Cargando…
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...
Autores principales: | , , |
---|---|
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 |
_version_ | 1782198766238957568 |
---|---|
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. |
format | Text |
id | pubmed-3044727 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT løsetgeirage expandingtheversatilityofphagedisplayiefficientdisplayofpeptidetagsonproteinviiofthefilamentousphage AT bogenbjarne expandingtheversatilityofphagedisplayiefficientdisplayofpeptidetagsonproteinviiofthefilamentousphage AT sandlieinger expandingtheversatilityofphagedisplayiefficientdisplayofpeptidetagsonproteinviiofthefilamentousphage |