抗人交联纤维蛋白碎片D-二聚体McAb的制备及其鉴定

Preparation and Characterization of McAbs Against the Human Crosslinked Fibrin Degradation Product D-dimer

以纯化的纤维蛋白碎片D-二聚体为抗原,得到4株分泌抗D-二聚体McAb的杂交瘤细胞,分别命名为HID1、HID2、HID3和HID4,其中HID1、HID2及HID3能识别同一或相邻抗原决定簇,与纤维蛋白原及其碎片X、Y、D、E和血浆等无交叉反应;HID4则识别另一抗原决定簇,且与纤维蛋白原碎片X、Y、D等有交叉反应。本组McAb的成功制备,将为临床血栓性疾病的诊断和治疗监测提供新手段。

D-dimer is a specific degradation product of crosslinked fibrin hydrolyzed by plasmin.Detection of D-dimer in blood is important for the diagnosis of throm-botic diseases and the surveillance of thromobolytic treatment. Therefore,D-dimer purification and preparation of anti-D-dimer McAb are necessary tasks. Using routine methods, we prepared four hybridomas which secrete anti-D-dimer McAb: HID1,HID2,HID3 and HID4. After five months in culture these hybridomas still secrete anti-D-dimer McAb. The titer of ascites McAb is 1:10-5-1:10-6. HID1, HID2 and HID3 recognize the same or similar antigenic determinants and don't cross-react with coagulation factors, plasma or fibri-nogen degradation products X, Y, D, and E fragments. HID4 recognizes another antigenic determinant and cross-reacts with fibrinogen degradation products X, Y and D fragments. Immunoblot results show that all of these McAbs react with a MW 190 000 band.These antibodies are being used in the development of enzyme-linked and latex immunoassay methods for the detection of D-dimer and will be useful in the diagnosis, evaluation and monitoring of thrombotic diseases and thrombolytic treatment.