title: Observation of small cluster formation in concentrated monoclonal antibody solutions and its implications to solution viscosity.
abstract: Monoclonal antibodies (mAbs) are a major class of biopharmaceuticals. It is hypothesized that some concentrated mAb solutions exhibit formation of a solution phase consisting of reversibly self-associated aggregates (or reversible clusters), which is speculated to be responsible for their distinct solution properties. Here, we report direct observation of reversible clusters in concentrated solutions of mAbs using neutron spin echo. Specifically, a stable mAb solution is studied across a transition from dispersed monomers in dilute solution to clustered states at more concentrated conditions, where clusters of a preferred size are observed. Once mAb clusters have formed, their size, in contrast to that observed in typical globular protein solutions, is observed to remain nearly constant over a wide range of concentrations. Our results not only conclusively establish a clear relationship between the undesirable high viscosity of some mAb solutions and the formation of reversible clusters with extended open structures, but also directly observe self-assembled mAb protein clusters of preferred small finite size similar to that in micelle formation that dominate the properties of concentrated mAb solutions.
authors.affiliation: Center for Neutron Research, National Institute of Standards and Technology, Gaithersburg, Maryland; Department of Chemical and Biomolecular Engineering, Center for Neutron Science, University of Delaware, Newark, Delaware.
authors.forename: Eric J
authors.initials: EJ
authors.lastname: Yearley
authors.affiliation: Center for Neutron Research, National Institute of Standards and Technology, Gaithersburg, Maryland; Department of Chemical and Biomolecular Engineering, Center for Neutron Science, University of Delaware, Newark, Delaware.
authors.forename: Paul D
authors.initials: PD
authors.lastname: Godfrin
authors.affiliation: Center for Neutron Research, National Institute of Standards and Technology, Gaithersburg, Maryland; Department of Chemical and Biomolecular Engineering, Center for Neutron Science, University of Delaware, Newark, Delaware.
authors.forename: Tatiana
authors.initials: T
authors.lastname: Perevozchikova
authors.affiliation: Center for Neutron Research, National Institute of Standards and Technology, Gaithersburg, Maryland; Institute for Research and Applied Physics, University of Maryland, College Park, Maryland.
authors.forename: Hailiang
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Institut Laue-Langevin, Grenoble, France.
authors.forename: Peter
authors.initials: P
authors.lastname: Falus
authors.affiliation: Institut Laue-Langevin, Grenoble, France.
authors.forename: Lionel
authors.initials: L
authors.lastname: Porcar
authors.affiliation: Center for Neutron Research, National Institute of Standards and Technology, Gaithersburg, Maryland; Center for Exploration of Energy and Matter, Indiana University, Bloomington, Indiana.
authors.forename: Michihiro
authors.initials: M
authors.lastname: Nagao
authors.affiliation: Center for Neutron Research, National Institute of Standards and Technology, Gaithersburg, Maryland.
authors.forename: Joseph E
authors.initials: JE
authors.lastname: Curtis
authors.affiliation: Theranos Inc., Palo Alto, California.
authors.forename: Pradad
authors.initials: P
authors.lastname: Gawande
authors.affiliation: Late Stage Pharmaceutical Development, Genentech Inc., South San Francisco, California.
authors.forename: Rosalynn
authors.initials: R
authors.lastname: Taing
authors.affiliation: Late Stage Pharmaceutical Development, Genentech Inc., South San Francisco, California. Electronic address: zarraga.isidro@gene.com.
authors.forename: Isidro E
authors.initials: IE
authors.lastname: Zarraga
authors.affiliation: Department of Chemical and Biomolecular Engineering, Center for Neutron Science, University of Delaware, Newark, Delaware.
authors.forename: Norman J
authors.initials: NJ
authors.lastname: Wagner
authors.affiliation: Center for Neutron Research, National Institute of Standards and Technology, Gaithersburg, Maryland; Department of Chemical and Biomolecular Engineering, Center for Neutron Science, University of Delaware, Newark, Delaware. Electronic address: yunliu@udel.edu.
authors.forename: Yun
authors.initials: Y
authors.lastname: Liu
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry*; D004573:Electrolytes / Q000737:chemistry; D006801:Humans; D008958:Models, Molecular; D066329:Protein Aggregates; D055503:Protein Multimerization; D012492:Salts / Q000737:chemistry; D053838:Scattering, Small Angle; D012996:Solutions; D014783:Viscosity; D014961:X-Ray Diffraction
chemical_list: D000911:Antibodies, Monoclonal; D004573:Electrolytes; D066329:Protein Aggregates; D012492:Salts; D012996:Solutions
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