Dickerson H W, Evans D L, Gratzek J B
Am J Vet Res. 1986 Nov;47(11):2400-4.
An initial panel of 34 hybridomas, each secreting antibodies reactive with an infective theront stage of an Ichthyophthirius multifiliis isolate, was produced. Three of these cell lines, each producing immunoglobulin M class antibodies, were cloned by limiting dilution and were expanded as ascites-producing tumors in syngeneic mice. Three monoclonal antibodies (MAB) reacted with intact whole theronts in an enzyme-linked immunosorbent assay to dilutions of 1:10,000 in ascitic fluids and had a similar pattern of surface and cytoplasmic staining in indirect immunofluorescent tests. Only antigen specified by MAB E6 could be characterized by acrylamide gel electrophoresis and immunoblotting; initial data indicated a molecular weight of 200,000. Physicochemical properties of the determinants recognized by the 3 MAB were tested by pronase digestion and periodate oxidation. Seemingly, a protein, glycoprotein, and carbohydrate were recognized by MAB E6, FE10, and AC8, respectively.
制备了一个初始的34种杂交瘤细胞株组成的细胞库，每个细胞株分泌的抗体都能与多子小瓜虫分离株的感染性幼虫期发生反应。其中三个细胞系，每个都产生免疫球蛋白M类抗体，通过有限稀释法进行克隆，并在同基因小鼠体内作为产生腹水的肿瘤进行扩增。在酶联免疫吸附试验中，三种单克隆抗体（MAB）与完整的幼虫在腹水稀释至1:10,000时发生反应，并且在间接免疫荧光试验中具有相似的表面和细胞质染色模式。只有MAB E6识别的抗原可以通过丙烯酰胺凝胶电泳和免疫印迹进行鉴定；初步数据表明其分子量为200,000。通过链霉蛋白酶消化和高碘酸盐氧化对三种MAB识别的决定簇的理化性质进行了测试。看起来，MAB E6、FE10和AC8分别识别一种蛋白质、一种糖蛋白和一种碳水化合物。
