title: Display of a PorA peptide from Neisseria meningitidis on the bacteriophage T4 capsid surface.
abstract: The exterior of bacteriophage T4 capsid is coated with two outer capsid proteins, Hoc (highly antigenic outer capsid protein; molecular mass, 40 kDa) and Soc (small outer capsid protein; molecular mass, 9 kDa), at symmetrical positions on the icosahedron (160 copies of Hoc and 960 copies of Soc per capsid particle). Both these proteins are nonessential for phage infectivity and viability and assemble onto the capsid surface after completion of capsid assembly. We developed a phage display system which allowed in-frame fusions of foreign DNA at a unique cloning site in the 5' end of hoc or soc. A DNA fragment corresponding to the 36-amino-acid PorA peptide from Neisseria meningitidis was cloned into the display vectors to generate fusions at the N terminus of Hoc or Soc. The PorA-Hoc and PorA-Soc fusion proteins retained the ability to bind to the capsid surface, and the bound peptide was displayed in an accessible form as shown by its reactivity with specific monoclonal antibodies in an enzyme-linked immunosorbent assay. By employing T4 genetic strategies, we show that more than one subtype-specific PorA peptide can be displayed on the capsid surface and that the peptide can also be displayed on a DNA-free empty capsid. Both the PorA-Hoc and PorA-Soc recombinant phages are highly immunogenic in mice and elicit strong antipeptide antibody titers even with a weak adjuvant such as Alhydrogel or no adjuvant at all. The data suggest that the phage T4 hoc-soc system is an attractive system for display of peptides on an icosahedral capsid surface and may emerge as a powerful system for construction of the next generation multicomponent vaccines.
authors.affiliation: Department of Biology, The Catholic University of America, Washington, D.C. 20064, USA.
authors.forename: J
authors.initials: J
authors.lastname: Jiang
authors.forename: L
authors.initials: L
authors.lastname: Abu-Shilbayeh
authors.forename: V B
authors.initials: VB
authors.lastname: Rao
mesh_terms: D000818:Animals; D001428:Bacterial Vaccines / Q000276:immunology; D017122:Bacteriophage T4 / Q000235:genetics*; D002213:Capsid / Q000378:metabolism*; D005822:Genetic Vectors; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D009345:Neisseria meningitidis / Q000276:immunology*; D018272:Porins / Q000096:biosynthesis* / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000276:immunology; D014614:Vaccines, Synthetic / Q000276:immunology
chemical_list: D001428:Bacterial Vaccines; D018272:Porins; D011993:Recombinant Fusion Proteins; D014614:Vaccines, Synthetic; C057699:porin protein, Neisseria