Yang C C, Chan H L
Department of Life Science, National Tsing Hua University, Hsinchu, Taiwan.
Toxicon. 1999 May;37(5):729-42. doi: 10.1016/s0041-0101(98)00213-x.
The number of antigenic determinants of beta1-bungarotoxin (beta1-Bgt), A chain and B chain were determined to be seven, five and two, respectively, by quantitative precipitin reactions and analysis of the soluble complex formed from beta1-Bgt and Fab fragments of the antibody. The gel filtration patterns on a size exclusion column revealed that the soluble complexes formed from non-precipitating antibody and beta1-Bgt at a different molar ratio all emerged in the void volume, indicating that the molecular weight of the soluble complex is around 6000 kDa or more. The gel filtration pattern from Fab fragments of non-precipitating antibody also revealed that only 1 or 2 molecules of Fab fragments are bound to beta1-Bgt, A chain and B chain instead of seven, five and two molecules as in the case of precipitating antibody. Besides, twenty-three stable monoclonal antibodies (mAbs) were prepared against beta1-Bgt by the hybridoma technique. Most of these mAbs cross-reacted with isotoxins of the beta-Bgt family, as determined by an enzyme-linked immunosorbent assay, but none of them reacted with notexin, notechis II-5 and three PLA2 homologues, PLA2, DE-I and CMS-9.
通过定量沉淀反应以及对由β1-银环蛇毒素（β1-Bgt）与抗体的Fab片段形成的可溶性复合物进行分析，确定β1-银环蛇毒素、A链和B链的抗原决定簇数量分别为7个、5个和2个。在尺寸排阻柱上的凝胶过滤图谱显示，由非沉淀抗体与β1-Bgt以不同摩尔比形成的可溶性复合物均出现在空体积中，这表明可溶性复合物的分子量约为6000 kDa或更大。非沉淀抗体的Fab片段的凝胶过滤图谱还显示，与沉淀抗体的情况不同，只有1个或2个Fab片段分子与β1-Bgt、A链和B链结合。此外，通过杂交瘤技术制备了23种针对β1-银环蛇毒素的稳定单克隆抗体（mAb）。通过酶联免疫吸附测定确定，这些mAb中的大多数与β-Bgt家族的同毒素发生交叉反应，但它们均未与海蛇毒素、虎蛇毒素II-5以及三种磷脂酶A2同源物（磷脂酶A2、DE-I和CMS-9）发生反应。
