Enhancement of Clot Lysis In Vitro and In Vivo with a Bispecific Monoclonal Antibody Directed against Human Fibrin and against Urokinase-Type Plasminogen Activator
Enhancement of Clot Lysis In Vitro and In Vivo with a Bispecific Monoclonal Antibody Directed against Human Fibrin and against Urokinase-Type Plasminogen Activator
复制标题
使用针对人纤维蛋白和尿激酶型纤溶酶原激活剂的双特异性单克隆抗体增强体外和体内凝块溶解
DOI：
10.1055/s-0038-1646486
复制
发表时间：
1991
影响因子：
6.7
通讯作者：
D. Collen
中科院分区：
文献类型：
--
作者：
T. Kurokawa;S. Iwasa;A. Kakinuma;J. Stassen;H. Lijnen;D. Collen
将分泌人纤维蛋白特异性单克隆抗体（mAb）的小鼠杂交瘤与分泌抗尿激酶型纤溶酶原激活物（u-PA）单克隆抗体的小鼠杂交瘤融合，获得了一株分泌双特异性单克隆抗体（bs mAb）的杂交杂交瘤（FU1-74）。从小鼠腹水中纯化的单克隆抗体（MA-FU1-74）在未还原的SDS-PAGE上以单条带迁移，表观M r为150,000，对人纤维蛋白（K = 2 × 107 M - 1）和u-PA （K = 108 M - 1）具有亲和力，与从亲本杂交瘤中获得的单克隆抗体相当。ma - fu -74不影响双链u-PA （tcu-PA）对纤溶酶原或显色底物的酶活性。ma - fu -74与重组单链u-PA （rcu - pa）或与tcu-PA（尿激酶）的复合物分别使纤溶酶原激活剂对凝血人血浆的纤溶效力提高了20倍和5倍。在仓鼠肺栓塞模型中，与rscu-PA相比，rscu-PA/ ma - fu -74复合物的溶栓效力（每mg/kg u-PA溶栓百分比）提高了13至17倍。特异溶栓活性（u-PA抗原每εg/ml稳态血浆水平溶栓百分比）与rscu-PA无显著差异。rscu-PA与亲本抗u- pa单抗（MA-UK1-3）复合物的溶栓效力仅比rscu-PA增强2倍，特异性溶栓活性降低5倍。rscu-PA与ma - fu -74和MA-UK1-3复合物的血浆清除率均比rscu-PA低约10倍。在兔颈静脉血栓形成模型中，rscu-PA/MA-FU1-74复合物的溶栓效力增强4倍，特异性溶栓活性不变，血浆清除率降低20倍。在这两种动物模型中，rscu-PA/ ma - fu -74复合物对纤溶系统的激活并不比rscu-PA更广泛。由此可见，双特异性抗纤维蛋白/抗u-PA单克隆抗体MA-FU1-74可将u-PA靶向到纤维蛋白凝块上，从而显著增强纤溶酶原激活剂的溶栓效力。
Summary A hybrid hybridoma (FU1-74), secreting a bispecific monoclonal antibody (bs mAb), was obtained by fusion of a murine hybridoma secreting a monoclonal antibody (mAb) specific for human fibrin with a murine hybridoma secreting a mAb against urokinase-type plasminogen activator (u-PA). The bs mAb (MA-FU1-74), purified to homogeneity from mouse ascitic fluid, migrated as a single band with apparent M r 150,000 on non-reduced SDS-PAGE and had an affinity for both human fibrin (K a = 2 × 107 M–1) and for u-PA (K a = 108 M–1) comparable to that of the mAbs obtained from the respective parental hybridomas. MA-FU1-74 did not influence the enzymatic activity of two-chain u-PA (tcu-PA) towards plasminogen or towards a chromogenic substrate. The complex of MA-FU1-74 with recombinant single chain u-PA (rscu-PA) or with tcu-PA (urokinase) enhanced the fibrinolytic potency of the plasminogen activator towards clotted human plasma 20-fold and 5-fold, respectively. In a hamster pulmonary embolism model, the rscu-PA/MA-FU1-74 complex had a 13- to 17-fold increased thrombolytic potency (percent lysis per mg/kg u-PA administered) relative to that of rscu-PA. The specific thrombolytic activity (percent lysis per εg/ml steady state plasma level of u-PA antigen) of the complex was, however, not significantly different from that of rscu-PA. The complex of rscu-PA with the parental anti-u-PA mAb (MA-UK1-3) had only a 2-fold enhanced thrombolytic potency relative to that of rscu-PA and had a 5-fold decreased specific thrombolytic activity. The plasma clearance rates of the complexes of rscu-PA with both MA-FU1-74 and MA-UK1-3 were about 10-fold lower than that of rscu-PA. In a rabbit jugular vein thrombosis model, the rscu-PA/MA-FU1-74 complex had a 4-fold enhanced thrombolytic potency, an unchanged specific thrombolytic activity and 20-fold reduced plasma clearance. In both animal models, the rscu-PA/MA-FUl-74 complex did not cause more extensive systemic activation of the fibrinolytic system than rscu-PA. It is concluded that the bispecific anti-fibrin/anti-u-PA mAb MA-FU1-74 targets u-PA to the fibrin clot, resulting in a significantly enhanced thrombolytic potency of the plasminogen activator.
登录
查看更多内容
DOI：
10.1073/pnas.83.5.1453
发表时间：
1986-03-01
影响因子：
11.1
作者：
STAERZ, UD;BEVAN, MJ
通讯作者：
BEVAN, MJ
DOI：
--
发表时间：
1989
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Bode,C;Runge,MS;Branscomb,EE;Newell,JB;Matsueda,GR;Haber,E
通讯作者：
Haber,E
抗体-尿激酶缀合物的表征。
DOI：
--
发表时间：
1987
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Bode,C;Runge,MS;Newell,JB;Matsueda,GR;Haber,E
通讯作者：
Haber,E
影响因子：
56.9
作者：
HUI, KY;HABER, E;MATSUEDA, GR
通讯作者：
MATSUEDA, GR