Title:Interlaboratory Co-validation of a UPLC-ToF MS MAM Method for
Truncations of a Fc Fusion Protein
Volume: 24
Issue: 10
Author(s): Gang Wu, Chuanfei Yu, Qinghe Tong, Wenbo Wang, Xi Huang, Hao Li and Lan Wang*
Affiliation:
- Division of Monoclonal Antibody Product, National Institute for Food and Drug Control, Beijing, China
Keywords:
Interlaboratory, method validation, MAM, multi-attribute method, UPLC-ToF MS, fusion protein, truncation, LCMS, HRMS, subunit, CQAs, QC release testing, stability.
Abstract:
Background: Peptide-Fc fusion proteins are inherently heterogeneous and complex molecules.
Protein post-translational modifications (PTMs) or truncation can arise during manufacturing
or product storage. Some of these product attributes could potentially impact the efficacy or safety of
the bio-molecule and are thus classified as critical quality attributes (CQAs). These CQAs should be
controlled in order to ensure manufacturing and quality consistency.
Methods: A subunit UPLC-ToF MS based MAM method was developed for identity test and quantitatively
monitored two critical quality attributes (CQAs) resulting from two truncations of that fusion
protein (fragment 1 and 2). Three independent laboratories are involved in the method validation
according to ICH Q2(R1), ICH Q6B, FDA and NMPA guidance.
Results: This developed method fully meets the pre-defined analytical target profile (ATP), including
specificity, accuracy, precision, quantitation limit, linearity, range and robustness. Three independent
labs co-validate a UPLC-ToF MS based MAM method for protein drug QC release and stability
testing.
Conclusion: The experimental design of method validation can be a reference for LC-HRMS-based
subunit MAM methods that have been widely used in the characterization of antibodies, ADCs and
other protein-based biologics. This work paves the way for implementing MAM in QC with more
targeted control of product quality.
