Interlaboratory Co-validation of a UPLC-ToF MS MAM Method for Truncations of a Fc Fusion Protein

Abstract

aims: In this work, we describe the development and interlaboratory co-validation of a middle-down UPLC-ToF MS MAM (multi-attribute method) analytical procedure for a fusion protein drug candidate CQAs control. background: Liquid chromatography mass spectrometry (LC-MS) is a good choice for the quality control of some biological product fragments which cannot be effectively controlled by traditional methods,such as CE-SDS, RP-HPLC. objective: To establish a new MS based MAM method for identity test and quantitatively monitor two critical quality attributes resulting from two truncations of that fusion protein. method: A subunit UPLC-ToF MS based MAM method was developed for identity test and quantitatively monitor two critical quality attributes 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. result: Result shows that the developed method fully meet the pre-defined analytical target profile (ATP), including specificity, accuracy, precision, quantitation limit, linearity, range and robustness. conclusion: This is the first report describing the implementation of analytical method lifecycle management concept, and three independent labs co-validate a UPLC-ToF MS based MAM method for protein drug QC release and stability testing. The experimental design of method validation can be a reference for LC-HRMS based subunit MAM methods that have been being widely used in characterization of antibody, ADCs and other protein based biologics. This work paves the way for implementing MAM in QC with more targeted control of product quality. other: None