组织型纤溶酶原激活物单克隆抗体研制及其特性鉴定

The production and charactor identification of monoclonal antibodies against tissue type plasminogen activator

目的 :应用t PA单克隆抗体建立ELISA法检测t PA含量以及研究t PA功能和结构的关系。方法 :运用杂交瘤技术研制 5株t PA单克隆抗体 ,进行较系统的免疫特性的鉴定。结果 :5株单抗特异性高 ,与u PA、PLG、Fg、Fb、BSA均无交叉反应 ;亲和力强 ,1H4>3C10 >5H10 >4E6 >4C6 ;腹水效价 5× 10 -6～ 1× 10 -7;免疫球蛋白亚类为IgG1和IgG2a ;5株抗体中 ,3C10和 1H4可明显抑制t PA活性 ,而 5H10、4E6、4C6则对t PA活性无明显影响。结论 :为进一步应用这些单抗作为研究手段提供了基础。

Objective:To establish sandwich ELISA assay by self made monoclonal antibody against tissue type plasminogen activator (t PA) and study the relationship between struction and function for t PA.Methods:Successfully produced five hybridoma cell lines of tissue type plasminogen activator and performed the identification of immunologic charactors to great extent.Results:Five monoclonal antibodies have mainly possessed charactors as following.High specificity with no cross reaction with u PA、PLG、Fg、Fb、BSA;strong affinity which showed 1H4>3C10>5H10>4E6>4C6;high titer degree of ascites fluid that was 5×10 -6 ～1×10 -7 and immunoglobulin subclass was IgG1 and IgG2a.Furthermore,among these t PA McAb,3C10、1H4 could obviously inhibit t PA activity while 5H10、4E4、4C6 had no significantly influence on t PA activity.Conclusion:It is suggested that these t PA McAb can almost likely be applied for further study.