Development, validation, and implementation of capillary gel electrophoresis as a replacement for SDS-PAGE for purity analysis of IgG2 mAbs.
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.
Lacher
Nathan A
NA
Analytical R&D, Pfizer BioTherapeutics R&D, Chesterfield, MO 63017, USA.
Roberts
Rachel K
RK
He
Yan
Y
Cargill
Holly
H
Kearns
Katrina M
KM
Holovics
Heidi
H
Ruesch
Margaret N
MN
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
D000477:Alkylating Agents; D000911:Antibodies, Monoclonal; D002021:Buffers; D007074:Immunoglobulin G; D019163:Reducing Agents; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase