title: Multivalent pIX phage display selects for distinct and improved antibody properties.
abstract: Phage display screening readily allows for the identification of a multitude of antibody specificities, but to identify optimal lead candidates remains a challenge. Here, we direct the antibody-capsid fusion away from the signal sequence-dependent secretory SEC pathway in E. coli by utilizing the intrinsic signal sequence-independent property of pIX to obtain virion integration. This approach was combined with the use of an engineered helper phage known to improve antibody pIX display and retrieval. By direct comparison with pIII display, we demonstrate that antibody display using this pIX system translates into substantially improved retrieval of desired specificities with favorable biophysical properties in de novo selection. We show that the effect was due to less E. coli host toxicity during phage propagation conferred by the lack of a signal sequence. This pIX combinatorial display platform provides a generic alternative route for obtaining good binders with high stability and may thus find broad applicability.
authors.affiliation: Centre for Immune Regulation and Department of Immunology, University of Oslo and Oslo University Hospital, N-0372 Oslo, Norway.|Department of Biosciences, University of Oslo, N-0316 Oslo, Norway.
authors.forename: Lene S
authors.initials: LS
authors.lastname: Høydahl
authors.affiliation: Centre for Immune Regulation and Department of Immunology, University of Oslo and Oslo University Hospital, N-0372 Oslo, Norway.|Department of Biosciences, University of Oslo, N-0316 Oslo, Norway.
authors.forename: Nicolay R
authors.initials: NR
authors.lastname: Nilssen
authors.affiliation: Centre for Immune Regulation and Department of Immunology, University of Oslo and Oslo University Hospital, N-0372 Oslo, Norway.
authors.forename: Kristin S
authors.initials: KS
authors.lastname: Gunnarsen
authors.affiliation: Centre for Immune Regulation and Department of Immunology, University of Oslo and Oslo University Hospital, N-0372 Oslo, Norway.
authors.forename: M Fleur du
authors.initials: MF
authors.lastname: Pré
authors.affiliation: Centre for Immune Regulation and Department of Immunology, University of Oslo and Oslo University Hospital, N-0372 Oslo, Norway.
authors.forename: Rasmus
authors.initials: R
authors.lastname: Iversen
authors.affiliation: Department of Biosciences, University of Oslo, N-0316 Oslo, Norway.
authors.forename: Norbert
authors.initials: N
authors.lastname: Roos
authors.affiliation: Centre for Immune Regulation and Department of Immunology, University of Oslo and Oslo University Hospital, N-0372 Oslo, Norway.
authors.forename: Xi
authors.initials: X
authors.lastname: Chen
authors.affiliation: Department of Immunology, Norwegian Institute of Public Health, N-0403 Oslo, Norway.|School of Pharmacy, University of Oslo, N-0316 Oslo, Norway.
authors.forename: Terje E
authors.initials: TE
authors.lastname: Michaelsen
authors.affiliation: Centre for Immune Regulation and Department of Immunology, University of Oslo and Oslo University Hospital, N-0372 Oslo, Norway.|KG Jebsen Coeliac Disease Research Centre and Department of Immunology, University of Oslo, N-0372 Oslo, Norway.
authors.forename: Ludvig M
authors.initials: LM
authors.lastname: Sollid
authors.affiliation: Centre for Immune Regulation and Department of Immunology, University of Oslo and Oslo University Hospital, N-0372 Oslo, Norway.|Department of Biosciences, University of Oslo, N-0316 Oslo, Norway.
authors.forename: Inger
authors.initials: I
authors.lastname: Sandlie
authors.affiliation: Centre for Immune Regulation and Department of Immunology, University of Oslo and Oslo University Hospital, N-0372 Oslo, Norway.|Department of Biosciences, University of Oslo, N-0316 Oslo, Norway.|Nextera AS, N-0349 Oslo, Norway.
authors.forename: Geir Å
authors.initials: GÅ
authors.lastname: Løset
mesh_terms: D000906:Antibodies / Q000378:metabolism*; D000918:Antibody Specificity; D001435:Bacteriophages / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D036022:Capsid Proteins / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000821:virology*; D006378:Helper Viruses / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D019151:Peptide Library; D021382:Protein Sorting Signals; D014771:Virion / Q000235:genetics / Q000378:metabolism / Q000502:physiology
chemical_list: D000906:Antibodies; D036022:Capsid Proteins; D019151:Peptide Library; D021382:Protein Sorting Signals