中国组织工程研究与临床康复2011，Vol.15Issue(24)：4557-4560,4.DOI:10.3969/j.issn.1673-8225.2011.24.046
纯化和标记抗人轻链β2m单克隆抗体的方法
Efficient purification and label of anti-human beta 2-microglobulin light chain monoclonal antibody
阮光萍 1姚翔 1庞荣清 1汪兴明 1戴莹 1潘兴华1
作者信息
- 1. 解放军昆明总医院,干细胞和组织器官工程研究中心,云南省昆明市,650032
- 折叠
摘要
Abstract
BACKGROUND: The inoculation of hybridoma cell strain onto mouse abdominal cavity may obtain ascites containing mass antibody. Previous method to purify monoclonal antibody in ascites is complex and difficult to operate.OBJECTIVE: To prepare, purify and label anti-human leukocyte antigen (HLA)-I class molecule light chain monoclonal antibody, and to detect the expression of tumor cell surface HLA-I class molecules. METHODS: Hybridoma cells were inoculated onto the mouse abdominal cavity. Ascites containing anti-human light chain beta2-microglobulin antibody were obtained and purified with the modified caprylic acid-ammonium sulfate method. The purified monoclonal antibody was labeled with fluorescein isothiocyanate to detect peripheral blood mononuclear cells, T2 cells expressing blank HLA-A2 molecule and K562 cells surface HLA-I class molecules. The expression of HLA-I class molecules was determined by using flow cytometry and fluorescent microscopy. RESULTS AND CONCLUSION: The purified anti-human light chain beta2-microglobulin-fluorescein isothiocyanate monoclonal antibodies accounted for 96% purity. Flow cytometry results showed that, the HLA-I class molecules were highly expressed in peripheral blood mononuclear cells surface, lowly expressed in T2 cells, and not expressed in K562 cells surface. It is a simple and convenient method to purify ascites with the modified caprylic acid-ammonium sulfate method, and according prepare anti-human light chain beta2-microglobulin-fluorescein isothiocyanate. This method is effective to distinguish the levels of HLA-I class molecules expressed in various cells.关键词
轻链/单克隆抗体/人类白细胞抗原Ⅰ/纯化/标记/组织工程分类
医药卫生引用本文复制引用
阮光萍,姚翔,庞荣清,汪兴明,戴莹,潘兴华..纯化和标记抗人轻链β2m单克隆抗体的方法[J].中国组织工程研究与临床康复,2011,15(24):4557-4560,4.
