title: Development, validation, and implementation of capillary gel electrophoresis as a replacement for SDS-PAGE for purity analysis of IgG2 mAbs.
abstract: Capillary gel electrophoresis (CGE) methods with UV detection were developed for reduced and non-reduced mAb analysis. These methods can be used to evaluate mAb purity, offering more reproducible quantitation compared with that of traditional SDS-PAGE methods. These CGE methods have been utilized as platform technology for bioprocess development, formulation development, mAb characterization, drug substance/drug product release testing as well as a required methodology for stability testing. We have found these CGE methods to be applicable across a platform of mAbs in preclinical and clinical development, with the majority of mAbs requiring no modification to the method conditions. This methodology has been ICH validated and transferred to several supporting organizations. The data presented herein describes the development of CGE methodology, platform application to mAb purity analysis, ICH validation, reliability metrics, and considerations on technology enhancement for improved performance and throughput.
authors.affiliation: Analytical R&D, Pfizer BioTherapeutics R&D, Chesterfield, MO 63017, USA.
authors.forename: Nathan A
authors.initials: NA
authors.lastname: Lacher
authors.forename: Rachel K
authors.initials: RK
authors.lastname: Roberts
authors.forename: Yan
authors.initials: Y
authors.lastname: He
authors.forename: Holly
authors.initials: H
authors.lastname: Cargill
authors.forename: Katrina M
authors.initials: KM
authors.lastname: Kearns
authors.forename: Heidi
authors.initials: H
authors.lastname: Holovics
authors.forename: Margaret N
authors.initials: MN
authors.lastname: Ruesch
mesh_terms: D000477:Alkylating Agents / Q000737:chemistry; D000911:Antibodies, Monoclonal / Q000032:analysis* / Q000737:chemistry; D002021:Buffers; D053801:Capillary Electrochromatography / Q000379:methods*; D004355:Drug Stability; D004591:Electrophoresis, Polyacrylamide Gel; D019544:Equipment Failure Analysis; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000032:analysis* / Q000737:chemistry; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / Q000737:chemistry; D019163:Reducing Agents / Q000737:chemistry; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet; D013696:Temperature
chemical_list: D000477:Alkylating Agents; D000911:Antibodies, Monoclonal; D002021:Buffers; D007074:Immunoglobulin G; D019163:Reducing Agents; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase