title: Whole-Virus Screening to Develop Synbodies for the Influenza Virus.
abstract: There is an ongoing need for affinity agents for emerging viruses and new strains of current human viruses. We therefore developed a robust and modular system for engineering high-affinity synbody ligands for the influenza A/Puerto Rico/8/1934 H1N1 virus as a model system. Whole-virus screening against a peptide microarray was used to identify binding peptides. Candidate peptides were linked to bis-maleimide peptide scaffolds to produce a library of candidate influenza-binding synbodies. From this library, a candidate synbody, ASU1060, was selected and affinity-improved via positional substitution using d-amino acids to produce a new synbody, ASU1061, that bound H1N1 in an ELISA assay with a KDof <1 nM, comparable to that of a monoclonal antibody for neuraminidase (NA). We prepared a modified version of ASU1061 that contained an additional C-terminal peptide to simulate conjugation of the synbody to a carrier protein, called ASU1063, and found that H1N1 binding was unchanged. Subsequent work identified the synbody target as nucleoprotein (NP), a highly conserved protein in influenza, with a KDof <1 nM for ASU1063. This suggests that virus-binding synbodies can be conjugated to carrier proteins or other moieties that could improve the therapeutic profile of the resulting synbody. This method is a rapid process that offers a means of developing new affinity ligands to influenza and other viruses.
authors.affiliation: The Biodesign Institute Center for Innovations in Medicine, Arizona State University , Tempe, Arizona 85287, United States.
authors.forename: Nidhi
authors.initials: N
authors.lastname: Gupta
authors.affiliation: The Biodesign Institute Center for Innovations in Medicine, Arizona State University , Tempe, Arizona 85287, United States.
authors.forename: John
authors.initials: J
authors.lastname: Lainson
authors.affiliation: The Biodesign Institute Center for Innovations in Medicine, Arizona State University , Tempe, Arizona 85287, United States.
authors.forename: Valeriy
authors.initials: V
authors.lastname: Domenyuk
authors.affiliation: The Biodesign Institute Center for Innovations in Medicine, Arizona State University , Tempe, Arizona 85287, United States.
authors.forename: Zhan-Gong
authors.initials: ZG
authors.lastname: Zhao
authors.affiliation: The Biodesign Institute Center for Innovations in Medicine, Arizona State University , Tempe, Arizona 85287, United States.
authors.forename: Stephen Albert
authors.initials: SA
authors.lastname: Johnston
authors.affiliation: The Biodesign Institute Center for Innovations in Medicine, Arizona State University , Tempe, Arizona 85287, United States.
authors.forename: Chris W
authors.initials: CW
authors.lastname: Diehnelt
mesh_terms: D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D053118:Influenza A Virus, H1N1 Subtype / Q000276:immunology* / Q000378:metabolism; D009698:Nucleoproteins / Q000378:metabolism; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*
chemical_list: D009698:Nucleoproteins; D019151:Peptide Library; D010455:Peptides