人酸性成纤维细胞生长因子单克隆抗体的制备

Preparation of Monoclonal Antibodies Against Human Acidic Fi-broblast Growth Factor

以纯化的重组人酸性成纤维细胞生长因子(haFGF)为抗原,免疫BALB/c小鼠,取脾细胞与SP2/0细胞融合,获得一株稳定分泌抗haFGF的单克隆抗体的杂交瘤细胞株,其DNA含量为脾细胞与SP2/0细胞DNA含量之和,所分泌抗体为小鼠IgG_1亚类。免疫印迹显示:此单抗只与大肠杆菌中的haFGF有结合,与牛aFGF则没有结合反应。

BALB/c mice were immunized using purified recombinant human acidic fibroblast growth factor (haFGF) as antigen. Spleen cells from mice were fusde with SP2/0 cells and two hybridomas that could secret antibody against haFGF were cloned. The DNA contents of these two hybridomas was as the same as amount of spleen cells and SP2/0 cells, Isotype identification showed that both of these monoclonal antibodies were IgGi. Western-blot indicate that these antibodies only bound with haFGF among all proteins in E. coli, but not purified bovine acidic fibroblast growth factor.