天然属性的抗角蛋白单克隆IgM抗体的制备  

Preparation and Characterization of Hybridoma Strains Secreting Natural IgM Against Keratin

作　　者：付萌[1] 李巍[1] 王刚[1] 张衍国[2] 李承新[1] 樊平申[1] 刘玉峰[1] 

机构地区：[1]第四军医大学西京医院全军皮肤性病中心,西安710032 [2]第四军医大学唐都医院皮肤科

出　　处：《中华皮肤科杂志》2005年第4期208-210,共3页Chinese Journal of Dermatology

基　　金：国家自然科学基金重点项目(30330510);军队重点基金项目(01Z086)

摘　　要：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 natural anti-keratin IgM monoclonal antibody. Methods Spleen cells of BALB/c mice raised in specific pathogen free conditions were directly fused with Sp2/0 myeloma cells. The hybridoma supernatants were tested by ELISA using pre-extracted keratin. The natural IgM obtained was further identified by immunochemistry and immunoblot methods. Results The cell fusion rate was about 60% without pre-immunization. About 14% supernatants reacted with the keratin antigen. Three hybridoma strains secreting natural IgM monoclonal antibody against keratin were obtained. The immunochemistry results showed that the natural anti-keratin IgM was able to bind to epidermis, sebaceous gland, hair follicule, and muscle tissues. Conclusion B lymphocytes in normal BALB/c mice spleen can produce natural antibody against kerain.

关 键 词：IGM抗体 制备 抗角蛋白单克隆抗体 BALB/c小鼠 天然抗角蛋白 瘤细胞生长 无特殊病原体 免疫印迹技术 杂交瘤细胞株 免疫组化 蛋白抗原 ELISA SP2/0 B淋巴细胞 细胞融合 IGM型 自发分泌 小鼠脾脏 肌肉组织 脾细胞 上清液 

分 类 号：R392[医药卫生—免疫学]