中华皮肤科杂志 ›› 2005, Vol. 38 ›› Issue (4): 208-210.

• 论著 • 上一篇    下一篇

天然属性的抗角蛋白单克隆IgM抗体的制备

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   

  1. 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
  • 收稿日期:2004-03-17 出版日期:2005-04-15 发布日期:2005-04-15
  • 通讯作者: 刘玉峰,E-mail:liuyuf@fmmu.edu.cn E-mail:liuyuf@fmmu.edu.cn
  • 基金资助:
    国家自然科学基金重点项目(30330510);军队重点基金项目(01Z086)

Preparation and Characterization of Hybridoma Strains Secreting Natural IgM Against Keratin

FU Meng1, LI Wei1, WANG Gang1, ZHANG Yan-guo2, LI Cheng-xin1, FAN Ping-shen1, LIU Yu-feng1   

  1. Department of Dermatology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China
  • Received:2004-03-17 Online:2005-04-15 Published:2005-04-15

PDF

268

赞

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。

关键词: 抗体,单克隆, 免疫球蛋白M, 杂交瘤, 角蛋白

Abstract: 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.

Key words: Antibodies, monoclonal, Immunoglobulin M, Hybridomas, Keratin