In vitro binding of anthrax protective antigen on bacteriophage T4 capsid surface through Hoc-capsid interactions: A strategy for efficient display of large full-length proteins
- Department of Biology, 103 McCort Ward Hall, Catholic University of America, 620 Michigan Ave., NE, Washington, DC 20064 (United States)
- Division of Retrovirology, Walter Reed Army Institute of Research, 503 Robert Grant Avenue, Silver Spring, MD 20910 (United States)
- Bacterial Toxins and Therapeutics Section, National Institute of Allergy and Infectious Diseases, NIH, 30 Convent Dr., Bethesda, MD 20892 (United States)
An in vitro binding system is described to display large full-length proteins on bacteriophage T4 capsid surface at high density. The phage T4 icosahedral capsid features 155 copies of a nonessential highly antigenic outer capsid protein, Hoc, at the center of each major capsid protein hexon. Gene fusions were engineered to express the 83-kDa protective antigen (PA) from Bacillus anthracis fused to the N-terminus of Hoc and the 130-kDa PA-Hoc protein was expressed in Escherichia coli and purified. The purified PA-Hoc was assembled in vitro on hoc {sup -} phage particles. Binding was specific, stable, and of high affinity. This defined in vitro system allowed manipulation of the copy number of displayed PA and imposed no significant limitation on the size of the displayed antigen. In contrast to in vivo display systems, the in vitro approach allows all the capsid binding sites to be occupied by the 130-kDa PA-Hoc fusion protein. The PA-T4 particles were immunogenic in mice in the absence of an adjuvant, eliciting strong PA-specific antibodies and anthrax lethal toxin neutralizing antibodies. The in vitro display on phage T4 offers a novel platform for potential construction of customized vaccines against anthrax and other infectious diseases.
- OSTI ID:
- 20779456
- Journal Information:
- Virology, Journal Name: Virology Journal Issue: 1 Vol. 345; ISSN VIRLAX; ISSN 0042-6822
- Country of Publication:
- United States
- Language:
- English
Similar Records
Cryo-electron microscopy study of bacteriophage T4 displaying anthrax toxin proteins
Structure of the Three N-Terminal Immunoglobulin Domains of the Highly Immunogenic Outer Capsid Protein from a T4-Like Bacteriophage
Evidence that bacteriophage T4 ephl is a missense hoc mutation
Journal Article
·
Thu Oct 25 00:00:00 EDT 2007
· Virology
·
OSTI ID:21077985
Structure of the Three N-Terminal Immunoglobulin Domains of the Highly Immunogenic Outer Capsid Protein from a T4-Like Bacteriophage
Journal Article
·
Fri Sep 16 00:00:00 EDT 2011
· J. Virol.
·
OSTI ID:1021776
Evidence that bacteriophage T4 ephl is a missense hoc mutation
Journal Article
·
Sun May 01 00:00:00 EDT 1983
· J. Virol.; (United States)
·
OSTI ID:6471112