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目的：真核表达及纯化重组人凝血因子Ⅶ（rFⅦ）。方法通过PCR方法扩增FⅦ全长cDNA，克隆入真核表达载体pcDNA3.0，利用脂质体将FⅦ基因与二氢叶酸还原酶基因共转染CHO-dhfr-/-细胞，采用氨甲喋呤药物加压，Western blot方法筛选获得稳定高表达细胞克隆。将高表达细胞克隆在摇瓶中无血清悬浮驯化培养，收集细胞培养上清，通过离子交换和亲和层析两步法进行纯化并对纯化产物进行活性鉴定。结果在CHO细胞中成功表达了rFⅦ，无血清悬浮培养条件下表达量可达3 mg?L-1?d-1，成功纯化了rFⅦ，SDS-PAGE凝胶电泳显示分子质量约50 kDa的单一多肽，生物学活性试验证实该纯化产物具有促凝活性。结论成功表达并纯化了rFⅦ，为进一步研制活化凝血因子Ⅶ药物奠定了基础。“,”Objective Eukaryotic expression and purification of recombinant human coagulation factorⅦ. Methods Human FⅦcDNA was amplified by PCR and cloned into pcDNA3.0 expression vector. FⅦexpression plasmid and DHFR expression plasmid were co-transfected by lipofectamine to CHO-dhfr-/-cells. Stable high expression level cell clones were screened under MTX pressure by Western blot assay. After serum-free and suspension adaption culture, cell supernatant was collected and the recombinant FⅦwas purified by ion-exchange and affinity chromatography. The biological activity of the purified recombinant FⅦ was assayed. Results Recombinant human FⅦwas successfully expressed in CHO cells, the expression level was about 3 mg?L-1?d-1 under serum-free and suspension condition. Recombinant human FⅦwas successfully purified as a 50 kDa single chain peptide and identified bioactive. Conclusion Recombinant human coagulation factorⅦwas successfully expressed and purified. This work is important for further study of active FⅦdrug.