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

Abstract

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 combinationmore » of phage and yeast display to select intriguing peptide ligands. Here, 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.« less

Authors:
; ; ; ; ; ; ; ; ; ;
Publication Date:
Research Org.:
Los Alamos National Lab. (LANL), Los Alamos, NM (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES)
OSTI Identifier:
1337465
Alternate Identifier(s):
OSTI ID: 1325648
Report Number(s):
LA-UR-16-21158
Journal ID: ISSN 1932-6203; 10.1371/journal.pone.0160940
Grant/Contract Number:  
AC52-06NA25396
Resource Type:
Published Article
Journal Name:
PLoS ONE
Additional Journal Information:
Journal Name: PLoS ONE Journal Volume: 11 Journal Issue: 9; Journal ID: ISSN 1932-6203
Publisher:
Public Library of Science (PLoS)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; Biological Science; Phage display

Citation Formats

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., and Goldman, ed., Ellen R. Beyond Helper Phage: Using "Helper Cells" to Select Peptide Affinity Ligands. United States: N. p., 2016. Web. doi:10.1371/journal.pone.0160940.
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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., & Goldman, ed., Ellen R. Beyond Helper Phage: Using "Helper Cells" to Select Peptide Affinity Ligands. United States. https://doi.org/10.1371/journal.pone.0160940
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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., and Goldman, ed., Ellen R. Wed . "Beyond Helper Phage: Using "Helper Cells" to Select Peptide Affinity Ligands". United States. https://doi.org/10.1371/journal.pone.0160940.
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@article{osti_1337465,
title = {Beyond Helper Phage: Using "Helper Cells" to Select Peptide Affinity Ligands},
author = {Phipps, M. Lisa and Lillo, Antoinetta M. and Shou, Yulin and Schmidt, Emily N. and Paavola, Chad D. and Naranjo, Leslie and Bemdich, Sara and Swanson, Basil I. and Bradbury, Andrew R. M. and Martinez, Jennifer S. and Goldman, ed., Ellen R.},
abstractNote = {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. Here, 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.},
doi = {10.1371/journal.pone.0160940},
journal = {PLoS ONE},
number = 9,
volume = 11,
place = {United States},
year = {2016},
month = {9}
}
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https://doi.org/10.1371/journal.pone.0160940
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    Selection of phage-displayed accessible recombinant targeted antibodies (SPARTA): methodology and applications
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