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Expanding the Versatility of Phage Display I: Efficient Display of Peptide-Tags on Protein VII of the Filamentous Phage

BACKGROUND: Phage display is a platform for selection of specific binding molecules and this is a clear-cut motivation for increasing its performance. Polypeptides are normally displayed as fusions to the major coat protein VIII (pVIII), or the minor coat protein III (pIII). Display on other coat pr...

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Detalles Bibliográficos
Autores principales: Løset, Geir Åge, Bogen, Bjarne, Sandlie, Inger
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3044727/
https://www.ncbi.nlm.nih.gov/pubmed/21390217
http://dx.doi.org/10.1371/journal.pone.0014702
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author Løset, Geir Åge
Bogen, Bjarne
Sandlie, Inger
author_facet Løset, Geir Åge
Bogen, Bjarne
Sandlie, Inger
author_sort Løset, Geir Åge
collection PubMed
description BACKGROUND: Phage display is a platform for selection of specific binding molecules and this is a clear-cut motivation for increasing its performance. Polypeptides are normally displayed as fusions to the major coat protein VIII (pVIII), or the minor coat protein III (pIII). Display on other coat proteins such as pVII allows for display of heterologous peptide sequences on the virions in addition to those displayed on pIII and pVIII. In addition, pVII display is an alternative to pIII or pVIII display. METHODOLOGY/PRINCIPAL FINDINGS: Here we demonstrate how standard pIII or pVIII display phagemids are complemented with a helper phage which supports production of virions that are tagged with octa FLAG, HIS(6) or AviTag on pVII. The periplasmic signal sequence required for pIII and pVIII display, and which has been added to pVII in earlier studies, is omitted altogether. CONCLUSIONS/SIGNIFICANCE: Tagging on pVII is an important and very useful add-on feature to standard pIII and pVII display. Any phagemid bearing a protein of interest on either pIII or pVIII can be tagged with any of the tags depending simply on choice of helper phage. We show in this paper how such tags may be utilized for immobilization and separation as well as purification and detection of monoclonal and polyclonal phage populations.
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spelling pubmed-30447272011-03-09 Expanding the Versatility of Phage Display I: Efficient Display of Peptide-Tags on Protein VII of the Filamentous Phage Løset, Geir Åge Bogen, Bjarne Sandlie, Inger PLoS One Research Article BACKGROUND: Phage display is a platform for selection of specific binding molecules and this is a clear-cut motivation for increasing its performance. Polypeptides are normally displayed as fusions to the major coat protein VIII (pVIII), or the minor coat protein III (pIII). Display on other coat proteins such as pVII allows for display of heterologous peptide sequences on the virions in addition to those displayed on pIII and pVIII. In addition, pVII display is an alternative to pIII or pVIII display. METHODOLOGY/PRINCIPAL FINDINGS: Here we demonstrate how standard pIII or pVIII display phagemids are complemented with a helper phage which supports production of virions that are tagged with octa FLAG, HIS(6) or AviTag on pVII. The periplasmic signal sequence required for pIII and pVIII display, and which has been added to pVII in earlier studies, is omitted altogether. CONCLUSIONS/SIGNIFICANCE: Tagging on pVII is an important and very useful add-on feature to standard pIII and pVII display. Any phagemid bearing a protein of interest on either pIII or pVIII can be tagged with any of the tags depending simply on choice of helper phage. We show in this paper how such tags may be utilized for immobilization and separation as well as purification and detection of monoclonal and polyclonal phage populations. Public Library of Science 2011-02-24 /pmc/articles/PMC3044727/ /pubmed/21390217 http://dx.doi.org/10.1371/journal.pone.0014702 Text en Løset et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Løset, Geir Åge
Bogen, Bjarne
Sandlie, Inger
Expanding the Versatility of Phage Display I: Efficient Display of Peptide-Tags on Protein VII of the Filamentous Phage
title Expanding the Versatility of Phage Display I: Efficient Display of Peptide-Tags on Protein VII of the Filamentous Phage
title_full Expanding the Versatility of Phage Display I: Efficient Display of Peptide-Tags on Protein VII of the Filamentous Phage
title_fullStr Expanding the Versatility of Phage Display I: Efficient Display of Peptide-Tags on Protein VII of the Filamentous Phage
title_full_unstemmed Expanding the Versatility of Phage Display I: Efficient Display of Peptide-Tags on Protein VII of the Filamentous Phage
title_short Expanding the Versatility of Phage Display I: Efficient Display of Peptide-Tags on Protein VII of the Filamentous Phage
title_sort expanding the versatility of phage display i: efficient display of peptide-tags on protein vii of the filamentous phage
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3044727/
https://www.ncbi.nlm.nih.gov/pubmed/21390217
http://dx.doi.org/10.1371/journal.pone.0014702
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