THROMBUS DETECTION USING FIBRIN-SPECIFIC ANTIBODIES

使用纤维蛋白特异性抗体检测血栓

• 批准号: 3339434
• 负责人: GARY R MATSUEDA
• 金额: $ 24.35万
• 依托单位: PRINCETON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-01-01 至 1993-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3339434
• 关键词: aminoacid analyzer; antibody specificity; antigens; binding proteins; cardiovascular disorder diagnosis; crosslink; diagnosis design /evaluation; diagnosis quality /standard; disease /disorder model; embolism; fibrin; fibrinogen; high performance liquid chromatography; hybridomas; immunization; immunochemistry; immunoconjugates; immunohematology; immunologic techniques; immunotherapy; iodine; laboratory mouse; laboratory rabbit; mass spectrometry; monoclonal antibody; noninvasive diagnosis; peptide analog; peptide chemical synthesis; protein sequence; radionuclide diagnosis; radionuclide imaging /scanning; radionuclides; synthetic peptide; thrombosis

Current methods for in vivo thrombus detection either lack specificity or pose additional health risks. With the tools of molecular immunology and syntheitic peptide chemistry, we plan to prepare a clinically useful, diagnostic antibody that possesses specificity for human fibrin (the structural protein in thrombi) and provides sufficient sensitivity for in vivo thrombus detection by gamma scintigraphy. Our strategy will emphasize the differences between fibrin and its soluble precursor, fibrinogen, and will focus on ways to select fibrinspecific antibodies. Synthetic peptides will be designed to mimic fibrinunique epitopes and used to elicit antibodies that will bind human fibrin in the presence of fibrinogen. This approach was validated during the previous grant period when we used a synthetic peptide corresponding to the N terminus of the human fibrin beta-chain to elicit monoclonal antibodies that bound to fibrin monomer in the presence of fibrinogen. The in vivo specificity of one monoclonal antibody (64C5) was confirmed in experiments using chicken, rabbit and dog models. In the research proposed here, we focus on the design and testing of various fibrin-like peptides corresponding to thrombin cleavage sites and Factor XIIIa cross-link sites, and describe plans for an improved screening strategy that uses fibrin miniclots rather than fibrin monomers as antigens. Our specific aims include the in vivo testing of antibody 5D7 as a gamma scintigraphic agent for detecting experimental thrombi; the preparation of monoclonal antibodies that avidly bind to clotted fibrin; the synthesis and refinement of cross-link antigens related to gamma-chain cross-link sites; and the structural and immunochemical study of the alpha-chain cross-link site in order to identify a new fibrin-unique epitope. Three specific benefits will result from these experiments: First, they will provide a detailed immunochemical analysis of unique epitopes on the fibrin molecule. They will also suggest general methods for differentiating any protein from its precursor. Second, they will provide reagents that can tag thrombi in vivo. Third, they will lay the foundation upon which a sensitive and specific noninvasive test for life-threatening thromboembolism can be built.

目前体内血栓检测的方法要么缺乏 专一性或构成额外的健康风险。借助以下工具 分子免疫学和合成肽化学，我们计划 制备一种临床有用的诊断抗体，它具有 人纤维蛋白(血栓中的结构蛋白)和 为体内血栓检测提供足够的灵敏度 伽马闪烁照相术。我们的战略将强调差异 在纤维蛋白及其可溶性前体纤维蛋白原之间，并将焦点 关于选择纤维蛋白特异性抗体的方法。合成肽 将被设计成模拟纤维蛋白独特的表位并用于诱导 在人纤维蛋白存在的情况下结合人纤维蛋白的抗体 纤维蛋白原。此方法在上一次拨款期间得到了验证 当我们使用与N相对应的合成肽时 人纤维蛋白β链末端诱导的单克隆 在存在的情况下与纤维蛋白单体结合的抗体 纤维蛋白原。一株单抗的体内特异性 (64C5)在用鸡、兔和 狗狗模特。在这里提出的研究中，我们将重点放在设计 和各种纤维蛋白样多肽的检测 凝血酶裂解位点和凝血因子XIIIA交联位点，以及 描述使用纤维蛋白的改进筛查策略的计划 作为抗原的是小颗粒而不是纤维蛋白单体。我们的特定 目的包括体内检测抗体5D7作为一种伽马 用于检测实验性血栓的闪光显像剂； 与凝血蛋白亲和的单抗的制备 纤维蛋白；相关交联性抗原的合成和提纯 到伽马链交联位点；以及结构和 α链有序交联位的免疫化学研究 以确定一种新的纤维蛋白独特的表位。三个具体好处 将从这些实验中产生：首先，它们将提供一个 纤维蛋白独特表位的详细免疫化学分析 分子。他们还将提出一般的方法，以 将任何蛋白质与其前体区分开来。第二，他们会 提供可以标记体内血栓的试剂。第三，它们会 敏感和特定的非侵入性的基础 可以建立危及生命的血栓栓塞症测试。