内淋巴囊上皮细胞单克隆抗体的制作和特性

A study on monoclonal antibodies against epithelial cell of endolymphatic sac in guinea pigs

以显微解剖法取出的豚鼠内淋巴囊经超声波处理后，以其提取液作为原始抗原，探讨建立能够分泌识别内淋巴囊表面上皮细胞的单克隆抗体（ＭｃＡｂ）的系列杂交瘤。应用电镜免疫化学和Ｗｅｓｔｅｒｎ印迹法和聚丙酰胺凝胶电泳（ＳＤＳ－ＰＡＧＥ）测定，该抗体显示具有和内淋巴囊上皮细胞进行显著的免疫组织化学反应的活性，类似的反应出现在肾脏的近曲小管和亨氏袢的上皮细胞上，与其它１５个脏器（包括大脑、膀胱、胆囊、肺和肝脏等）无明显的交叉反应。所建立的单克隆抗体系列中，抗体亚型包括ＩｇＧ１、ＩｇＧ２ｂ、ＩｇＭ。此系列抗体分子量约为７４０００。该分子在内淋巴囊上皮细胞的构成和功能上的作用有待研究。

Using a microdissected endolymphatic sac of the guinea pig (n=35) as an initial antigen preparation, serials of monoclonal antibodies were established which were used to label the epithelial cells of endolymphatic sac. The antibodies showed strong immunoreactivity with kidney, but not with other organs . It also showed isotopes IgG1,IgG2b and IgM. The results of Western blotting and SDSPAGE indicated that the epitopes of monoclonal antibodies were proteins or glyoprotein with a molecular weight of approximately 7 400. The locations of the epitopes in epithelial cells suggest that it may play some roles in construction and function of the endolymphatic sac.