Debus E, Weber K, Osborn M
EMBO J. 1983;2(12):2305-12. doi: 10.1002/j.1460-2075.1983.tb01738.x.
A set of monoclonal antibodies to desmin has been isolated from a fusion of mouse myeloma cells with spleen cells from mice immunized with purified porcine desmin. Eleven group I antibodies recognized desmin in the immune blot, and using defined desmin fragments the epitope has been tentatively assigned as lying between residues 325 and 372. When cell lines were tested in immunofluorescence only the human line RD and hamster BHK-21 were positive. When tissue sections were used, skeletal, cardiac, visceral and some vascular smooth muscle cells were positive. Thus, the group I antibodies appear specific for desmin and do not recognize other intermediate filament proteins. Group II monoclonals recognized not only desmin in the immune blot but also other polypeptides. The epitope of this class is located between residues 70 and 280. In immunofluorescence on cell lines and tissues, the staining patterns of group II antibodies were more complicated and demonstrate that not only other intermediate filament proteins but also additional antigenic determinants are being recognized. The group I antibodies stain, as expected from their desmin specificity, rat and human rhabdomyosarcomas and thus appear to be useful reagents in pathology.
一组抗结蛋白的单克隆抗体已从用纯化的猪结蛋白免疫的小鼠脾脏细胞与小鼠骨髓瘤细胞的融合物中分离出来。11种I组抗体在免疫印迹中识别结蛋白，通过使用特定的结蛋白片段，其表位已初步确定位于325至372位氨基酸残基之间。当在免疫荧光中检测细胞系时，只有人源细胞系RD和仓鼠BHK - 21呈阳性。当使用组织切片时，骨骼肌、心肌、内脏和一些血管平滑肌细胞呈阳性。因此，I组抗体似乎对结蛋白具有特异性，不识别其他中间丝蛋白。II组单克隆抗体不仅在免疫印迹中识别结蛋白，还识别其他多肽。该类别的表位位于70至280位氨基酸残基之间。在细胞系和组织的免疫荧光中，II组抗体的染色模式更为复杂，表明不仅识别其他中间丝蛋白，还识别额外的抗原决定簇。正如预期的那样，I组抗体因其对结蛋白的特异性而能使大鼠和人横纹肌肉瘤染色，因此似乎是病理学中有用的试剂。
