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目的在大肠杆菌中表达和纯化一种新型的人ＩＦＮα１ｃ。方法首先建立了表达和纯化ＩＦＮα１ｃ的实验室生产流程。根据干扰素的抗病毒、抗细胞增殖以及免疫调节特性，体外测定这种ＩＦＮ的生物学活性。结果ＥＬＩＳＡ和Ｗｅｓｔｅｒｎ免疫印迹均证实，表达产物具有干扰素的免疫反应性。ＶＳＶ－ＷＩＳＨ系统的抗病毒测活表明，表达的 ＩＦＮα１ｃ（３． ２ × １０７）的抗病毒活性较 ＩＦＮα１ｂ（１． ０ × １０７－ １． ８ × １０７）略高；而抗 Ａ５４９细胞的增殖能力则与后者相当。此外ＩＦＮα１ｃ还能增强ＮＫ细胞的杀伤活性。结论本系统成功地在大肠杆菌中表达了人ＩＦＮα１ｃ。这种高比活性的干扰素可能成为临床治疗的侯选者。
Objective To express and purify a novel human IFNα1c in Escherichia coli. Methods Firstly, the laboratory production process of expressing and purifying IFNα1c was established. The biological activity of this IFN was determined in vitro based on the antiviral, anti-cellular proliferation and immunomodulatory properties of the interferon. Results Both ELISA and Western immunoblotting confirmed that the expressed product had the immunoreactivity of interferon. The antiviral assay of VSV-WISH system showed that the antiviral activity of IFNα1c (3.2 × 107) was slightly higher than that of IFNα1b (1.0 × 107-1.8 × 107), while the proliferation of A549 cells The same as the latter. In addition IFNα1c can also enhance NK cell killing activity. Conclusion This system successfully expressed human IFNα1c in E. coli. This high specific activity of interferon may become a candidate for clinical treatment.