Engineering human interferon alpha1c/86D with phage display technology.
Human interferon-alpha1c/86D (IFNalpha1c/86D) was functionally displayed on the surface of the filamentous bacteriophage using a phagemid vector system (pCANTAB5E). 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-alpha1b, 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 alpha1c/86D variants with increased specific activity might be obtained after purification procedures.
Ma
X
X
State Key Laboratory for Molecular Virology and Genetic Engineering, Beijing, China.
Hu
R
R
Lü
H
H
Wei
K
K
Zhang
L
L
Xue
S
S
Hou
Y
Y