人源抗_HBsAgdsFv_IFN融合基因的构建及在CHO细胞中的表达.pdfVIP

2002 3 26 1 32 Bull Acad Mil Med Sci, Mar 2002; Vol 26 o 1 _HBsAg dsFv_IFN CHO , , , , , , ( 军事医学科学院微生物流行病研究所, 北京 100071) [ ] :探讨用抗体工程技术研制乙型肝炎导向干扰素:在已有的抗HBsAg_dsFv 的基础上, 用重叠延 伸PCR 的 法将抗_HBsAg_dsFv V 基因和A_干扰素基因连接,形成融合基因为避免IF 与dsFv 空间折叠的影响, L 其间引入 15 个氨基酸的柔性短肽; 分别构建重链V 基因轻链V _IF 融合基因的 CHO 表达载体, 共转染CHO 细 H L 胞:从细胞培养上清中检测到有抗病毒活性的IF 存在, 有抗_HBsAg 活性, 但dsFv 与抗原的结合活性较低 : 本研究为研制乙肝导向干扰素奠定了基础 [ ] 肝炎,乙型; 二硫键抗体;干扰素; 真核表达;CHO 细胞 [ ] Q78; R512.6 [ ] A [ ] 1000_5501(2002)01_0032_03 Construction of anti_HBsAg dsFv_IFN fusion gene and its eukaryotic expression in CHO cells YU Chang_Ming, CHEN Wei, X UJing, FULing, SONG Hong_Bin, TONG Yi_Gang, WAN GHai_Tao ( Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences,Beijing 100071,China) [ Abstract] Ob ective: To study the feasibility of expressing human anti_HBsAg_dsFv_IF in CHO cells as a method for preparing the putative targeting drug for hepatitis B.Methods:Based on the constructed anti_ HBsAg_dsFv gene, the V genewas L fusedwith IF _A2a gene by overlapping_PCR. V _IF and V were cloned to different mammalian expression vectors respectively. L H _ These vectors were cotransfected into CHO/ dhf r cells.The supernatant of cultured transfected cells was collected and assayed for IF activity and HBsAg affinity by ELISA. Results:The supernatant exhibited anti_HBsAg activity and IF activity. Conclusions: The successful expr