title: Generation and characterization of anti-Adeno-associated virus serotype 8 (AAV8) and anti-AAV9 monoclonal antibodies.
abstract: Adeno-associated viruses (AAVs) are promising viral vectors for therapeutic gene delivery, and the approval of an AAV1 vector for the treatment of lipoprotein lipase deficiency has heralded a new and exciting era for this system. However, preclinical and clinical studies show that neutralization from pre-existing antibodies is detrimental for medical application and this hurdle must be overcome before full clinical realization can be achieved. Thus the binding sites for capsid antibodies must be identified and eliminated through capsid engineering. Towards this goal and to recapitulate patient polyclonal responses, a panel of six new mouse monoclonal antibodies (MAbs) has been generated against AAV8 and AAV9 capsids, two vectors being developed for therapeutic application. Native (capsid) dot blot assays confirmed the specificity of these antibodies for their parental serotypes, with the exception of one MAb, HL2372, selected to cross-react against both capsids. Furthermore, in vitro assays showed that these MAbs are capable of neutralizing virus infection. These MAbs will be utilized for structural mapping of antigenic footprints on their respective capsids to inform development of the next generation of rAAV vectors capable of evading antibody neutralization while retaining parental tropism.
authors.affiliation: Department of Biochemistry and Molecular Biology, Center for Structural Biology, McKnight Brain Institute, College of Medicine, University of Florida, Gainesville, FL, USA.
authors.forename: Yu-Shan
authors.initials: YS
authors.lastname: Tseng
authors.affiliation: Department of Biochemistry and Molecular Biology, Center for Structural Biology, McKnight Brain Institute, College of Medicine, University of Florida, Gainesville, FL, USA.
authors.forename: Kim Van
authors.initials: KV
authors.lastname: Vliet
authors.affiliation: Department of Genetics and The Gene Therapy Center, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
authors.forename: Lavanya
authors.initials: L
authors.lastname: Rao
authors.affiliation: Department of Biochemistry and Molecular Biology, Center for Structural Biology, McKnight Brain Institute, College of Medicine, University of Florida, Gainesville, FL, USA.
authors.forename: Robert
authors.initials: R
authors.lastname: McKenna
authors.affiliation: Department Molecular Genetics and Microbiology, and Powell Gene Therapy Center, College of Medicine, University of Florida, Gainesville, FL, USA.
authors.forename: Barry J
authors.initials: BJ
authors.lastname: Byrne
authors.affiliation: Department of Genetics and The Gene Therapy Center, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
authors.forename: Aravind
authors.initials: A
authors.lastname: Asokan
authors.affiliation: Department of Biochemistry and Molecular Biology, Center for Structural Biology, McKnight Brain Institute, College of Medicine, University of Florida, Gainesville, FL, USA. Electronic address: mckenna@ufl.edu.
authors.forename: Mavis
authors.initials: M
authors.lastname: Agbandje-McKenna
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification*; D000914:Antibodies, Viral / Q000276:immunology* / Q000302:isolation & purification*; D000229:Dependovirus / Q000276:immunology*; D006801:Humans; D051379:Mice; D065288:Serogroup
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral
keywords: AAV8; AAV9; Adeno-associated viruses; Anti-AAV capsid antibodies; Host-immune escape