title: Engineering the surface properties of a human monoclonal antibody prevents self-association and rapid clearance in vivo.
abstract: Uncontrolled self-association is a major challenge in the exploitation of proteins as therapeutics. Here we describe the development of a structural proteomics approach to identify the amino acids responsible for aberrant self-association of monoclonal antibodies and the design of a variant with reduced aggregation and increased serum persistence in vivo. We show that the human monoclonal antibody, MEDI1912, selected against nerve growth factor binds with picomolar affinity, but undergoes reversible self-association and has a poor pharmacokinetic profile in both rat and cynomolgus monkeys. Using hydrogen/deuterium exchange and cross-linking-mass spectrometry we map the residues responsible for self-association of MEDI1912 and show that disruption of the self-interaction interface by three mutations enhances its biophysical properties and serum persistence, whilst maintaining high affinity and potency. Immunohistochemistry suggests that this is achieved via reduction of non-specific tissue binding. The strategy developed represents a powerful and generic approach to improve the properties of therapeutic proteins.
authors.affiliation: MedImmune Ltd, Granta Park, Cambridge, CB21 6GH, UK.
authors.forename: Claire L
authors.initials: CL
authors.lastname: Dobson
authors.affiliation: Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds, LS2 9JT, UK.|School of Molecular and Cellular Biology, University of Leeds, Leeds, LS2 9JT, UK.
authors.forename: Paul W A
authors.initials: PW
authors.lastname: Devine
authors.affiliation: Department of Chemical Engineering and Biotechnology, University of Cambridge, Cambridge, CB2 3RA, UK.
authors.forename: Jonathan J
authors.initials: JJ
authors.lastname: Phillips
authors.affiliation: MedImmune Ltd, Granta Park, Cambridge, CB21 6GH, UK.
authors.forename: Daniel R
authors.initials: DR
authors.lastname: Higazi
authors.affiliation: MedImmune Ltd, Granta Park, Cambridge, CB21 6GH, UK.
authors.forename: Christopher
authors.initials: C
authors.lastname: Lloyd
authors.affiliation: MedImmune Ltd, Granta Park, Cambridge, CB21 6GH, UK.
authors.forename: Bojana
authors.initials: B
authors.lastname: Popovic
authors.affiliation: MedImmune Ltd, Granta Park, Cambridge, CB21 6GH, UK.
authors.forename: Joanne
authors.initials: J
authors.lastname: Arnold
authors.affiliation: MedImmune Ltd, Granta Park, Cambridge, CB21 6GH, UK.
authors.forename: Andrew
authors.initials: A
authors.lastname: Buchanan
authors.affiliation: MedImmune Ltd, Granta Park, Cambridge, CB21 6GH, UK.
authors.forename: Arthur
authors.initials: A
authors.lastname: Lewis
authors.affiliation: MedImmune Ltd, Granta Park, Cambridge, CB21 6GH, UK.
authors.forename: Joanne
authors.initials: J
authors.lastname: Goodman
authors.affiliation: MedImmune Ltd, Granta Park, Cambridge, CB21 6GH, UK.
authors.forename: Christopher F
authors.initials: CF
authors.lastname: van der Walle
authors.affiliation: MedImmune Ltd, Granta Park, Cambridge, CB21 6GH, UK.
authors.forename: Peter
authors.initials: P
authors.lastname: Thornton
authors.affiliation: MedImmune Ltd, Granta Park, Cambridge, CB21 6GH, UK.
authors.forename: Lisa
authors.initials: L
authors.lastname: Vinall
authors.affiliation: MedImmune Ltd, Granta Park, Cambridge, CB21 6GH, UK.
authors.forename: David
authors.initials: D
authors.lastname: Lowne
authors.affiliation: Discovery Sciences, Innovative Medicines and Early Development, AstraZeneca, Pepparedsleden 1, Mölndal, 43183, Sweden.
authors.forename: Anna
authors.initials: A
authors.lastname: Aagaard
authors.affiliation: Discovery Sciences, Innovative Medicines and Early Development, AstraZeneca, Pepparedsleden 1, Mölndal, 43183, Sweden.
authors.forename: Lise-Lotte
authors.initials: LL
authors.lastname: Olsson
authors.affiliation: Discovery Sciences, Innovative Medicines and Early Development, AstraZeneca, Pepparedsleden 1, Mölndal, 43183, Sweden.
authors.forename: Anna
authors.initials: A
authors.lastname: Ridderstad Wollberg
authors.affiliation: MedImmune Ltd, Granta Park, Cambridge, CB21 6GH, UK.
authors.forename: Fraser
authors.initials: F
authors.lastname: Welsh
authors.affiliation: Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds, LS2 9JT, UK.|School of Molecular and Cellular Biology, University of Leeds, Leeds, LS2 9JT, UK.
authors.forename: Theodoros K
authors.initials: TK
authors.lastname: Karamanos
authors.affiliation: Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds, LS2 9JT, UK.|School of Molecular and Cellular Biology, University of Leeds, Leeds, LS2 9JT, UK.
authors.forename: Clare L
authors.initials: CL
authors.lastname: Pashley
authors.affiliation: Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds, LS2 9JT, UK.|School of Molecular and Cellular Biology, University of Leeds, Leeds, LS2 9JT, UK.
authors.forename: Matthew G
authors.initials: MG
authors.lastname: Iadanza
authors.affiliation: Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds, LS2 9JT, UK.|School of Molecular and Cellular Biology, University of Leeds, Leeds, LS2 9JT, UK.
authors.forename: Neil A
authors.initials: NA
authors.lastname: Ranson
authors.affiliation: Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds, LS2 9JT, UK.|School of Molecular and Cellular Biology, University of Leeds, Leeds, LS2 9JT, UK.
authors.forename: Alison E
authors.initials: AE
authors.lastname: Ashcroft
authors.affiliation: MedImmune Ltd, Granta Park, Cambridge, CB21 6GH, UK.
authors.forename: Alistair D
authors.initials: AD
authors.lastname: Kippen
authors.affiliation: MedImmune Ltd, Granta Park, Cambridge, CB21 6GH, UK.
authors.forename: Tristan J
authors.initials: TJ
authors.lastname: Vaughan
authors.affiliation: Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds, LS2 9JT, UK.|School of Molecular and Cellular Biology, University of Leeds, Leeds, LS2 9JT, UK.
authors.forename: Sheena E
authors.initials: SE
authors.lastname: Radford
authors.affiliation: MedImmune Ltd, Granta Park, Cambridge, CB21 6GH, UK.
authors.forename: David C
authors.initials: DC
authors.lastname: Lowe
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000378:metabolism* / Q000493:pharmacokinetics; D055592:Biophysical Phenomena; D002850:Chromatography, Gel; D004797:Enzyme-Linked Immunosorbent Assay; D057809:HEK293 Cells; D006801:Humans; D006859:Hydrogen; D051379:Mice; D009154:Mutation / Q000235:genetics; D009928:Organ Specificity; D011485:Protein Binding; D011487:Protein Conformation; D015202:Protein Engineering / Q000379:methods*; D025941:Protein Interaction Mapping; D055503:Protein Multimerization; D051381:Rats; D021241:Spectrometry, Mass, Electrospray Ionization; D013499:Surface Properties; D014783:Viscosity
chemical_list: D000911:Antibodies, Monoclonal; D006859:Hydrogen