Coppola R, Tombesi S, Valentini F, Alborali S, Albertini A, Mannucci P M
Angelo Bianchi Bonomi Hemophilia and Thrombosis Center, IRCCS Maggiore Hospital, Milano, Italy.
Thromb Res. 1992 Nov 1;68(3):269-81. doi: 10.1016/0049-3848(92)90084-n.
Eight murine monoclonal antibodies to human factor VII have been produced and characterized. As tested by direct ELISA, all antibodies bound to immobilized factor VII and this interaction did not require Ca(II). On immunoblotting three antibodies reacted only with native factor VII and activated factor VII (rFVIIa), two with native factor VII, activated factor VII and with the light chain of reduced activated factor VII and three with reduced activated factor VIIa (light chain) only. When coupled to Sepharose five antibodies were capable of immunodepleting factor VII from normal plasma and inhibited coagulant activity with varied potencies. Among these antibodies, one (10C12.2) had peculiar characteristics, lacking reactivity with Gladomainless FVII and with the Ca(II)-dependent conformation. This antibody has been used to develop a two-site ELISA which appears to measure native factor VII but not poorly carboxylated, inactive forms of the molecule.
已制备并鉴定了八种针对人凝血因子VII的鼠单克隆抗体。通过直接ELISA检测，所有抗体均与固定化的凝血因子VII结合，且这种相互作用不需要Ca(II)。在免疫印迹中，三种抗体仅与天然凝血因子VII和活化凝血因子VII（rFVIIa）反应，两种与天然凝血因子VII、活化凝血因子VII以及还原活化凝血因子VII的轻链反应，三种仅与还原活化凝血因子VIIa（轻链）反应。当与琼脂糖偶联时，五种抗体能够从正常血浆中免疫去除凝血因子VII，并以不同的效力抑制凝血活性。在这些抗体中，一种（10C12.2）具有独特的特性，与无Gla结构域的FVII和Ca(II)依赖性构象无反应性。该抗体已用于开发一种双位点ELISA，该方法似乎可检测天然凝血因子VII，但不能检测该分子羧化不良的无活性形式。
