THROMBUS DETECTION USING FIBRIN-SPECIFIC ANTIBODIES

使用纤维蛋白特异性抗体检测血栓

• 批准号: 3339430
• 负责人: GARY R MATSUEDA
• 金额: $ 15.57万
• 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/3339430
• 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.

血栓检测对医生来说仍然是一项挑战，他们必须 某些危及生命的问题，如冠状动脉血栓或 这两种情况都不能得到可靠的诊断。从长远来看 拟议研究的目的是使用纤维蛋白特异性抗体 定位血管内纤维蛋白沉积，纤维蛋白是一种构成蛋白 血栓形成的网状结构。在目前的资助期内，一部小说 获得纤维蛋白特异性抗体的免疫化学策略已经被 有效：一种合成肽，选择来模拟 人纤维蛋白中的β链，已经被用来衍生出三种单抗 抗(人)纤维蛋白、64C5、55D10和59D8。这笔赠款的目的 建议(Y04-Y06)是1)开发适合的动物模型 单抗-64C5、-55D10和-59D8检测血栓的定量评价 试剂和2)制备不同的抗纤维蛋白抗体作为替代 血栓检测试剂。鸡和兔子都将被用来 开发适合于检测实验诱导血栓的模型 使用放射性标记的单抗纤维蛋白。如果结果令人振奋， 活体核素成像的独特考虑因素将是 已调查；即理想放射性同位素的选择(例如。99mTc或111In)， 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 64C5、55D10和59D8识别纤维蛋白及其循环降解 产品。免疫交叉反应与降解产物的亲和性 循环中的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。我们将综合 纤维蛋白作为抗原的独特多肽：氨基末端七肽 人纤维蛋白α链的XIIIA因子介导的交联物 伽马链二聚体(2个肽)中的位点和氨基末端 暴露在纤维蛋白β链胞质裂解上的七肽。这些 然后用多肽作为抗原免疫兔子和小鼠 获得与人纤维蛋白结合但不结合的抗体的目的 纤维蛋白原。