title: Monoclonal antibody interactions with micro- and nanoparticles: adsorption, aggregation, and accelerated stress studies.
abstract: Therapeutic proteins are exposed to various wetted surfaces that could shed subvisible particles. In this work we measured the adsorption of a monoclonal antibody (mAb) to various microparticles, characterized the adsorbed mAb secondary structure, and determined the reversibility of adsorption. We also developed and used a front-face fluorescence quenching method to determine that the mAb tertiary structure was near-native when adsorbed to glass, cellulose, and silica. Initial adsorption to each of the materials tested was rapid. During incubation studies, exposure to the air-water interface was a significant cause of aggregation but acted independently of the effects of microparticles. Incubations with glass, cellulose, stainless steel, or Fe(2)O(3) microparticles gave very different results. Cellulose preferentially adsorbed aggregates from solution. Glass and Fe(2)O(3) adsorbed the mAb but did not cause aggregation. Adsorption to stainless steel microparticles was irreversible, and caused appearance of soluble aggregates upon incubation. The secondary structure of mAb adsorbed to glass and cellulose was near-native. We suggest that the protocol described in this work could be a useful preformulation stress screening tool to determine the sensitivity of a therapeutic protein to exposure to common surfaces encountered during processing and storage.
authors.affiliation: Department of Chemical and Biological Engineering, University of Colorado, Boulder, Colorado 80309, USA.
authors.forename: Jared S
authors.initials: JS
authors.lastname: Bee
authors.forename: David
authors.initials: D
authors.lastname: Chiu
authors.forename: Suzanne
authors.initials: S
authors.lastname: Sawicki
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Stevenson
authors.forename: Koustuv
authors.initials: K
authors.lastname: Chatterjee
authors.forename: Erwin
authors.initials: E
authors.lastname: Freund
authors.forename: John F
authors.initials: JF
authors.lastname: Carpenter
authors.forename: Theodore W
authors.initials: TW
authors.lastname: Randolph
mesh_terms: D000327:Adsorption; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry* / Q000276:immunology; D008863:Microspheres; D053758:Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D010316:Particle Size; D055503:Protein Multimerization; D055550:Protein Stability; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D012995:Solubility; D013055:Spectrophotometry, Infrared; D013232:Steel / Q000737:chemistry; D019809:Streptavidin / Q000276:immunology*; D014867:Water / Q000737:chemistry
chemical_list: D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D014867:Water; D013232:Steel; D019809:Streptavidin