Asakura S, Yoshida N, Matsuda M, Murayama H, Soe G
Institute of Hematology, Jichi Medical School, Tochigi, Japan.
Biochim Biophys Acta. 1988 Jan 4;952(1):37-47. doi: 10.1016/0167-4838(88)90099-4.
Monoclonal antibodies were raised against human thrombin-antithrombin III complex by a hybridoma technique. Among them, five monoclonal antibodies, designated as JITAT-4, -14, -16, -17 and -19, were found to react with thrombin-antithrombin III, but not with its nascent components, alpha-thrombin or antithrombin III. Their respective immunoglobulin classes are IgG1 for JITAT-16 and -19, and IgG2a for JITAT-4, -14 and -17. Besides the thrombin-antithrombin III complex, they all bound to the Factor Xa-antithrombin III complex and the active-site-cleaved two-chain antithrombin III as well. Moreover, the reactivity of these two antibodies to the neoantigens was not affected by heparin, suggesting that their epitopes are independent of heparin-induced conformational changes of antithrombin III. Two of them, JITAT-16 and -17, were categorized as high-affinity antibodies to thrombin-antithrombin III complex, the dissociation constants being 6.7 nM and 4.8 nM, respectively. However, they do not share antigenic determinants. These monoclonal antibodies may allow us to explore more precisely the reaction between antithrombin III and thrombin or its related enzymes.
通过杂交瘤技术制备了针对人凝血酶 - 抗凝血酶III复合物的单克隆抗体。其中，发现五种单克隆抗体，命名为JITAT - 4、-14、-16、-17和-19，可与凝血酶 - 抗凝血酶III反应，但不与其新生成分α-凝血酶或抗凝血酶III反应。它们各自的免疫球蛋白类别，JITAT - 16和-19为IgG1，JITAT - 4、-14和-17为IgG2a。除凝血酶 - 抗凝血酶III复合物外，它们还与因子Xa - 抗凝血酶III复合物以及活性位点裂解的双链抗凝血酶III结合。此外，这两种抗体对新抗原的反应性不受肝素影响，表明它们的表位独立于肝素诱导的抗凝血酶III构象变化。其中两种，JITAT - 16和-17，被归类为对凝血酶 - 抗凝血酶III复合物的高亲和力抗体，解离常数分别为6.7 nM和4.8 nM。然而，它们不共享抗原决定簇。这些单克隆抗体可能使我们能够更精确地探索抗凝血酶III与凝血酶或其相关酶之间的反应。
