An evaluation of instrument types for mass spectrometry-based multi-attribute analysis of biotherapeutics

Abstract

Multi-attribute methods (MAM), based on proteolytic digestion followed by liquid chromatography-mass spectrometry analysis of proteolytic peptides, have gained substantial attention in the biopharmaceutical industry for quantifying a variety of quality attributes for therapeutic proteins. Most MAM developed so far have been based on high-resolution mass spectrometers, due to their superb resolving power to distinguish analyte signals from interferences. Lower-resolution instruments, if demonstrated suitable, may further promote the adoption of the technology due to their low cost, small footprint, and ease of use. In this work, we compared the performance of a high-resolution instrument with a few low-resolution quadrupole-type instruments in quantifying a diverse set of quality attributes in a monoclonal antibody product. Different modes of operation for the quadrupole instruments, including scan mode, selected-ion monitoring and multiple-reaction monitoring, were evaluated. The high-resolution instrument has superb performance, with a quantitation limit of 0.002%. Single-quadrupole instruments in scan mode, on the other hand, provide a quantitation limit of about 1%, which may be fit-for-purpose for many routine MAM applications.

Interlaboratory Studies Using the NISTmAb to Advance Biopharmaceutical Structural Analytics

“…The multi-attribute method (MAM) builds upon industry experience with mass spectrometry (MS)-based peptide mapping ( Formolo et al, 2015 ; Rogstad et al, 2017 ; Noor et al, 2020 ) and holds promise for use in the quality control (QC) space ( Rogers et al, 2013 ; Rogers et al, 2015 ; Xu et al, 2017 ; Zhang and Guo, 2017 ; Rogstad et al, 2019 ; Millán-Martín et al, 2020 ; Sokolowska et al, 2020 ; Zhang et al, 2020 ; Mouchahoir et al, 2021 ). MAM is designed to monitor the status of pre-defined quality attributes within a therapeutic protein sample ( e.g ., post-translational modifications (PTMs), enzymatic clips, isomerization, etc. )…”

New Peak Detection Performance Metrics from the MAM Consortium Interlaboratory Study

“…Although MS-based platforms have traditionally been confined to the process development lab, the concept of utilizing MS in the QC lab through the application of MAM as a release method was introduced in 2013 . Indeed, a number of recent reports indicate a continuing interest in the implementation of mass spectrometry in the quality lab, ,− and the transfer of a robust and accurate peptide-mapping method between four locations has been demonstrated as a preliminary step toward implementing MAM in the QC environment …”

Multi attribute method implementation using a High Resolution Mass Spectrometry platform: From sample preparation to batch analysis

“…The workflow of this bottom-up methodology consists of an enzymatic digestion, followed by peptide separation and MS detection. Within peptide mapping methods, multi-attribute method (MAM) approaches are becoming an emerging tool [ 6 – 8 , 18 ]. MAM liquid chromatography (LC)-MS based method was firstly developed by Rogers et al [ 9 ] with the purpose to monitor simultaneously several critical attributes (chemical modifications) using only one method.…”

“…MAM liquid chromatography (LC)-MS based method was firstly developed by Rogers et al [ 9 ] with the purpose to monitor simultaneously several critical attributes (chemical modifications) using only one method. Due to its promising potential, it has gained popularity and interest and several pharmaceutical companies have also developed and implemented MAM workflows [ 8 , 18 ].…”

“…Therefore, by streamlining monitoring and quality control analysis, MAM has the potential to accelerate mAbs’ process development and manufacturing, contributing to reduce the risk of failure and costs associated with analytical characterization during bioprocess development, product release, and in-process control [ 6 , 9 , 22 ]. MAM has been increasingly used for routine PQA analysis [ 18 ] keeping up with the growth and increased acceptance of MS methods in the biopharmaceutical field. In fact, according to a recent US Food and Drug Administration study, most of the biotherapeutics (with license applications approved between 2000 and 2015) use MS methods for their characterization.…”