›› 2010, Vol. 30 ›› Issue (4): 289-292.

• 论文 • 上一篇    下一篇

激发型鼠抗人4-1BBL单克隆抗体的研制

  

  1. 苏州大学实验动物中心,苏州,215123
  • 收稿日期:2010-02-01 出版日期:2010-04-30 发布日期:2010-04-30

Preparation of Monoclonal Antibody Against Human 4-1BBL

  1. Laboratory Animal Center, Suzhou University,Suzhou 215123,China
  • Received:2010-02-01 Online:2010-04-30 Published:2010-04-30

摘要: 目的 鼠抗人4-1BBL单克隆抗体的制备及其生物学特性研究。方法 以高表达人4-1BBL分子的基因工程细胞株L929/4-1BBL为免疫原免疫BALB/c小鼠,利用淋巴细胞杂交瘤技术进行细胞融合,经过选择性克隆化培养及流式细胞术分析,筛选分泌特异性抗人4-1BBL单抗的杂交瘤细胞株;采用快速定性试纸法鉴定单抗亚类、间接免疫荧光法检测单抗对肿瘤细胞株的识别谱。进而,将所获抗人4-1BBL单抗体外刺激白血病细胞株U937,通过细胞计数法分析单抗对U937生长的影响。结果 获得一株持续、稳定分泌鼠抗人4-1BBL单抗的杂交瘤细胞株,并将其分泌的单抗命名为39A8.单抗39A8可有效识别不同肿瘤细胞株表达的4-1BBL,经细胞计数分析,39A8对U937细胞的生长具有明显的促增殖作用。结论 单抗39A8是可激发4-1BBL逆向信号的单克隆抗体,是研究4-1BBL分子及其功能的重要工具。

关键词: 4-1BBL, 杂交瘤, 单克隆抗体

Abstract: Objective To prepare monoclonal antibodies against human 4-1BBL molecule and analyze their biological characteristics. Methods The BALB/c mice were immunized with the cell line L929/4-1 BBL. The splenocytes from the immunized mice were fused with SP2/0, and then the hybridoma cells were screened with immunostaing and FACS. The isotype of the monoclonal antibody was identified by rapid isotyping analysis, and the 4-1 BBL expression profile on tumor cell lines was analyzed by indirect immunofluorescence with obtained mAb 39A8. Furthermore, the myeloid leukemia cell line U937 cells were stimulated with mAb 39A8 and their proliferation was measured with cell number counting. Results A hybridoma cell line named 39A8 was obtained, which had the property of secreting anti-human 4-1 BBL monoclonal antibody continuously and stably. The mAb 39A8 could recognize 4-1 BBL molecules on different cell lines and could enhance the proliferation of U937 cells in dose-dependent manner. Conclusion This work established an agonistic anti-human 4-1BBL mAb which may provide a useful tool for studying 4-1 BBL and its biological function.

Key words: 4-1 BBL, Hybriboma, Monoclonal antibody