Phage display is an extraordinarily powerful and versatile technology that enables the selection of novel binding functions from large populations of randomly generated peptide sequences. From a sufficiently complex library, phage bearing peptides with practically any desired binding activity can be physically isolated by affinity selection, and, since each particle carries in its genome the genetic information for its own replication, the selectants can be amplified in bacteria.
This aim of this project is to develop a new platform for peptide display on virus-like particles (VLPs) of the RNA bacteriophage MS2. We envision several applications for the MS2 VLP, but we wish especially to emphasize its utility for vaccine development. It will integrate into a single platform the potent immunogenicity of a VLP with the affinity selection capability of conventional phage display. Filamentous phages are now the most widely used vehicles for phage display, and provide an efficient means for epitope identification. However, the peptides they display are typically poorly immunogenic, because they do not normally support the formation of dense repetitive arrays. Meanwhile, other VLP systems permit engineered display of specfic pre-selected epitopes, but are incapable of peptide library display and affinity selection. We think MS2 VLPs will overcome these limitations. Peptides displayed on MS2 VLPs are strongly immunogenic, and can be engineered to encapsidate the same mRNA molecule that encodes them, thus enabling recovery of affinity selected sequences by RT-PCR. Further, the comparative simplicity of MS2 VLP structure and assembly makes it possible to conduct the entire iterative selection/amplification process in vitro. This could make it easier to achieve high library complexities, and should make automation possible.

Public Health Relevance

This project aims to develop a new platform for peptide display using virus-like particles of bacteriophage MS2. Several applications are envisioned, but because of their potent immunogenicity, these particles should be especially useful for vaccine discovery.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM042901-18
Application #
7858296
Study Section
Special Emphasis Panel (ZRG1-GGG-J (10))
Program Officer
Jones, Warren
Project Start
1991-01-01
Project End
2012-05-31
Budget Start
2010-06-01
Budget End
2011-05-31
Support Year
18
Fiscal Year
2010
Total Cost
$297,000
Indirect Cost
Name
University of New Mexico
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
868853094
City
Albuquerque
State
NM
Country
United States
Zip Code
87131
Lino, Christopher A; Caldeira, Jerri C; Peabody, David S (2017) Display of single-chain variable fragments on bacteriophage MS2 virus-like particles. J Nanobiotechnology 15:13
Frietze, Kathryn M; Peabody, David S; Chackerian, Bryce (2016) Engineering virus-like particles as vaccine platforms. Curr Opin Virol 18:44-9
O'Rourke, John P; Peabody, David S; Chackerian, Bryce (2015) Affinity selection of epitope-based vaccines using a bacteriophage virus-like particle platform. Curr Opin Virol 11:76-82
Caldeira, Jerri; Bustos, Jeremiah; Peabody, Julianne et al. (2015) Epitope-Specific Anti-hCG Vaccines on a Virus Like Particle Platform. PLoS One 10:e0141407
Ashley, Carlee E; Carnes, Eric C; Phillips, Genevieve K et al. (2011) Cell-specific delivery of diverse cargos by bacteriophage MS2 virus-like particles. ACS Nano 5:5729-45
Caldeira, Jerri C; Peabody, David S (2011) Thermal stability of RNA phage virus-like particles displaying foreign peptides. J Nanobiotechnology 9:22
Chackerian, Bryce; Caldeira, Jerri do Carmo; Peabody, Julianne et al. (2011) Peptide epitope identification by affinity selection on bacteriophage MS2 virus-like particles. J Mol Biol 409:225-37
Caldeira, Jerri do Carmo; Medford, Alexander; Kines, Rhonda C et al. (2010) Immunogenic display of diverse peptides, including a broadly cross-type neutralizing human papillomavirus L2 epitope, on virus-like particles of the RNA bacteriophage PP7. Vaccine 28:4384-93
Peabody, David S; Manifold-Wheeler, Brett; Medford, Alexander et al. (2008) Immunogenic display of diverse peptides on virus-like particles of RNA phage MS2. J Mol Biol 380:252-63
Ni, C Z; White, C A; Mitchell, R S et al. (1996) Crystal structure of the coat protein from the GA bacteriophage: model of the unassembled dimer. Protein Sci 5:2485-93

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