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目的制备天然属性的抗角蛋白单克隆IgM抗体。方法取饲养在无特殊病原体条件下BALB/c小鼠的脾细胞,直接与SP2/0细胞融合。以提取的角蛋白抗原对阳性杂交瘤细胞生长孔进行ELISA筛选。采用免疫组化、免疫印迹技术对获得的抗体进行鉴定。结果不经任何免疫,细胞融合率为60%;14%的杂交瘤细胞生长孔上清液与角蛋白抗原结合;获得3株稳定分泌IgM型天然抗角蛋白单克隆抗体的杂交瘤细胞株。免疫组化结果初步显示天然抗角蛋白IgM可以和表皮、皮脂腺、毛囊、肌肉组织结合。结论正常小鼠脾脏内存在自发分泌天然抗角蛋白IgM的B淋巴细胞。
Objective To prepare the natural properties of anti-keratin monoclonal IgM antibody. Methods Spleen cells of BALB / c mice fed without special pathogens were directly fused with SP2 / 0 cells. The positive hybridoma cell growth wells were subjected to ELISA screening with the extracted keratin antigen. The obtained antibodies were identified by immunohistochemistry and Western blotting. Results The cell fusion rate was 60% without any immunization. 14% of hybridoma cell growth supernatants were bound with keratin antigen. Three hybridoma cell lines stably secreting IgM-type natural keratin monoclonal antibodies were obtained. Immunohistochemical results preliminary showed that natural anti-keratin IgM and the epidermis, sebaceous glands, hair follicles, muscle tissue. Conclusion B lymphocytes secreting natural keratin IgM spontaneously exist in the spleen of normal mice.