一株抗重组人促红细胞生成素单克隆抗体杂交瘤的研制

Preparation of A Monoclonal Antibody Against Reconbinant Human Erythropoietin

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

Objective To prepare monoclonal antibody (McAb) against reconbinant human erythropoietin (rhEPO) and identify the biological features of the McAb.Methods BALB/c mice were immunized with rhEPO by routine methods.The hybridomas were obtained by cell fusion.The ELISA assay and limited diluting subcloning methods were used to select the positive clones.The McAb was identified by Western blotting and quickly Immunoglobin subclass assay.Results The hybridoma (1D1) obtained was able to permanently secret anti-rhEPO McAb of which the isotype was IgG2b with κ light chains.The McAb could specifically identify the rhEPO that was able to be verified by Western blotting assay.Conclusion A hybridoma which could permanently secret anti-rhEPO McAb was successfully obtained.