title: Tresyl-based conjugation of protein antigen to lipid nanoparticles increases antigen immunogenicity.
abstract: The present studies were aimed at investigating the engineering of NPs with protein-conjugated-surfactant at their surface. In order to increase the immunogenicity of a protein antigen, Brij 78 was functionalized by tresyl chloride and then further reacted with the primary amine of the model proteins ovalbumin (OVA) or horseradish peroxide (HRP). The reaction yielded Brij 78-OVA and Brij 78-HRP conjugates which were then used directly to form NP-OVA or NP-HRP using a one-step warm oil-in-water microemulsion precursor method with emulsifying wax as the oil phase, and Brij 78 and the Brij 78-OVA or Brij 78-HRP conjugate as surfactants. Similarly, Brij 700 was conjugated to HIV p24 antigen to yield Brij 700-p24 conjugate. The utility of these NPs for enhancing the immune responses to protein-based vaccines was evaluated in vivo using ovalbumin (OVA) as model protein and p24 as a relevant HIV antigen. In separate in vivo studies, female BALB/c mice were immunized by subcutaneous (s.c.) injection with NP-OVA and NP-p24 formulations along with several control formulations. These results suggested that with multiple antigens, covalent attachment of the antigen to the NP significantly enhanced antigen-specific immune responses. This facile covalent conjugation and incorporation method may be utilized to further incorporate other protein antigens, even multiple antigens, into an enhanced vaccine delivery system.
authors.affiliation: Division of Molecular Pharmaceutics, and the Center for Nanotechnology in Drug Delivery, UNC Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, NC 27514, United States.
authors.forename: Anekant
authors.initials: A
authors.lastname: Jain
authors.forename: Weili
authors.initials: W
authors.lastname: Yan
authors.forename: Keith R
authors.initials: KR
authors.lastname: Miller
authors.forename: Ronan
authors.initials: R
authors.lastname: O'Carra
authors.forename: Jerold G
authors.initials: JG
authors.lastname: Woodward
authors.forename: Russell J
authors.initials: RJ
authors.lastname: Mumper
mesh_terms: D000276:Adjuvants, Immunologic / Q000737:chemistry; D000818:Animals; D000917:Antibody Formation; D016503:Drug Delivery Systems; D005260:Female; D016655:HIV Core Protein p24 / Q000737:chemistry / Q000276:immunology; D008055:Lipids / Q000737:chemistry*; D051379:Mice; D008807:Mice, Inbred BALB C; D053758:Nanoparticles; D010047:Ovalbumin / Q000737:chemistry / Q000276:immunology; D011092:Polyethylene Glycols / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry* / Q000276:immunology; D013450:Sulfones / Q000737:chemistry*; D014612:Vaccines / Q000737:chemistry / Q000276:immunology
chemical_list: D000276:Adjuvants, Immunologic; D016655:HIV Core Protein p24; D008055:Lipids; D011506:Proteins; D013450:Sulfones; D014612:Vaccines; C032605:2,2,2-trifluoroethanesulfonyl chloride; D011092:Polyethylene Glycols; C043444:octadecyl polyoxyethylene ether; D010047:Ovalbumin