Nakamura M, Hanashiro K, Kosugi T
Department of Physiology, School of Medicine, University of Ryukyus, Okinawa, Japan.
Toxicon. 1993 Oct;31(10):1325-8. doi: 10.1016/0041-0101(93)90405-8.
A monoclonal antibody to the thrombin-like enzyme, habutobin, was produced. An ELISA-double sandwich method employing this monoclonal antibody was devised as a method for determining the habutobin concentration in vitro. However, the absorbance of the control sample in such an ELISA-double sandwich procedure was too high to estimate low levels of the enzyme. The present study therefore attempted to establish a reliable ELISA-double sandwich method in which the absorbance of the control sample was lower than previously, and which had a high sensitivity, in order to determine the habutobin concentration in vitro and in vivo. The modification of the ELISA-double sandwich technique employing the monoclonal antibody against habutobin, the purified rat IgG against habutobin and POD-mouse anti-rat IgG2a, provided a reliable means of determining the habutobin levels in the circulating blood of rabbits.
制备了一种针对类凝血酶哈布毒素的单克隆抗体。设计了一种采用该单克隆抗体的酶联免疫吸附测定 - 双夹心方法，作为体外测定哈布毒素浓度的方法。然而，在这种酶联免疫吸附测定 - 双夹心程序中，对照样品的吸光度过高，无法估计低水平的该酶。因此，本研究试图建立一种可靠的酶联免疫吸附测定 - 双夹心方法，其中对照样品的吸光度比以前更低，且具有高灵敏度，以便在体外和体内测定哈布毒素浓度。采用针对哈布毒素的单克隆抗体、纯化的抗哈布毒素大鼠IgG和过氧化物酶标记的小鼠抗大鼠IgG2a对酶联免疫吸附测定 - 双夹心技术进行改进，提供了一种可靠的方法来测定兔循环血液中的哈布毒素水平。
