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Beyond Helper Phage: Using "Helper Cells" to Select Peptide Affinity Ligands

Peptides are important affinity ligands for microscopy, biosensing, and targeted delivery. However, because they can have low affinity for their targets, their selection from large naïve libraries can be challenging. When selecting peptidic ligands from display libraries, it is important to: 1) ensu...

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Autores principales: Phipps, M. Lisa, Lillo, Antoinetta M., Shou, Yulin, Schmidt, Emily N., Paavola, Chad D., Naranjo, Leslie, Bemdich, Sara, Swanson, Basil I., Bradbury, Andrew R. M., Martinez, Jennifer S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5023105/
https://www.ncbi.nlm.nih.gov/pubmed/27626637
http://dx.doi.org/10.1371/journal.pone.0160940
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author Phipps, M. Lisa
Lillo, Antoinetta M.
Shou, Yulin
Schmidt, Emily N.
Paavola, Chad D.
Naranjo, Leslie
Bemdich, Sara
Swanson, Basil I.
Bradbury, Andrew R. M.
Martinez, Jennifer S.
author_facet Phipps, M. Lisa
Lillo, Antoinetta M.
Shou, Yulin
Schmidt, Emily N.
Paavola, Chad D.
Naranjo, Leslie
Bemdich, Sara
Swanson, Basil I.
Bradbury, Andrew R. M.
Martinez, Jennifer S.
author_sort Phipps, M. Lisa
collection PubMed
description Peptides are important affinity ligands for microscopy, biosensing, and targeted delivery. However, because they can have low affinity for their targets, their selection from large naïve libraries can be challenging. When selecting peptidic ligands from display libraries, it is important to: 1) ensure efficient display; 2) maximize the ability to select high affinity ligands; and 3) minimize the effect of the display context on binding. The “helper cell” packaging system has been described as a tool to produce filamentous phage particles based on phagemid constructs with varying display levels, while remaining free of helper phage contamination. Here we report on the first use of this system for peptide display, including the systematic characterization and optimization of helper cells, their inefficient use in antibody display and their use in creating and selecting from a set of phage display peptide libraries. Our libraries were analyzed with unprecedented precision by standard or deep sequencing, and shown to be superior in quality than commercial gold standards. Using our helper cell libraries, we have obtained ligands recognizing Yersinia pestis surface antigen F1V and L-glutamine-binding periplasmic protein QBP. In the latter case, unlike any of the peptide library selections described so far, we used a combination of phage and yeast display to select intriguing peptide ligands. Based on the success of our selections we believe that peptide libraries obtained with helper cells are not only suitable, but preferable to traditional phage display libraries for selection of peptidic ligands.
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spelling pubmed-50231052016-09-27 Beyond Helper Phage: Using "Helper Cells" to Select Peptide Affinity Ligands Phipps, M. Lisa Lillo, Antoinetta M. Shou, Yulin Schmidt, Emily N. Paavola, Chad D. Naranjo, Leslie Bemdich, Sara Swanson, Basil I. Bradbury, Andrew R. M. Martinez, Jennifer S. PLoS One Research Article Peptides are important affinity ligands for microscopy, biosensing, and targeted delivery. However, because they can have low affinity for their targets, their selection from large naïve libraries can be challenging. When selecting peptidic ligands from display libraries, it is important to: 1) ensure efficient display; 2) maximize the ability to select high affinity ligands; and 3) minimize the effect of the display context on binding. The “helper cell” packaging system has been described as a tool to produce filamentous phage particles based on phagemid constructs with varying display levels, while remaining free of helper phage contamination. Here we report on the first use of this system for peptide display, including the systematic characterization and optimization of helper cells, their inefficient use in antibody display and their use in creating and selecting from a set of phage display peptide libraries. Our libraries were analyzed with unprecedented precision by standard or deep sequencing, and shown to be superior in quality than commercial gold standards. Using our helper cell libraries, we have obtained ligands recognizing Yersinia pestis surface antigen F1V and L-glutamine-binding periplasmic protein QBP. In the latter case, unlike any of the peptide library selections described so far, we used a combination of phage and yeast display to select intriguing peptide ligands. Based on the success of our selections we believe that peptide libraries obtained with helper cells are not only suitable, but preferable to traditional phage display libraries for selection of peptidic ligands. Public Library of Science 2016-09-14 /pmc/articles/PMC5023105/ /pubmed/27626637 http://dx.doi.org/10.1371/journal.pone.0160940 Text en https://creativecommons.org/publicdomain/zero/1.0/ This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 (https://creativecommons.org/publicdomain/zero/1.0/) public domain dedication.
spellingShingle Research Article
Phipps, M. Lisa
Lillo, Antoinetta M.
Shou, Yulin
Schmidt, Emily N.
Paavola, Chad D.
Naranjo, Leslie
Bemdich, Sara
Swanson, Basil I.
Bradbury, Andrew R. M.
Martinez, Jennifer S.
Beyond Helper Phage: Using "Helper Cells" to Select Peptide Affinity Ligands
title Beyond Helper Phage: Using "Helper Cells" to Select Peptide Affinity Ligands
title_full Beyond Helper Phage: Using "Helper Cells" to Select Peptide Affinity Ligands
title_fullStr Beyond Helper Phage: Using "Helper Cells" to Select Peptide Affinity Ligands
title_full_unstemmed Beyond Helper Phage: Using "Helper Cells" to Select Peptide Affinity Ligands
title_short Beyond Helper Phage: Using "Helper Cells" to Select Peptide Affinity Ligands
title_sort beyond helper phage: using "helper cells" to select peptide affinity ligands
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5023105/
https://www.ncbi.nlm.nih.gov/pubmed/27626637
http://dx.doi.org/10.1371/journal.pone.0160940
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