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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2011-01-01
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doaj-95907b99ff4c430e832e4199633ac4c72020-11-25T00:52:36ZengPublic Library of Science (PLoS)PLoS ONE1932-62032011-01-0162e1470210.1371/journal.pone.0014702Expanding the versatility of phage display I: efficient display of peptide-tags on protein VII of the filamentous phage.Geir Åge LøsetBjarne BogenInger SandlieBACKGROUND: 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.http://europepmc.org/articles/PMC3044727?pdf=render |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Geir Åge Løset Bjarne Bogen Inger Sandlie |
spellingShingle |
Geir Åge Løset Bjarne Bogen Inger Sandlie Expanding the versatility of phage display I: efficient display of peptide-tags on protein VII of the filamentous phage. PLoS ONE |
author_facet |
Geir Åge Løset Bjarne Bogen Inger Sandlie |
author_sort |
Geir Åge Løset |
title |
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_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_sort |
expanding the versatility of phage display i: efficient display of peptide-tags on protein vii of the filamentous phage. |
publisher |
Public Library of Science (PLoS) |
series |
PLoS ONE |
issn |
1932-6203 |
publishDate |
2011-01-01 |
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. |
url |
http://europepmc.org/articles/PMC3044727?pdf=render |
work_keys_str_mv |
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