PURPOSE: A construction method of a phage display library using helper phage variants is provided. It can be useful for developing antibodies specific to a certain antigen and polypeptide ligands binding with protein or polypeptide. CONSTITUTION: The helper phage variants for packaging phagemid vector containing filamentous bacteriophage genome, in which the total or a portion of the wild minor coat protein gene is deleted or incomplete, contains the wild minor coat protein gene in its genome in which the gene encoding N-terminal region of minor coat protein contains an optionally suppressed translation termination codon. The filamentous bacteriophage is selected from fd, M13, f1, If1, Ike, Zj/Z, Ff, Xf, Pf1 and Pf3; the minor coat protein is pIII. The optionally suppressed translation termination codon is UAG(Amber), UAA(Ocher) or UGA(Opel) codon. The construction method of the phage display library using helper phage variants comprises the steps of: (1) preparing a recombinant phagemid expressing a heterogous protein fused with an anchor domain of pIII; (2) co-infecting the prepared phagemid with the helper phage variants to a suppressing host cell; and (3) culturing the host cell to produce a recombinant virus expressing a fusion protein of pIII and heterogous protein.
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