重组人红细胞生成素单克隆抗体的制备和鉴定  

Preparation and Characterization of Monoclonal Antibodies to Recombinant Human Erythropoietin (rHuEPO)

作　　者：刘劲松[1] 史江[2] 刘慧[1] 房德兴[2] 伊纯德[1] 吴梧桐[3] 

机构地区：[1]解放军大连医学高等专科学校,大连116017 [2]解放军南京军区军事医学研究所,南京210002 [3]中国药科大学,南京210009

出　　处：《药物生物技术》1998年第3期137-139,共3页Pharmaceutical Biotechnology

摘　　要：以重组人红细胞生成素 (r Hu EPO)免疫 BALB/c小鼠 ,通过淋巴细胞杂交瘤技术将BALB/c小鼠脾细胞和 Sp2 /0骨髓瘤细胞融合 ,经 ELISA法筛选及有限稀释法克隆化 ,制备了 2株能稳定分泌抗 r Hu EPO单克隆抗体 (Mc Ab)的小鼠杂交瘤细胞系 G50和 A94。用鼠阊我隆抗体分型试剂盒鉴定 ,其分泌的 Mc Ab亚类均是 Ig G1 。经 ELISA鉴定 ,两株 Mc Ab只与 r Hu EPO特异性结合 ,与 IL- 1、GM- CSF、IFN- α和 IFN- r等细胞因子均无效叉反应。 Western- blot免疫印迹分析表明 ,两株 Mc Ab对 r HuBALB/c mice were immunized with the recombinant human erythropoietin (rHuEPO), The hybridoma cell lines were prepared by the conventional method. rHuEPO specific mouse monoclonal antibodies (McAbs) have been produced by two hybridoma cell lines named G50 and A94 respectively. The subclass of the McAbs were identified as IgG1 by using ELISA (mouse monoclonal antibody isotyping kit). Western blot results showed that the two McAbs recognized rHuEPO. ELISA test demonstrated that two McAbs did not have cross reaction with other cytokines, shus as Interleukin 2 (IL 2), Granunocyte maccrophage colonying stimulating factor (GM CSF), Interferon gamma (IFN r) and interferon alpha (IFN α).

关 键 词：重组 红细胞生成素 单克隆抗体 制备 鉴定 

分 类 号：R392－33[医药卫生—免疫学] R973.3[医药卫生—基础医学]