摘要
白细胞介素-1与免疫球蛋白E在过敏性哮喘发病中发挥着重要作用。本试验克隆了白细胞介素-1受体拮抗剂(IL-1ra)及IgE分子恒定区cDNA片段,构建了融合基因原核表达载体IL-1ra-Fcε/pBV220。将其转化大肠杆菌BL21(DE3),实现了融合蛋白的高效表达,Western blotting结果表明表达蛋白为目的融合蛋白,主要以包涵体形式存在;利用分子筛和阳离子交换层析对表达产物经进行了纯化,纯化的包涵体复性后经体外功能试验表明,融合蛋白的活性与IL-1ra没有显著性差异;初步药代动力学分析显示IL-1ra-Fcε半衰期比IL-1ra延长了4.78倍。
Both interleukin-1 and IgE are important in the pathogenic mechanism of the allergy asthma, cDNA of interleukinlreceptor antagonist (IL-1ra) and IgE were cloned and a prokaryotic expression vector IL-1ra-Fcε/pBV220 was constructed. The vector was transformed into Escherichia coli BL21(DE3). The fusion protein was expressed successfully in the form of inclusion body. The recombination protein of IL-1ra-Fcε was highly purified by chromatography of gel filtration and ion exchange, which was identifited by Western blotting. The cell assay showed that the activity of IL-1ra-Fcε was as high as IL-1ra in vitro after refolding. The pharmacokenetic profile of IL-1ra-Fcε and IL-1ra was analyzed, and the half time of IL-1ra-Fcε is 4.78 times than that of IL-1ra.
出处
《生物工程学报》
CAS
CSCD
北大核心
2008年第10期1754-1760,共7页
Chinese Journal of Biotechnology
基金
国家科技支撑计划课题(No.2006BAI19B05-3)资助~~