Engineering human interferon alpha 1c/86D with phage display technology

作者：Ma, XJ^*; Hu, R; Lu, H; Wei, KK; Zhang, LL; Xue, SX; Hou, YD

来源：Science China Life Sciences, 1999, 42(2): 191-201.

DOI：10.1007/BF02880056

• phage display
• peptide library
• human interferon alpha 1

Human interferon-alpha 1c/86D (IFN alpha 1c/86D) was functionally displayed on the surface of the filamentous bacteriophage using a phagemid vector system (pCANTABSE). The key amino acid residues involved in the receptor binding were further defined with phage displayed 6-mer peptide library and two neutralizing antibodies against linear epitopes on the IFN-alpha lb, indicating that residues 30, 33, 34, (AB-loop) and residues 124, 126, 127 (D helix, DE-loop) were more critical than the adjacent residues for recognition of receptor. In addition, a cassette mutagenesis library was generated by fully randomizing the sequence of the four positions 29, 31, 32 and 35 in AB-loop, and used to select phage-IFN variants with WISH-based panning method. Three phage-IFN variants were isolated to possess more antiviral activity in the range of 4-16-fold than parental phage-IFN after IPTG-induced soluble expression. The results suggest that phage displayed phage-IFN alpha 1c/86D Variants with increased specific activity might be obtained after purification procedures.

• 出版日期1999-4
• 单位大连医科大学