摘要
目的建立能够分泌特异性抗小鼠endoglin的单克隆抗体杂交瘤细胞系,为深入研究endoglin的新功能、诊断和治疗肿瘤血管生成提供物质基础。方法利用小鼠endoglin胞外段重组蛋白作为抗原免疫大鼠,通过细胞融合和杂交瘤筛选技术建立特异分泌抗小鼠endoglin单克隆抗体的杂交瘤系。利用酶联免疫吸附方法确定单克隆抗体亚型,免疫印迹方法明确筛选的单克隆抗体的特异性,免疫组化和免疫荧光初步判定获得的单克隆抗体是否可以应用于临床诊断。结果通过对90多个分泌抗体的融合细胞克隆的筛选,获得一株分泌特异性抗小鼠endoglin的单克隆抗体杂交瘤细胞系,分泌的单克隆抗体亚型为IgG1,轻链亚型为kappa。免疫印迹显示该研究获得的单克隆抗体可以特异识别小鼠endoglin重组蛋白,该单克隆抗体可以结合实体肿瘤组织微血管上的特异抗原并将微血管有效染色。结论本研究所获得的抗小鼠endoglin单克隆抗体将有助于深入研究endoglin的新功能,为肿瘤血管生成的诊断和治疗提供了物质保障。
[Objective] To establish a strain of hybridoma cell line secreting monoclonal antibody against murine endoglin. [Methods] A recombinant murine endoglin protein was used as antigen, and standard cell fusion and screening technology was used to establish the hybridoma cell line. The subtype of the monoclonal antibody was detected by enzyme linked immunosorbent assay. The specificity of the monoclonal antibody against murine endoglin was tested by Western blot. The monoclonal antibody was further proved to work by immunohistochemistry and immunofluorescence. [Results] More than 90 antibody-producing clones were screened. One clone, designated as EDG-1, was proved to have the capability of stably secreting monoclonal antibody against murine endoglin. The subtype of EDG-1 monoclonal antibody was IgG1, and the light chain was kappa. EDG-1 monoclonal antibody well recognized the recombinant murine endoglin in Western blot. Microvesseles in the sections of CT26 tumor tissues incubated with EDG-1 monoclonal antibody showed clear staining in a standard morphological state by immunohistochemistry and immunofluorescence. [Conclusionl EDG-1 monoclonal antibody may be a useful tool for further studying the new functions of endoglin and for diagnosis and treatment of tumor angiogenesis in solid tumors of murine models.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2007年第1期1-4,共4页
China Journal of Modern Medicine
基金
This study was supported by National Natural Science Foundation of China (No.30360115 & No.30560048), by Program for New Century Excel-lent Talents in University of China and by Education Department of Hainan Province (Hjkj200422)
