Au D M, Kang A S, Murphy D J
Department of Biological Sciences, University of Durham, U.K.
Biochim Biophys Acta. 1989 Sep 11;1005(1):97-102. doi: 10.1016/0005-2760(89)90037-4.
Polyclonal antibodies raised against a range of seed apolipoproteins from the family Cruciferae have been used for the first time for low resolution epitope characterisation. Antibodies were raised against the major seed apolipoproteins of Brassica napus, Sinapis alba and Raphanus sativum. In each case, the antibodies recognized, in addition to the 19-20 kDa apolipoprotein to which they were raised, similar 19-20 kDa apolipoproteins from a wide range of species in the family Cruciferae, but not in other plant families. Homologous or heterologous two-sites (sandwich) assays were performed with the format [antibody A - test apolipoprotein - antibody B - 2 degrees antibody]. The results showed a drastically reduced antibody B binding by apolipoproteins preincubated with an antibody A. This indicated the presence of a single major epitope on many of the apolipoproteins. The antigenicity of native and denatured apolipoproteins was similar, although the antigenicity of the former was much more readily destroyed by proteinase attack. It is concluded that there are relatively few major epitopes present on the Cruciferae apolipoproteins and it is suggested that these epitopes are localized on the small hydrophilic surface-exposed C- and N-terminal domains of the apolipoproteins.
针对十字花科一系列种子载脂蛋白产生的多克隆抗体首次用于低分辨率表位表征。制备了针对甘蓝型油菜、白芥和萝卜主要种子载脂蛋白的抗体。在每种情况下，这些抗体除了能识别它们所针对的19 - 20 kDa载脂蛋白外，还能识别十字花科广泛物种中类似的19 - 20 kDa载脂蛋白，但不能识别其他植物科的载脂蛋白。采用[抗体A - 测试载脂蛋白 - 抗体B - 二抗]形式进行同源或异源双位点（夹心）测定。结果显示，与抗体A预孵育的载脂蛋白使抗体B的结合大幅减少。这表明许多载脂蛋白上存在单一主要表位。天然和变性载脂蛋白的抗原性相似，尽管前者的抗原性更容易被蛋白酶攻击破坏。结论是十字花科载脂蛋白上存在相对较少的主要表位，并且表明这些表位位于载脂蛋白小的亲水性表面暴露的C端和N端结构域。
