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目的 研制特异性识别人促红细胞生成素的单克隆抗体 ,并对其生物学特性作出初步鉴定。方法 采用重组人促红细胞生成素 (rhEPO)为抗原 ,常规免疫BALB/c小鼠 ,经细胞融合、ELISA筛选、有限稀释亚克隆获得抗人EPO杂交瘤 ,以免疫印迹和快速Ig亚型分类对所获单抗作出初步鉴定。结果 经筛选获得 1株稳定分泌的抗rhEPO单抗的杂交瘤细胞株 (1D1)。亚型鉴定为IgG2b ,轻链为κ链 ,用Westernblot分析证实该单抗对rhEPO特异性识别。结论 成功获得 1株可分泌特异性识别rhEPO的单克隆抗体的杂交瘤
Objective To develop a monoclonal antibody that specifically recognizes human erythropoietin and to make preliminary identification of its biological characteristics. Methods Recombinant human erythropoietin (rhEPO) was used as antigen to immunize BALB / c mice. The anti-human EPO hybridomas were subcloned by means of cell fusion, ELISA and limited dilution. The immunoblot and rapid Ig subtype The monoclonal antibody was initially identified. Results A strain of monoclonal antibody against rhEPO stably secreting monoclonal antibody (1D1) was screened. The subtype was identified as IgG2b and the light chain was kappa chain. Western blot analysis confirmed the specificity of the mAb for rhEPO. Conclusion A hybridoma secreting monoclonal antibodies that specifically recognize rhEPO was successfully obtained