交叉双单链抗体15C5/c7E3导向的尿激酶原溶栓研究

项目名称： 交叉双单链抗体15C5/c7E3导向的尿激酶原溶栓研究

项目编号： No.30872525

项目类型： 面上项目

立项/批准年度： 2009

项目学科： 轻工业、手工业

项目作者： 谭最

作者单位： 武汉大学

项目金额： 30万元

中文摘要： 血栓形成是临床上常见的疾病，理想的治疗是使用有靶向性的准确而快速的人源化的溶栓剂，本项研究的目的在于合成具有上述功能的溶栓药物。通过对Tomlinson scFv噬菌体文库进行三轮淘洗，成功富集了特异性的38个抗D-dimer抗体和38个抗GPⅡ/Ⅲ抗体；经酶联免疫检测和双脱氧终止法基因测序，获取了高特异性和亲和力的单克隆抗体和其基因序列，并进行了活性和特征的检测鉴定。分别经PCR和双酶切两种连接方法，构建了抗D-Dimer和抗GPIIb/IIIa单链双特异性抗体的质粒并进行表达，与尿激酶原进行偶联，已取得进展，基本完成预定计划，资料正在整理中。在实验中，发现一种更好的回收提取琼脂糖电泳后的DNA特定片段的方法，即陷阱法。

中文关键词： 抗D-dimer抗体；抗GPⅡ/Ⅲ抗体；噬菌体展示技术；单链双特异性抗体；DNA回收

英文摘要： Thrombosis is a common disease and the ideal treatment of thrombosis is to take the drug that is more precise targeting and humanized thrombolytic agent.To achieve such purpose,development of the above thrombolytic agents is important.After three rounds of selection from human scFv phage libraries Tomlinson I and J, 38 monclonal specific anti-D-dimer and 38 anti-GPⅡ/Ⅲ antibodies were picked out.By polyclonal and monoclonal phage ELISA and genes sequencing,full-length monoclonal anti-D-dimer and anti-GPⅡ/Ⅲ antibodies with high specificity and affinity were selected. An anti-D-dimer/anti-GPⅡ/Ⅲ BsAb was constructed by PCR and molecular biological technique of DNA cloning.The fusion protein was expressed and coupled with prourokinase.The research plan was almost finished.In the research,another good method to recover DNA from agarose gel,named trap method,was used.

英文关键词： Anti-D-dimer antibody;Anti-GPⅡ/Ⅲ antibody;Phage display;Bsab;recover DNA