title: Extensive Chemical Modifications in the Primary Protein Structure of IgG1 Subvisible Particles Are Necessary for Breaking Immune Tolerance.
abstract: A current concern with the use of therapeutic proteins is the likely presence of aggregates and submicrometer, subvisible, and visible particles. It has been proposed that aggregates and particles may lead to unwanted increases in the immune response with a possible impact on safety or efficacy. The aim of this study was thus to evaluate the ability of subvisible particles of a therapeutic antibody to break immune tolerance in an IgG1 transgenic mouse model and to understand the particle attributes that might play a role in this process. We investigated the immunogenic properties of subvisible particles (unfractionated, mixed populations, and well-defined particle size fractions) using a transgenic mouse model expressing a mini-repertoire of human IgG1 (hIgG1 tg). Immunization with proteinaceous subvisible particles generated by artificial stress conditions demonstrated that only subvisible particles bearing very extensive chemical modifications within the primary amino acid structure could break immune tolerance in the hIgG1 transgenic mouse model. Protein particles exhibiting low levels of chemical modification were not immunogenic in this model.
authors.affiliation: Analytical Development & Quality Control, Pharma Technical Development Biologics Europe, F. Hoffmann-La Roche Ltd. , Basel, Switzerland.|Division of Pharmaceutical Technology, Department of Pharmaceutical Sciences, University of Basel , Basel, Switzerland.
authors.forename: Björn
authors.initials: B
authors.lastname: Boll
authors.affiliation: Roche Pharmaceutical Research and Early Development , Pharmaceutical Sciences, Roche Innovation Center, Basel, Switzerland.
authors.forename: Juliana
authors.initials: J
authors.lastname: Bessa
authors.affiliation: Analytical Development & Quality Control, Pharma Technical Development Biologics Europe, F. Hoffmann-La Roche Ltd. , Basel, Switzerland.|Pharmaceutical Development & Supplies, Pharma Technical Development Biologics Europe, F. Hoffmann-La Roche Ltd. , Basel, Switzerland.|Division of Pharmaceutical Technology, Department of Pharmaceutical Sciences, University of Basel , Basel, Switzerland.
authors.forename: Emilien
authors.initials: E
authors.lastname: Folzer
authors.affiliation: Analytical Development & Quality Control, Pharma Technical Development Biologics Europe, F. Hoffmann-La Roche Ltd. , Basel, Switzerland.|Division of Pharmaceutical Technology, Department of Pharmaceutical Sciences, University of Basel , Basel, Switzerland.
authors.forename: Anacelia
authors.identifier: 0000-0002-9577-4839
authors.initials: A
authors.lastname: Ríos Quiroz
authors.affiliation: Pharmaceutical Development & Supplies, Pharma Technical Development Biologics Europe, F. Hoffmann-La Roche Ltd. , Basel, Switzerland.
authors.forename: Roland
authors.initials: R
authors.lastname: Schmidt
authors.affiliation: Analytical Development & Quality Control, Pharma Technical Development Biologics Europe, Roche Diagnostics GmbH , Penzberg, Germany.
authors.forename: Patrick
authors.initials: P
authors.lastname: Bulau
authors.affiliation: Analytical Development & Quality Control, Pharma Technical Development Biologics Europe, F. Hoffmann-La Roche Ltd. , Basel, Switzerland.
authors.forename: Christof
authors.initials: C
authors.lastname: Finkler
authors.affiliation: Pharmaceutical Development & Supplies, Pharma Technical Development Biologics Europe, F. Hoffmann-La Roche Ltd. , Basel, Switzerland.
authors.forename: Hanns-Christian
authors.initials: HC
authors.lastname: Mahler
authors.affiliation: Division of Pharmaceutical Technology, Department of Pharmaceutical Sciences, University of Basel , Basel, Switzerland.
authors.forename: Jörg
authors.initials: J
authors.lastname: Huwyler
authors.affiliation: Roche Pharmaceutical Research and Early Development , Pharmaceutical Sciences, Roche Innovation Center, Basel, Switzerland.
authors.forename: Antonio
authors.initials: A
authors.lastname: Iglesias
authors.affiliation: Analytical Development & Quality Control, Pharma Technical Development Biologics Europe, F. Hoffmann-La Roche Ltd. , Basel, Switzerland.
authors.forename: Atanas V
authors.initials: AV
authors.lastname: Koulov
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D000917:Antibody Formation / Q000276:immunology; D006801:Humans; D007108:Immune Tolerance / Q000276:immunology*; D007074:Immunoglobulin G / Q000737:chemistry*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D010316:Particle Size
chemical_list: D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G
keywords: IgG1 tg mouse; biotherapeutics; chemical modifications; immunogenicity; subvisible protein particles