Eliminating helper phage from phage display
Abstract
Phage display technology involves the display of proteins or peptides, as coat protein fusions, on the surface of a phage or phagemid particles. Using standard technology, helper phage are essential for the replication and assembly of phagemid particles, during library production and biopanning. We have eliminated the need to add helper phage by using ‘bacterial packaging cell lines’ that provide the same functions. These cell lines contain M13-based helper plasmids that express phage packaging proteins which assemble phagemid particles as efficiently as helper phage, but without helper phage contamination. This results in genetically pure phagemid particle preparations. Furthermore, by using constructs differing in the form of gene 3 that they contain, we have shown that the display, from a single library, can be modulated between monovalent (phagemid-like) and multivalent display (phagelike) without any further engineering. These packaging cells eliminate the use of helper phage from phagemid-based selection protocols; reducing the amount of technical preparation, facilitating automation, optimizing selections by matching display levels to diversity, and effectively using the packaged phagemid particles as means to transfer genetic information at an efficiency approaching 100%.
- Authors:
-
- Los Alamos National Lab. (LANL), Los Alamos, NM (United States)
- Publication Date:
- Research Org.:
- Los Alamos National Laboratory (LANL), Los Alamos, NM (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Biological and Environmental Research (BER). Biological Systems Science Division
- OSTI Identifier:
- 1625407
- Grant/Contract Number:
- AC52-06NA25396
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Nucleic Acids Research
- Additional Journal Information:
- Journal Volume: 34; Journal Issue: 21; Journal ID: ISSN 0305-1048
- Publisher:
- Oxford University Press
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; biochemistry & molecular biology
Citation Formats
Chasteen, L., Ayriss, J., Pavlik, P., and Bradbury, A. R. M. Eliminating helper phage from phage display. United States: N. p., 2006.
Web. doi:10.1093/nar/gkl772.
Chasteen, L., Ayriss, J., Pavlik, P., & Bradbury, A. R. M. Eliminating helper phage from phage display. United States. https://doi.org/10.1093/nar/gkl772
Chasteen, L., Ayriss, J., Pavlik, P., and Bradbury, A. R. M. Mon .
"Eliminating helper phage from phage display". United States. https://doi.org/10.1093/nar/gkl772. https://www.osti.gov/servlets/purl/1625407.
@article{osti_1625407,
title = {Eliminating helper phage from phage display},
author = {Chasteen, L. and Ayriss, J. and Pavlik, P. and Bradbury, A. R. M.},
abstractNote = {Phage display technology involves the display of proteins or peptides, as coat protein fusions, on the surface of a phage or phagemid particles. Using standard technology, helper phage are essential for the replication and assembly of phagemid particles, during library production and biopanning. We have eliminated the need to add helper phage by using ‘bacterial packaging cell lines’ that provide the same functions. These cell lines contain M13-based helper plasmids that express phage packaging proteins which assemble phagemid particles as efficiently as helper phage, but without helper phage contamination. This results in genetically pure phagemid particle preparations. Furthermore, by using constructs differing in the form of gene 3 that they contain, we have shown that the display, from a single library, can be modulated between monovalent (phagemid-like) and multivalent display (phagelike) without any further engineering. These packaging cells eliminate the use of helper phage from phagemid-based selection protocols; reducing the amount of technical preparation, facilitating automation, optimizing selections by matching display levels to diversity, and effectively using the packaged phagemid particles as means to transfer genetic information at an efficiency approaching 100%.},
doi = {10.1093/nar/gkl772},
journal = {Nucleic Acids Research},
number = 21,
volume = 34,
place = {United States},
year = {Mon Nov 06 00:00:00 EST 2006},
month = {Mon Nov 06 00:00:00 EST 2006}
}
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