Objective:To express the soluble recombinant hemangiopoietin protein in E.coli BL21(DE3).Methods:Using human fetal live cDNA as a template,a partial cDNA fragment of HAPO coding N-terminal region was subcloned into pl...Objective:To express the soluble recombinant hemangiopoietin protein in E.coli BL21(DE3).Methods:Using human fetal live cDNA as a template,a partial cDNA fragment of HAPO coding N-terminal region was subcloned into plasmids pTrc99,pQE60 and pET32c to construct different recombinant prokaryotic expression systems.After selecting,the soluble rhHAPO fusion protein was expressed stably in E.coli BL21(DE3) by vector pET32c-HAPO and further isolated by nickelnitrilotriacetic acid(NTA) affinity chromatography.After cleavage with enterokinase,the rhHAPO protein was applied to Fast Flow SP sepharose column.Results:The rhHAPO protein had a purity of more than 95% and a good bioactivity based on the cell adhesion assay in ECV304 cells.Conclusion:We have established a protein engineering system to produce rhHAPO which may provide the possibility for clinical application.展开更多
目的:探讨重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白(rhTNFR:Fc)治疗难治性类风湿关节炎的临床疗效。方法:50例难治性类风湿关节炎(refractory or resistant rheumatoid arthritis,RRA)随机分为治疗组25例和对照组25例。治疗组:益赛普25...目的:探讨重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白(rhTNFR:Fc)治疗难治性类风湿关节炎的临床疗效。方法:50例难治性类风湿关节炎(refractory or resistant rheumatoid arthritis,RRA)随机分为治疗组25例和对照组25例。治疗组:益赛普25mg,皮下注射,每周2次,疗程6个月;对照组:安慰剂25mg,皮下注射,每周2次,疗程24周。疗效评价采用美国风湿病学会(ACR)疗效评定标准。结果:治疗组ACR20、ACR50、ACR70有效率均高于对照组(P<0.05),治疗组24周ACR20、ACR50、ACR70的有效率高于治疗组4周(P<0.05);治疗组发生不良事件6例(24.0%)高于对照组3例(12.0%)。结沦:rhTNFR:Fc治疗难治性类风湿关节炎有较好的疗效。展开更多
基金the Natural Science Foundation of China (30300186)the Grant of 863 projects from the Ministry of Science & Technology of China (2002AA223354)
文摘Objective:To express the soluble recombinant hemangiopoietin protein in E.coli BL21(DE3).Methods:Using human fetal live cDNA as a template,a partial cDNA fragment of HAPO coding N-terminal region was subcloned into plasmids pTrc99,pQE60 and pET32c to construct different recombinant prokaryotic expression systems.After selecting,the soluble rhHAPO fusion protein was expressed stably in E.coli BL21(DE3) by vector pET32c-HAPO and further isolated by nickelnitrilotriacetic acid(NTA) affinity chromatography.After cleavage with enterokinase,the rhHAPO protein was applied to Fast Flow SP sepharose column.Results:The rhHAPO protein had a purity of more than 95% and a good bioactivity based on the cell adhesion assay in ECV304 cells.Conclusion:We have established a protein engineering system to produce rhHAPO which may provide the possibility for clinical application.
文摘目的:探讨重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白(rhTNFR:Fc)治疗难治性类风湿关节炎的临床疗效。方法:50例难治性类风湿关节炎(refractory or resistant rheumatoid arthritis,RRA)随机分为治疗组25例和对照组25例。治疗组:益赛普25mg,皮下注射,每周2次,疗程6个月;对照组:安慰剂25mg,皮下注射,每周2次,疗程24周。疗效评价采用美国风湿病学会(ACR)疗效评定标准。结果:治疗组ACR20、ACR50、ACR70有效率均高于对照组(P<0.05),治疗组24周ACR20、ACR50、ACR70的有效率高于治疗组4周(P<0.05);治疗组发生不良事件6例(24.0%)高于对照组3例(12.0%)。结沦:rhTNFR:Fc治疗难治性类风湿关节炎有较好的疗效。
