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利用噬菌体表面显示技术表达具生物活性的干扰素α1c,并通过构建和筛选突变文库 ,以得到高受体结合能力和高比活性的新α1c干扰素 .利用噬菌粒载体pCANTAB5E表达了人IFNα1c/ 86D基因 ,利用随机 6肽库和干扰素的中和抗体推导干扰素α1c的受体结合区及关键性结合残基 ,运用盒式突变构建了基于AB环的突变库 ,其中氨基酸 2 9,3 1,3 2 ,3 5位完全随机化 ,并直接利用WISH细胞选择出高抗病毒活性的干扰素突变体 .结果获得了 3个重组噬菌体干扰素突变体 ,其抗病毒活性是干扰素母体的 4～ 16倍 .这一结果为国内外首次报道利用噬菌体表面显示技术的筛选表达优势改造干扰素以提高抗病毒活性 ,实验结果说明了其可行性 ,并提示干扰素突变体的比活性有可能较母体高 .
The biologically active interferon α1c was expressed by phage display technique and a new α1c interferon with high receptor binding ability and high specific activity was obtained by constructing and screening mutant library.The human IFNα1c / 86D gene was used to deduce the receptor binding region and the key binding residues of interferon α1c using neutralizing antibodies of random 6-peptide library and interferon. The AB loop-based mutant library was constructed by cassette mutation. Among them, amino acids 29.3 1,3 2 and 35 were completely randomized and WISH cells were directly used to select the high antiviral activity of interferon mutants.Results Three recombinant phage interferon mutants were obtained and their antiviral activity was interferon 4 ~ 16 times.This result is the first report at home and abroad that the use of phage display technology to improve the expression of interferon to improve the anti-viral activity, the experimental results illustrate its feasibility, and prompted the specific activity of interferon mutants may Higher than maternal.