Kinjoh K, Nakamura M, Kosugi T
Department of Physiology, School of Medicine, University of the Ryukyus, Okinawa, Japan.
Toxicon. 1994 Nov;32(11):1413-23. doi: 10.1016/0041-0101(94)90413-8.
We reported previously that habutobin, one of the type A thrombin-like enzymes, releases fibrinopeptide A alone from rabbit fibrinogen. To evaluate the effective action of habutobin in experiments using rabbit for the treatment of thrombosis, we attempted to develop an immunological method for measuring the fibrinopeptide A level in the circulating blood of rabbit. The purified rabbit fibrinopeptide A was coupled to keyhole limpet hemocyanin and BALB/c mice were immunized with the resultant fibrinopeptide A-hemocyanin conjugate. The spleen cells of an immunized mouse were fused with myeloma cells (P3-X63-Ag8-U1). As a result, one hybridoma (a-F-7) was selected, which secreted an antibody against rabbit fibrinopeptide A. Using this monoclonal antibody, we developed a competitive enzyme-linked immunoassay for estimating rabbit fibrinopeptide A. It was able to measure rabbit fibrinopeptide A contained in bentonite defibrinated plasma. This competitive enzyme-linked immunoassay should be useful for determining the fibrinopeptide A level in the circulating blood of rabbits, using plasma defibrinated by bentonite.
我们之前报道过，哈布托宾是一种A型类凝血酶，它能单独从兔纤维蛋白原中释放出纤维蛋白肽A。为了在兔实验中评估哈布托宾治疗血栓形成的有效作用，我们试图开发一种免疫方法来测定兔循环血液中的纤维蛋白肽A水平。将纯化的兔纤维蛋白肽A与钥孔戚血蓝蛋白偶联，并用所得的纤维蛋白肽A-血蓝蛋白偶联物免疫BALB/c小鼠。将免疫小鼠的脾细胞与骨髓瘤细胞（P3-X63-Ag8-U1）融合。结果，筛选出一株分泌抗兔纤维蛋白肽A抗体的杂交瘤（α-F-7）。利用该单克隆抗体，我们开发了一种竞争性酶联免疫分析法来测定兔纤维蛋白肽A。该方法能够测定膨润土去纤维蛋白血浆中所含的兔纤维蛋白肽A。这种竞争性酶联免疫分析法应用膨润土使血浆去纤维蛋白，对于测定兔循环血液中的纤维蛋白肽A水平应该是有用的。
