title: A generic method for design of oligomer-specific antibodies.
abstract: Antibodies that preferentially and specifically target pathological oligomeric protein and peptide assemblies, as opposed to their monomeric and amyloid counterparts, provide therapeutic and diagnostic opportunities for protein misfolding diseases. Unfortunately, the molecular properties associated with oligomer-specific antibodies are not well understood, and this limits targeted design and development. We present here a generic method that enables the design and optimisation of oligomer-specific antibodies. The method takes a two-step approach where discrimination between oligomers and fibrils is first accomplished through identification of cryptic epitopes exclusively buried within the structure of the fibrillar form. The second step discriminates between monomers and oligomers based on differences in avidity. We show here that a simple divalent mode of interaction, as within e.g. the IgG isotype, can increase the binding strength of the antibody up to 1500 times compared to its monovalent counterpart. We expose how the ability to bind oligomers is affected by the monovalent affinity and the turnover rate of the binding and, importantly, also how oligomer specificity is only valid within a specific concentration range. We provide an example of the method by creating and characterising a spectrum of different monoclonal antibodies against both the Aβ peptide and α-synuclein that are associated with Alzheimer's and Parkinson's diseases, respectively. The approach is however generic, does not require identification of oligomer-specific architectures, and is, in essence, applicable to all polypeptides that form oligomeric and fibrillar assemblies.
authors.affiliation: Department of Medical Biochemistry and Biophysics, Umeå University, Umeå, Sweden.
authors.forename: Kristoffer
authors.initials: K
authors.lastname: Brännström
authors.affiliation: Department of Medical Biochemistry and Biophysics, Umeå University, Umeå, Sweden.
authors.forename: Malin
authors.initials: M
authors.lastname: Lindhagen-Persson
authors.affiliation: Department of Medical Biochemistry and Biophysics, Umeå University, Umeå, Sweden.
authors.forename: Anna L
authors.initials: AL
authors.lastname: Gharibyan
authors.affiliation: Department of Medical Biochemistry and Biophysics, Umeå University, Umeå, Sweden.
authors.forename: Irina
authors.initials: I
authors.lastname: Iakovleva
authors.affiliation: Department of Medical Biochemistry and Biophysics, Umeå University, Umeå, Sweden.
authors.forename: Monika
authors.initials: M
authors.lastname: Vestling
authors.affiliation: Department of Molecular Biology, Umeå University, Umeå, Sweden.
authors.forename: Mikael E
authors.initials: ME
authors.lastname: Sellin
authors.affiliation: Department of Medical Biosciences, Umeå University, Umeå, Sweden.
authors.forename: Thomas
authors.initials: T
authors.lastname: Brännström
authors.affiliation: Department of Medical Biochemistry and Biophysics, Umeå University, Umeå, Sweden.
authors.forename: Ludmilla
authors.initials: L
authors.lastname: Morozova-Roche
authors.affiliation: Department of Clinical Pharmacology and Clinical Neuroscience, Umeå University, Umeå, Sweden.
authors.forename: Lars
authors.initials: L
authors.lastname: Forsgren
authors.affiliation: Department of Medical Biochemistry and Biophysics, Umeå University, Umeå, Sweden.
authors.forename: Anders
authors.initials: A
authors.lastname: Olofsson
mesh_terms: D000544:Alzheimer Disease / Q000378:metabolism / Q000473:pathology; D016229:Amyloid beta-Peptides / Q000737:chemistry / Q000276:immunology; D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology; D000915:Antibody Affinity / Q000276:immunology; D000939:Epitopes / Q000737:chemistry / Q000276:immunology*; D005260:Female; D006624:Hippocampus / Q000378:metabolism / Q000473:pathology; D007700:Kinetics; D051379:Mice; D010300:Parkinson Disease / Q000378:metabolism / Q000473:pathology; D011485:Protein Binding; D055503:Protein Multimerization / Q000276:immunology*; D011506:Proteins / Q000737:chemistry / Q000276:immunology*; D051844:alpha-Synuclein / Q000737:chemistry / Q000276:immunology / Q000378:metabolism
chemical_list: D016229:Amyloid beta-Peptides; D000906:Antibodies; D000911:Antibodies, Monoclonal; D000939:Epitopes; D011506:Proteins; D051844:alpha-Synuclein