title: Structure-based design of conformation- and sequence-specific antibodies against amyloid β.
abstract: Conformation-specific antibodies that recognize aggregated proteins associated with several conformational disorders (e.g., Parkinson and prion diseases) are invaluable for diagnostic and therapeutic applications. However, no systematic strategy exists for generating conformation-specific antibodies that target linear sequence epitopes within misfolded proteins. Here we report a strategy for designing conformation- and sequence-specific antibodies against misfolded proteins that is inspired by the molecular interactions governing protein aggregation. We find that grafting small amyloidogenic peptides (6-10 residues) from the Aβ42 peptide associated with Alzheimer's disease into the complementarity determining regions of a domain (V(H)) antibody generates antibody variants that recognize Aβ soluble oligomers and amyloid fibrils with nanomolar affinity. We refer to these antibodies as gammabodies for grafted amyloid-motif antibodies. Gammabodies displaying the central amyloidogenic Aβ motif (18VFFA21) are reactive with Aβ fibrils, whereas those displaying the amyloidogenic C terminus (34LMVGGVVIA42) are reactive with Aβ fibrils and oligomers (and weakly reactive with Aβ monomers). Importantly, we find that the grafted motifs target the corresponding peptide segments within misfolded Aβ conformers. Aβ gammabodies fail to cross-react with other amyloidogenic proteins and scrambling their grafted sequences eliminates antibody reactivity. Finally, gammabodies that recognize Aβ soluble oligomers and fibrils also neutralize the toxicity of each Aβ conformer. We expect that our antibody design strategy is not limited to Aβ and can be used to readily generate gammabodies against other toxic misfolded proteins.
authors.affiliation: Center for Biotechnology and Interdisciplinary Studies, Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, Troy, NY 12180, USA.
authors.forename: Joseph M
authors.initials: JM
authors.lastname: Perchiacca
authors.forename: Ali Reza A
authors.initials: AR
authors.lastname: Ladiwala
authors.forename: Moumita
authors.initials: M
authors.lastname: Bhattacharya
authors.forename: Peter M
authors.initials: PM
authors.lastname: Tessier
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000682:Amyloid / Q000737:chemistry / Q000276:immunology; D016229:Amyloid beta-Peptides / Q000737:chemistry* / Q000276:immunology* / Q000633:toxicity; D000818:Animals; D000906:Antibodies / Q000737:chemistry* / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology; D000918:Antibody Specificity; D022801:Complementarity Determining Regions / Q000737:chemistry; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D016716:PC12 Cells; D011485:Protein Binding; D015202:Protein Engineering / Q000379:methods*; D020033:Protein Isoforms / Q000737:chemistry / Q000276:immunology; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D051381:Rats
chemical_list: D000682:Amyloid; D016229:Amyloid beta-Peptides; D000906:Antibodies; D000911:Antibodies, Monoclonal; D022801:Complementarity Determining Regions; D020033:Protein Isoforms