Imhof B A, Vollmers H P, Goodman S L, Birchmeier W
Cell. 1983 Dec;35(3 Pt 2):667-75. doi: 10.1016/0092-8674(83)90099-5.
With functional assays we selected a monoclonal antibody (anti-Arc-1) that perturbs specific cell-cell contacts of MDCK epithelial cells in tissue culture. The cells loosen their tight junctions, become uncoupled, and their polarity is abolished. Several findings show that the corresponding antigen appears to be a cell-surface protein of MDCK epithelial cells but not of other established cell lines: radioactively labeled antibody could be absorbed on the surface of intact cells; immunoscanning electron microscopy showed a homogeneous surface distribution of the antigen; and the antigen was extractable from the cells by EDTA. Absorption on Western blots and gel filtration showed antigenicity at 130 and 40 kd. This study shows that the hybridoma technique, in combination with functional screening of the monoclonal antibodies, allows the identification of new and specific molecules involved in cell-cell adhesion.
通过功能分析,我们筛选出一种单克隆抗体(抗Arc-1),它能在组织培养中扰乱MDCK上皮细胞的特定细胞间接触。细胞的紧密连接松弛,细胞间解偶联,极性消失。多项研究结果表明,相应抗原似乎是MDCK上皮细胞而非其他已建立细胞系的细胞表面蛋白:放射性标记抗体可被完整细胞表面吸收;免疫扫描电子显微镜显示抗原在细胞表面均匀分布;并且该抗原可被EDTA从细胞中提取出来。Western印迹和凝胶过滤的吸收实验显示,抗原性出现在130和40kd处。本研究表明,杂交瘤技术与单克隆抗体的功能筛选相结合,能够鉴定出参与细胞间黏附的新的特异性分子。