Development and biological activity of long-acting recombinant human interferon-α2b

摘要

Construction and detection of expression vectors, validation of target proteins, and assessment of cell strain stability. Schematic representation of expression vectors IFN-1CTPON and IFN-2CTPON. SP, Signal peptide; N, N-glycosylation sequence. Restriction mapping confirmed the construction of the expression vectors. M, DNA size marker; lane 1, IFN-1CTPON DNA fragment inserted into the digested pEE12.4 plasmid; lane 2, IFN-2CTPON DNA fragment inserted into the digested pEE12.4 plasmid; lane 3, expression vector pEE12.4-IFN-1CTPON doubly digested with dIII and RI; lane 4, expression vector pEE12.4-IFN-2CTPON doubly digested with dIII and RI. Western blot of culture supernatants containing IFN-CTPON. M, protein molecular size marker; lane 1, culture supernatant containing IFN-1CTPON; lane 2, culture supernatant containing IFN-2CTPON. d Monoclonal CHO cell stability verified with PCR. M, DNA size marker; lane 1, IFN-1CTPON (642 bp); lane 2, IFN-2CTPON (713 bp)