抗爪蟾角蛋白单克隆抗体的研究

本实验以非洲爪蟾(Xenopus laevis)成体皮肤角蛋白免疫的BALB/c鼠脾细胞与小鼠Sp2/0骨髓瘤细胞在PEG的作用下进行融合,经HAT选择培养、ELISA筛选、反复克隆化及冻存复苏,获得四株稳定分泌抗体的杂交瘤细胞株。对其中一株杂交瘤细胞KD_(10)及其分泌的抗体进行了较全面的研究。

Study on Monoclonal Antibody against Frog (Xenopus laevis) Epidermal Keratin

The hybridoma KD10 producing monoclonal antibody against frog (Xenopus laevis) epidermal keratins have been established by fusing mouse myeloma cell line Sp2/0 with psnlee cell from a BALB/c mouse immunized with keratin extract of the epidermis of adult frog. Which can secret McAb against keratin stably. Tissue sections of Xenopus and human tumor as well as apparently normal tissue were tested with immunoperoxidase reaction. The McAb KD10 reacted strongly with epithelial cell of all tissues, but not reacted with non-epithelial cells, McAb KD10 could react with human tumor cells orgina-ted from epithelia,for example,breast carcinoma and colon carcinoma. None reacted with tumor cells of non-epithelia origin. This McAb ascite fluids was purified by Protein A-Sepharose CL-4B. McAb KD10 is determed as IgG1 subclass with molecular weight of 180,000 delton. It is a "width spectum" McAb against keratin. The antigen (keratin) is a heat-stable protein. Its antigenicity is not damaged by treatment of acid and base of low concentration or by oxidant and ethenol.