Bispecific Monoclonal Antibodies Produced by Somatic Cell Fusion Increase the Potency of Tissue Plasminogen Activator-猫眼课题宝

体细胞融合产生的双特异性单克隆抗体可提高组织纤溶酶原激活剂的效力

DOI：

10.1055/s-0038-1647297

发表时间：

1990

期刊：

Thrombosis and Haemostasis

影响因子：

6.7

通讯作者：

中科院分区：

医学2区

文献类型：

作者：

E. Branscomb;M. Runge;C. Savard;Keith Adams;G. Matsueda;E. Haber

摘要同时与人纤维蛋白和组织型纤溶酶原激活剂(TPA)结合的双特异性单抗可增强tPA的纤溶活性。通过体细胞融合获得了F36.23和F32.1两种双特异性抗体。抗体F36.23的tPA结合来源于抗tPA的单抗TCL8，纤维蛋白的结合来源于抗纤维蛋白的单抗59D8。杂交瘤双特异性抗体F36.23经细胞上清液和腹水两步亲和层析纯化后，在溶液和固相检测中可同时与tPA和纤维蛋白结合。在裂解人纤维蛋白单体的测定中，F36.23将tPA的纤溶效力提高了5~10倍，无论双特异性抗体在测定之前或期间是否与tPA结合。双特异性F36.23F(ab‘)2也同时与tPA和纤维蛋白结合，F36.23F(ab’)2对纤溶的促进作用与完整F36.23的作用相同。第二个双功能抗体F32.1是通过另一种策略产生的，该策略在其他系统中具有更广泛的应用潜力。杂交瘤双特异性抗体F32.1是由免疫脾细胞(用代表人纤维蛋白α链氨基末端的合成寡肽免疫的小鼠)与抗tPA细胞株TCL8融合而成。在固相结合试验和纤溶试验中，杂交瘤双特异性抗体F32.1及其F(ab‘)2与杂交瘤双特异性抗体F36.23的性质无明显差别。体细胞融合产生的双特异性抗体，特别是F(ab‘)2的形式，可能在临床溶栓治疗中具有潜在的应用价值。

Summary Bispecific monoclonal antibodies that bind simultaneously to human fibrin and tissue plasminogen activator (tPA) enhance the fibrinolytic potency of tPA. Two bispecific antibodies (F36.23 and F32.1) were generated by somatic cell fusion. Antibody F36.23 derives its tPA binding from monoclonal anti-tPA antibody TCL8 and its fibrin binding from monoclonal antifibrin antibody 59D8. After purification from cell supernatants and ascites by two steps of affinity chromatography, hybrid-hybridoma bispecific antibody F36.23 simultaneously bound tPA and fibrin in solution and in solid-phase assays. In an assay for the lysis of human fibrin monomer, F36.23 increased the fibrinolytic potency of tPA by 5 to 10 fold, regardless of whether the bispecific antibody had been combined with the tPA before or during the assay. Bispecific F36.23 F(ab′)2 also bound tPA and fibrin simultaneously, and the enhancement in fibrinolysis in the presence of F36.23 F(ab′)2 was identical to that in the presence of intact F36.23. The second bispecific antibody, F32.1, was produced by an alternative strategy that has a wider potential for applicaton in other systems. Hybridoma bispecific antibody F32.1 was derived from the fusion of immune splenocytes (in mice immunized with a synthetic oligopeptide representing the amino terminus of the α-chain of human fibrin) with the anti-tPA cell line TCL8. The properties of hybridoma bispecific antibody F32.1 and its F(ab′)2 were indistinguishable from those of hybrid-hybridoma bispecific antibody F36.23 in solid-phase binding assays and in assays of fibrinolysis. Bispecific antibodies produced by somatic cell fusion, particularly in the form of F(ab′)2, may have potential for use in clinical thrombolysis.