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用人高分子量尿激酶(HMW-UK)为抗原,通过杂交瘤技术获得了两株阳性杂交瘤细胞(2B8、3A2)。采用葡萄球菌 A 蛋白(SPA)亲和层析或 Water’s650快速蛋白分离系统纯化抗 UK 单克隆抗体(McAb).3A2McAb 为 IgG 1亚类,特异地识别 HMW-UK 和低分子量 LMW-UK,抑制 UK 活性,表明3A2McAb 所作用的位点为 UK 的 B 链,即活性中心。EIA 测定3A2McAb 与 UK 的亲和常数为8.43×10~8M~(-1)。2B_8McAb 亦为IgG_(?)亚类,特异地识别 HMW-UK 及其氨基端1～135氨基酸片段(ATF),不抑制活性,EIA 测定2B_8McAb 与UK 的亲和常数为2.8×10~9M~(-1)。2B_8McAb 可用于导向溶栓的研究以及 UK 与其受体间反应的研究。
Two positive hybridoma cells (2B8, 3A2) were obtained by hybridoma technique using human high molecular weight urokinase (HMW-UK) as antigen. Anti-UK monoclonal antibodies (McAbs) were purified using Staphylococcus A Protein (SPA) affinity chromatography or the Waters’s650 Rapid Protein Separation System.3A2McAb is an IgG subclass that specifically recognizes HMW-UK and low molecular weight LMW-UK, Inhibition of UK activity, indicating that the role of 3A2McAb site for the UK’s B chain, the active center. The affinity constant of 3A2McAb with UK measured by EIA was 8.43 × 10 ~ 8M ~ (-1). 2B_8McAb is also a subclass of IgG_ (?), Specifically recognizing HMW-UK and its amino terminal 1 to 135 amino acid fragments (ATF), does not inhibit the activity of EIA determination 2B_8McAb and UK affinity constant of 2.8 × 10 ~ 9M ~ -1). 2B_8McAb can be used to guide the study of thrombolysis and UK and its receptor reaction.