Delmotte F, Brillouet C, Leclerc M, Luquet G, Kader J C
Eur J Immunol. 1986 Nov;16(11):1325-30. doi: 10.1002/eji.1830161103.
Cells from sea star (Asterias rubens) axial organs stimulated with trinitrophenyl (TNP) or fluoresceinyl-haptened polyacrylamide beads and subsequently stimulated in vitro with the same antigen produced and released a specific antibody-like protein which induced lysis of haptened sheep erythrocytes in the presence of serum complement. The anti-TNP antibody-like protein isolated by ammonium sulfate precipitation, gel filtration and affinity chromatography exhibited a single precipiting peak after crossed immunoelectrophoresis against rabbit antiserum to partially purified culture supernatant. The anti-TNP antibody-like protein gave a specific affinity precipitate in crossed affino-electrophoresis using a p-nitrobenzoyl-substituted gel. The analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under both reducing and nonreducing conditions evidenced a unique 30-kDa polypeptide chain. According to gel filtration experiments, the molecular weight of the major component isolated by affinity chromatography was about four times higher. Therefore, the antibody-like molecule could be a tetrameric protein devoid of any disulfide bond.
用三硝基苯基（TNP）或荧光素标记的聚丙烯酰胺珠刺激海星（Asterias rubens）轴器官的细胞，随后在体外用相同抗原刺激，产生并释放出一种特异性抗体样蛋白，该蛋白在血清补体存在下可诱导致敏绵羊红细胞裂解。通过硫酸铵沉淀、凝胶过滤和亲和层析分离得到的抗TNP抗体样蛋白，在与兔抗血清进行交叉免疫电泳以检测部分纯化的培养上清液时，呈现出单一沉淀峰。抗TNP抗体样蛋白在使用对硝基苯甲酰取代凝胶的交叉亲和电泳中产生特异性亲和沉淀。在还原和非还原条件下进行的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析表明，存在一条独特的30 kDa多肽链。根据凝胶过滤实验，通过亲和层析分离得到的主要成分的分子量约高四倍。因此，抗体样分子可能是一种不含任何二硫键的四聚体蛋白。
