Journal Article

DeltaPhage—a novel helper phage for high-valence pIX phagemid display

Nicolay R. Nilssen, Terje Frigstad, Sylvie Pollmann, Norbert Roos, Bjarne Bogen, Inger Sandlie and Geir Å. Løset

in Nucleic Acids Research

Volume 40, issue 16, pages e120-e120
Published in print September 2012 | ISSN: 0305-1048
Published online April 2012 | e-ISSN: 1362-4962 | DOI: http://dx.doi.org/10.1093/nar/gks341

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Phage display has been instrumental in discovery of novel binding peptides and folded domains for the past two decades. We recently reported a novel pIX phagemid display system that is characterized by a strong preference for phagemid packaging combined with low display levels, two key features that support highly efficient affinity selection. However, high diversity in selected repertoires are intimately coupled to high display levels during initial selection rounds. To incorporate this additional feature into the pIX display system, we have developed a novel helper phage termed DeltaPhage that allows for high-valence display on pIX. This was obtained by inserting two amber mutations close to the pIX start codon, but after the pVII translational stop, conditionally inactivating the helper phage encoded pIX. Until now, the general notion has been that display on pIX is dependent on wild-type complementation, making high-valence display unachievable. However, we found that DeltaPhage does facilitate high-valence pIX display when used with a non-suppressor host. Here, we report a side-by-side comparison with pIII display, and we find that this novel helper phage complements existing pIX phagemid display systems to allow both low and high-valence display, making pIX display a complete and efficient alternative to existing pIII phagemid display systems.

Journal Article.  7832 words.  Illustrated.

Subjects: Chemistry ; Biochemistry ; Bioinformatics and Computational Biology ; Genetics and Genomics ; Molecular and Cell Biology

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