抗人内皮细胞特异性分子-1单克隆抗体的研制和初步鉴定

Preparation and Preliminary Identification of Monoclonal Antibodies Against Human Endothelial Cell Specific Molecule-1

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.