Eliminating Helper Phage from Phage Display.
From: B Division, Los Alamos National Laboratory MS M888, Los Alamos, NM 87545, USA.
Nucleic acids research
- Publish Date: 2006
- ISSN: 1362-4962
- Volume: 34
- Issue: 21
- Pages: e145
- Medium: Internet
- Language: English
- Citation (JAMA): Chasteen L, Ayriss J, Pavlik P, et al. Eliminating Helper Phage from Phage Display.. Nucleic Acids Res. 2006;34:e145
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 (phage-like) 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%.
Mesh Headings (Keywords): Bacteria, Bacteriophage M13, Blotting, Western, Enzyme-Linked Immunosorbent Assay, Genetic Vectors, Peptide Library, Plasmids, Viral Proteins, Virion, Virus Assembly
Check for Full Text / PubMed Unique Identifier (PMID): 17088290
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