THROMBUS DETECTION USING FIBRIN-SPECIFIC ANTIBODIES

使用纤维蛋白特异性抗体检测血栓

• 批准号: 3339431
• 负责人: GARY R MATSUEDA
• 金额: $ 15.06万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1982
• 资助国家: 美国
• 起止时间: 1982-01-01 至 1987-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3339431
• 关键词: antibody specificity; cardiovascular disorder diagnosis; chemical cleavage; chromatography; crosslink; early diagnosis; fibrin; fibrinogen; hybridomas; immunochemistry; immunoconjugates; immunohematology; immunologic techniques; monoclonal antibody; noninvasive diagnosis; plasmin; radionuclide diagnosis; thrombosis

Thrombus detection remains a challenge to the physician who must manage certain life-threatening problems such as coronary artery thrombi or pulmonary emboli neither of which can be reliably diagnosed. The long term objective of the proposed research is the use of fibrin specific antibodies to localize intravascular deposits of fibrin, a constitutive protein network in thrombi. During the current grant period, a novel immunochemical strategy to obtain fibrin specific antibodies has been validated: a synthetic peptide, chosen to mimic the amino terminus of the Beta-chain in human fibrin, has been used to derive three monoclonal anti(human)fibrins, 64C5, 55D10, and 59D8. The aims of the present grant proposal (Y04-Y06) are to 1) develop suitable animal models for a quantitative evaluation of Mab-64C5, -55D10 and -59D8 as thrombus detecting agents and 2) prepare different antifibrin antibodies as alternative thrombus detecting agents. Both chicken and rabbits will be used to develop models suitable for detection of experimentally induced thrombi using radiolabeled monoclonal antifibrins. If the results are promising, considerations unique to in vivo scintigraphic imaging will be investigated; ie. choice of ideal radioisotope (eg. 99mTc or 111In), non-denaturing labelling with radioisotope, and biodistribution studies. As a part of this objective, we intend to characterize the manner in which 64C5, 55D10, and 59D8 recognize fibrin ad its circulating degradation products. The likehood of immunocrossreactivity with degradation products and circulating fibrin monomers is the basis for the second objective, the search for alternative antifibrin antibodies. We will synthesize fibrin-unique peptides for us as antigens: the amino terminal heptapeptide of the Alpha-chain in human fibrin, the Factor XIIIa mediated crosslink site in Gamma-chain dimers (2 peptides), and the amino terminal heptapeptides exposed upon plasmic cleavage of fibrin Beta-chains. These peptides will then be used as antigens to immunize rabbits and mice with the aim of obtaining antibodies which bind to human fibrin but not fibrinogen.

血栓检测仍然是一个挑战，医生谁必须管理 某些危及生命的问题，如冠状动脉血栓或 肺栓塞，两者都不能可靠地诊断。 长期 本研究的目的是利用纤维蛋白特异性抗体 定位血管内纤维蛋白沉积物， 血栓网 在目前的资助期内，一部小说 获得纤维蛋白特异性抗体免疫化学策略 验证：合成肽，选择模拟的氨基末端的 人纤维蛋白中的β-链，已被用于衍生三种单克隆抗体， 抗（人）白蛋白、64 C5、55 D10和59 D8。 本补助金的目的 建议（Y 04-Y 06）是1）开发合适的动物模型， 单克隆抗体-64C5、-55D10和-59D8作为血栓检测的定量评价 试剂和2）制备不同的抗纤维蛋白抗体作为替代 血栓检测剂。 鸡和兔子都将被用来 开发适用于检测实验诱导血栓的模型 使用放射性标记的单克隆抗人胰岛素。 如果结果是有希望的， 体内造影成像的独特考虑因素将是 调查;即。选择理想的放射性同位素（例如，99 mTc或111 In）， 放射性同位素非变性标记和生物分布研究。 作为这一目标的一部分，我们打算描述 64 C5、55 D10和59 D8识别纤维蛋白及其循环降解 产品. 与降解产物的免疫交叉反应性相似性 和循环纤维蛋白单体是第二个目标的基础， 寻找替代的抗纤维蛋白抗体。 我们将合成 纤维蛋白特异性抗原肽：氨基端七肽 人纤维蛋白中α-链，因子XIIIa介导的交联 γ-链二聚体（2个肽）中的位点和氨基末端 在纤维蛋白β-链的胞质裂解时暴露的七肽。 这些 然后将肽用作抗原以免疫兔子和小鼠， 获得结合人纤维蛋白但不 纤维蛋白原。