摘要
用重组人促红细胞生成素 (rhEPO)免疫Balb c小鼠 ,取其脾细胞在PEG4 0 0 0作用下与SP2 0小鼠骨髓瘤细胞融合 ,获得一株能分泌抗rhEPO单抗隆抗体的杂交瘤细胞株 2F12 ,染色体数目大于 10 0条 ,间接ELISA法测定腹水和细胞培养上清效价 ,分别为 1.6× 10 -7和 4× 10 -4 。测定抗体亚类时 ,则同时显示IgA和IgG1,其轻链为κ链 ;相对亲和力为 5× 10 -12 mol L。单抗 2F12腹水经纯化获高纯度抗体 ,并用此抗体制备了抗体亲和色谱柱 。
rhEPO was used to immunize Bal b/c mice and a hybridoma cell line 2F12 was established by fusing SP2/0 myeloma cells with the immunized mouse spleen cells. The hybridom cell chromosome were more than 100. 2F12 was shown to be IgG1 and IgA and light chain was κ by indirect ELISA test. 2F12 McAb showed strong reaction with rhEPO, but no reaction with CHO cell proteins. The titer of ascitic fluid and cells supernatant fluid was 1.6×10 -7 and 4×10 -4 respectively. The affinty was 5×10 -12 mol/L. An antibody affinity chromatography column was preparizd with 2F12 IgG and purified rhEPO.
出处
《生物技术通讯》
CAS
2000年第3期209-212,共4页
Letters in Biotechnology
