NOVEL BINDING MOLECULES FOR OPIOID PEPTIDES

在这个项目中进行的研究将提供具体的约束力 用于阿片肽分子的定量，使用一种创新的 多肽结合体的合成、选择和分析方案 结构。在第一阶段，将通过以下方法合成结合分子 一种新的模板技术。这项技术涉及将 靶标多肽为固体载体并在其周围构建聚合物 具有多个特定的非共价点交互的网络 多肽。有希望的结合结构将被标记为 荧光团，并将通过将多肽偶联到 光纤和执行竞争性结合分析使用 荧光标记的活页夹。固态核磁共振分析将表明 选定结构的均质程度，当与 使用13C标记的模板肽，将提示识别基团 专门与目标功能团体在 多肽。在第二阶段，我们将细化模板方案，并使用 来自固态核磁共振的结构数据用于插入共识结构 它可以通过溶液化学来制备，不需要 扩展中的模板。我们还将准备可识别的活页夹 额外的阿片肽，包括肽前体，并使用这些 测定粘合剂的加工活性、酶活性。 建议的商业应用： 这些合成粘结剂可以提供常规粘结剂无法提供的特性 抗体和受体，包括增强对酶的稳定性 水解，缩小尺寸，不需要动物或组织培养的生产， 易于扩大制造规模并由此降低成本， 无免疫原性，在非水溶剂中使用的潜力。这些 分子可以适应任何检测形式，并有可能 用于分析和制备分离，以及用于治疗 申请。

The research performed in this project will provide specific binding molecules for the quantitation of opioid peptides, using an innovative scheme for synthesis, selection and analysis of peptide-binding structures. In Phase I, binding molecules will be synthesized by means of a novel templating technology. This technology involves coupling the target peptide to a solid support and building around it a polymer network that has many specific, noncovalent points of interaction with the peptide. Promising binding structures will be labeled with a fluorophore, and feasibility will be proven by coupling peptides to optical fibers and performing competitive binding assays using fluorescent-labeled binders. Solid state NMR analysis will indicate the degree of homogeneity of selected structures and, when coupled with the use of 13C-labeled template peptide, will suggest recognition groups that specifically interact with targeted functional groups on the peptide. In Phase II we will refine templating schemes, and use structural data from solid state NMR to interpolate consensus structures that can be prepared by solution chemistry, eliminating the need for templates in scaling up. We also will prepare binders that recognize additional opioid peptides, including peptide precursors, and use these binders to determine the activity of processing, enzymes. PROPOSED COMMERCIAL APPLICATION: These synthetic binders can provide features unavailable in conventional antibodies and receptors, including enhanced stability to enzymatic hydrolysis, reduced size, production without animals or tissue culture, ease of manufacturing scale-up and the resulting reduction in cost, nonimmunogenicity, and potential for use in non-aqueous solvents. These molecules can be adapted to any assay format, and have the potential for use in analytical and preparative separations, and for therapeutic applications.