Functional characterization of monoclonal antibody inhibitors of alpha 2-antiplasmin that accelerate fibrinolysis in different animal plasmas.
Functional characterization of monoclonal antibody inhibitors of alpha 2-antiplasmin that accelerate fibrinolysis in different animal plasmas.
复制标题
α2-抗纤溶酶单克隆抗体抑制剂加速不同动物血浆中纤维蛋白溶解的功能表征。
DOI：
10.1089/hyb.1997.16.281
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发表时间：
1997
期刊：
影响因子：
--
通讯作者：
Reed,GL
中科院分区：
文献类型：
--
作者：
Reed,GL
在患有急性血栓性疾病的人类中，血栓常常表现出抵抗纤溶酶原激活剂诱导的纤溶作用。为了检查 α2-抗纤溶酶 (α2AP) 在体内血栓抵抗中的潜在作用，我们生成了 α2A P 的单克隆抗体抑制剂。在体细胞融合中，获得了 99 个杂交瘤，这些杂交瘤在捕获测定中产生与人 125I-α2AP 结合的 MAb。筛选分析显示，其中 3 个 MAb（49、70 和 77）中和了 α2AP 的功能。免疫印迹实验表明，这些 MAb 识别天然 α2AP 中存在的表位，该表位通过 SDS 变性而被破坏。这些 MAb 中的每一种均完全抑制其他 MAb 与 α2AP 的结合，但它们均不与另一种抗 α2AP MAb RWR 的结合竞争。当测试它们与血浆中的非人 α2AP 的结合时，所有三种 MAb 都与所有测试的灵长类动物血浆发生强烈交叉反应，但在其他血浆中显示出与 α2AP 结合的特殊模式，表明独特的精细表位特异性。在人血浆中，所有三种单克隆抗体均增强了纤溶酶原激活剂诱导的人血浆凝块的溶解，将尿激酶的效力提高了近 50 至 100 倍。这些单克隆抗体还显着增强了狒狒、食蟹猴、非洲绿猴血浆以及雪貂和狗血浆中凝块的溶解作用。由于它们能够有效抑制其他动物血浆中的 α2AP，这些 MAb 应可用于检查 α2AP 在体内血栓抗纤溶作用中的作用。
In humans with acute thrombotic disease, thrombi often appear to resist fibrinolysis induced by plasminogen activators. To examine the potential role of α2-antiplasmin (α2AP) in thrombus resistancein vivo, we generated monoclonal antibody inhibitors of α2A P. In a somatic cell fusion, 99 hybridomas were obtained that produced MAbs that bound to human125I-α2AP in a capture assay. Screening assays showed that 3 of these MAbs, 49, 70, and 77, neutralized the function of α2AP. Immunoblotting experiments indicated that these MAbs recognized an epitope present in native α2AP that was destroyed by denaturation with SDS. Each of these MAbs fully inhibited the binding of the other MAbs to α2AP, but none of them competed with the binding of another anti-α2AP MAb, RWR. When tested for their binding to nonhuman α2APs in plasmas, all three MAbs were strongly crossreactive with all primate plasmas tested but showed an idiosyncratic pattern of binding to α2AP in other plasmas, suggesting unique fine epitope specificities. In human plasma, all three MAbs amplified the lysis of human plasma clots induced by plasminogen activators, increasing the potency of urokinase by nearly 50- to 100-fold. These MAbs also markedly amplified the lysis of clots from baboon, cynomolgus, african green monkey plasmas, and to a lesser extent, ferret and dog. By virtue of their ability to potently inhibit α2AP in other animal plasmas, these MAbs should be useful for examining the role of α2AP in thrombus resistance to fibrinolysisin vivo.