title: Mass spectrometry for the biophysical characterization of therapeutic monoclonal antibodies.
abstract: Monoclonal antibodies (mAbs) are powerful therapeutics, and their characterization has drawn considerable attention and urgency. Unlike small-molecule drugs (150-600 Da) that have rigid structures, mAbs (∼150 kDa) are engineered proteins that undergo complicated folding and can exist in a number of low-energy structures, posing a challenge for traditional methods in structural biology. Mass spectrometry (MS)-based biophysical characterization approaches can provide structural information, bringing high sensitivity, fast turnaround, and small sample consumption. This review outlines various MS-based strategies for protein biophysical characterization and then reviews how these strategies provide structural information of mAbs at the protein level (intact or top-down approaches), peptide, and residue level (bottom-up approaches), affording information on higher order structure, aggregation, and the nature of antibody complexes.
authors.affiliation: Department of Chemistry, Washington University in St. Louis, St. Louis, MO 63130, USA.
authors.forename: Hao
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Department of Chemistry, Washington University in St. Louis, St. Louis, MO 63130, USA.
authors.forename: Weidong
authors.initials: W
authors.lastname: Cui
authors.affiliation: Department of Chemistry, Washington University in St. Louis, St. Louis, MO 63130, USA. Electronic address: mgross@wustl.edu.
authors.forename: Michael L
authors.initials: ML
authors.lastname: Gross
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000627:therapeutic use*; D055592:Biophysical Phenomena; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*
chemical_list: D000911:Antibodies, Monoclonal
keywords: FPOP; Hydrogen/deuterium exchange; Ion mobility; Mass spectrometry; Monoclonal antibody; Native ESI; Protein footprinting; Top-down and bottom-up