reduce probe preparation time and cost. Rapidly growing applications exist in the selective labeling of combinatorial peptides. These reagents may also be used to prepare novel hybrid materials in which the unique electronic and catalytic properties of metal nanoparticles are programmed site-specif ically into new biomaterials to yield new macromolecular devices.

9561345 Powell 在这个小型企业创新研究第一阶段项目中，Undecagold、1.4 nm“Nanogold”簇复合物、组合荧光和金簇标记以及直径 1.8 至 3.6 nm 的铂簇复合物将共价缀合到适合支持肽合成的受保护氨基酸。 除了功能化以将簇标记附加到 N 端和 C 端的标记试剂之外，还将制备 L-半胱氨酸、L-赖氨酸和甘氨酸的类似物，并将用于在自动肽合成过程中将标记插入到合成肽中的编程位置。 天然氨基酸的手性将被保留，以保留最大的肽生物活性。 通过对其配位体的系统变化，对标记进行合成修饰，以获得最大的溶解度和对合成和脱保护条件的稳定性。 组蛋白 H1 蛋白、HMG-D 和 HMG-Z 蛋白 C 端区域的标记肽类似物将被制备并显微注射到活细胞中。 将通过荧光共聚焦和电子显微镜研究它们在细胞核内的分布、功能和大分子定位。 标签还将被纳入激素肽黄体生成素释放激素（LHRH）的合成类似物中，然后它们作为免疫测定探针的敏感性和特异性将在膜印迹和微量滴定板测试中进行研究。 这些试剂将被电子显微镜学家、细胞和分子生物学家广泛使用，以制备具有尽可能高的分辨率、保留生物活性和灵敏度的探针。 同时制备标记和未标记肽的策略将减少探针制备时间和成本。 组合肽选择性标记的应用正在快速增长。 这些试剂还可用于制备新型混合材料，其中金属纳米颗粒独特的电子和催化特性被编程到新的生物材料中，以产生新的大分子器件。