title: Molecular Insights into the Thermal Stability of mAbs with Variable-Temperature Ion-Mobility Mass Spectrometry.
abstract: The aggregation of protein-based therapeutics such as monoclonal antibodies (mAbs) can affect the efficacy of the treatment and can even induce effects that are adverse to the patient. Protein engineering is used to shift the mAb away from an aggregation-prone state by increasing the thermodynamic stability of the native fold, which might in turn alter conformational flexibility. We have probed the thermal stability of three types of intact IgG molecules and two Fc-hinge fragments by using variable-temperature ion-mobility mass spectrometry (VT-IM-MS). We observed changes in the conformations of isolated proteins as a function of temperature (300-550 K). The observed differences in thermal stability between IgG subclasses can be rationalized in terms of changes to higher-order structural organization mitigated by the hinge region. VT-IM-MS provides insights into mAbs structural thermodynamics and is presented as a promising tool for thermal-stability studies for proteins of therapeutic interest.
authors.affiliation: MIB and School of Chemistry, The University of Manchester, 131 Princess Street, Manchester, M1 7DN, UK.
authors.forename: Kamila J
authors.initials: KJ
authors.lastname: Pacholarz
authors.affiliation: UCB Pharma, 216 Bath Road, Slough, SL1 3WE, UK.
authors.forename: Shirley J
authors.initials: SJ
authors.lastname: Peters
authors.affiliation: UCB Pharma, 216 Bath Road, Slough, SL1 3WE, UK.
authors.forename: Rachel A
authors.initials: RA
authors.lastname: Garlish
authors.affiliation: UCB Pharma, 216 Bath Road, Slough, SL1 3WE, UK.
authors.forename: Alistair J
authors.initials: AJ
authors.lastname: Henry
authors.affiliation: UCB Pharma, 216 Bath Road, Slough, SL1 3WE, UK.
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Taylor
authors.affiliation: UCB Pharma, 216 Bath Road, Slough, SL1 3WE, UK.
authors.forename: David P
authors.initials: DP
authors.lastname: Humphreys
authors.affiliation: MIB and School of Chemistry, The University of Manchester, 131 Princess Street, Manchester, M1 7DN, UK. perdita.barran@manchester.ac.uk.
authors.forename: Perdita E
authors.initials: PE
authors.lastname: Barran
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry*; D007128:Immunoglobulin Fragments / Q000737:chemistry; D007074:Immunoglobulin G / Q000737:chemistry; D013058:Mass Spectrometry; D011487:Protein Conformation; D055550:Protein Stability; D013696:Temperature
chemical_list: D000911:Antibodies, Monoclonal; D007128:Immunoglobulin Fragments; D007074:Immunoglobulin G
keywords: ion mobility; mAb; mass spectrometry; protein therapeutics; thermal stability