title: Competing aggregation pathways for monoclonal antibodies.
abstract: Aggregation is mediated by local unfolding to allow aggregation "hot spot(s)" to become solvent exposed and available to associate with a hot spot on another partially unfolded protein. Historically, the unfolding of either the crystallizable fragment (Fc) or the antigen binding fragment (Fab) regions of a given monoclonal antibody (MAb) has been implicated in aggregation, with differing results across different proteins. The present work focuses on separately quantifying the aggregation kinetics of isolated Fc, isolated Fab, and intact MAb as a function of pH under accelerated (high temperature) conditions. The results show that both Fab and Fc are aggregation prone and compete within the same MAb.
authors: Department of Chemistry and Biochemistry, University of Delaware, Newark, DE 19711, USA; Department of Biotherapeutics, Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, CT 06877, USA.
authors: Haixia
authors: H
authors: Wu
authors: Department of Biotherapeutics, Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, CT 06877, USA.
authors: Rachel
authors: R
authors: Kroe-Barrett
authors: Department of Biotherapeutics, Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, CT 06877, USA.
authors: Sanjaya
authors: S
authors: Singh
authors: Department of Chemical and Biomolecular Engineering, Tulane University, New Orleans, LA 70118, USA; Department of Chemical and Biomolecular Engineering, University of Delaware, Newark, DE 19711, USA.
authors: Anne S
authors: AS
authors: Robinson
authors: Department of Chemical and Biomolecular Engineering, University of Delaware, Newark, DE 19711, USA. Electronic address: cjr@udel.edu.
authors: Christopher J
authors: CJ
authors: Roberts
mesh_terms: D000818:Animals; D058846:Antibodies, Monoclonal, Murine-Derived / Q000737:chemistry*; D019013:CD40 Antigens / Q000276:immunology; D002152:Calorimetry, Differential Scanning; D002850:Chromatography, Gel; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry / Q000302:isolation & purification; D007141:Immunoglobulin Fc Fragments / Q000737:chemistry / Q000302:isolation & purification; D007074:Immunoglobulin G / Q000737:chemistry*; D007700:Kinetics; D051379:Mice; D017510:Protein Folding; D055503:Protein Multimerization; D059748:Proteolysis
chemical_list: D058846:Antibodies, Monoclonal, Murine-Derived; D019013:CD40 Antigens; D007140:Immunoglobulin Fab Fragments; D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G
keywords: Aggregation; Hot-spot; MAb