目的建立重组人红细胞生成素二聚体工程细胞株。方法用重组人红细胞生成素二聚体表达载体转染COS-7细胞,通过dot blot进行检定。然后对CHO-dhfr-细胞进行稳定转染,再经过稀释克隆和MTX的加压扩增,筛选二聚体工程细胞株并进行传代及无血清培养基培养、目的蛋白纯化及检测。结果瞬时转染70h细胞有目的蛋白表达。稳定转染后选取表达量最高的细胞株为工程细胞株。该细胞株连续传15代及经无血清培养基培养,目的蛋白表达量均无明显下降。所表达的目的蛋白相对分子质量为70000,并具有高度特异性。结论已获得稳定高效表达重组人红细胞生成素二聚体的工程细胞株。 Objective To establish a recombinant human erythropoietin dimer engineering cell line. Methods COS-7 cells were transfected with recombinant human erythropoietin dimer expression vector and verified by dot blot. The CHO-dhfr- cells were then stably transfected, and then diluted and cloned by MTX. The dimer-engineered cell lines were screened and subcultured in serum-free medium for purification and detection. Results The target protein was transiently transfected for 70 hours. Stable transfected cells selected the highest expression of the engineering cell lines. The cell line continuously passed 15 generations and cultured in serum-free medium, the expression of the target protein were no significant decline. The expressed protein of interest has a relative molecular mass of 70,000 and is highly specific. Conclusion The stable and highly expressed recombinant human erythropoietin dimer has been obtained.