摘要
运用B淋巴细胞杂交瘤技术 ,以人内皮细胞特异性分子 1(ESM 1)蛋白为抗原 ,经PEG融合 ,有限稀释法筛选 ,获得了 7株稳定分泌抗人ESM 1的杂交瘤细胞株 ,其中 3A7细胞株特异性尤为显著 ,效价高达 1∶6 0 0 0。所分泌抗体亚型为IgG2b ,Westernblot结果表明对内皮细胞中的ESM 1蛋白及细胞培养上清中的ESM 1均可特异性识别。并观察到ESM 1主要分布在内皮细胞的细胞胞质伴胞核中。在肾组织中 ,ESM 1定位于血管内皮细胞 ,且肾癌标本中ESM 1阳性表达显著高于正常肾组织。免疫组化结果表明所获单抗具有一定的特异性和实用性。
The human endothelial cell specific molecule-1 (ESM-1) was used as antigen to prepare monoclonal antibodies by means of the B lymphocyte hybridoma technique. Seven strains of hybridoma cells secreting anti human ESM-1 stably were obtained after PEG fusion and screening by limited dilution,in which the McAb3A7 secreted by one of the hybridoma cells showed the highest specificity with the agglutination titer of 1∶6 000. The subtype of the secreted antibody was IgG2b. This monoclonal antibody could recognize specifically the ESM-1 both in endothelial cells and supernatants of cell cultures. The distribution of ESM-1 was mainly in the cytoplasm of endothelial cells,and located in the vasculoendothelial cells of renal tissues. Higher expression of ESM-1 in renal cell carcinoma than that of normal renal tissues was also noticed. These results indicate that this monoclonal antibody shows both high sensitivity and specificity.
出处
《现代免疫学》
CAS
CSCD
北大核心
2004年第3期217-220,共4页
Current Immunology
基金
国家自然科学基金资助项目 ( 3 9870 3 2 4)
安徽省人事厅跨世纪学术带头人研究基金
安徽省教育厅自然科学基金资助项目( 2 0 0 3kj184)
关键词
内皮细胞特异性分子-1
单克隆抗体
特性分析
免疫组化
肾癌
endothelial cell specific molecule-1
monoclonal antibody
characterization
immunohistochemistry
kidney neoplasms