Implementation of a LC-MS based multi-attribute method (MAM) and intact multi-attribute method (iMAM) workflow for...

Area of Science:

• Analytical Chemistry
• Biopharmaceutical Analysis
• Mass Spectrometry

Background:

• Mass spectrometry (MS) is increasingly adopted in Quality Control (QC) laboratories.
• The multi-attribute method (MAM) and intact multi-attribute method (iMAM) are advanced liquid chromatography-mass spectrometry (LC-MS) techniques.
• These methods aim to consolidate multiple conventional assays into a single LC-MS workflow for biotherapeutic protein analysis.

Purpose of the Study:

• To evaluate the application of MAM and iMAM for analyzing a glucagon-like peptide-Fc fusion protein.
• To demonstrate the capability of these methods in identifying and monitoring critical quality attributes (CQAs).
• To assess the suitability of MAM and iMAM workflows within current Good Manufacturing Practices (cGMP) environments.

Main Methods:

• Utilized both MAM and iMAM workflows for the analysis of four batches of a glucagon-like peptide-Fc fusion protein.
• MAM involved a discovery phase for CQA identification and a target monitoring phase.
• Optimized size exclusion and strong cation exchange chromatography for the native iMAM workflow to monitor intact-level CQAs.

Main Results:

• Successfully identified and monitored critical quality attributes (CQAs) for the biotherapeutic protein using both MAM and iMAM.
• New peak detection analysis identified changes in the protein attributes across the analyzed batches.
• Demonstrated the ability to monitor CQAs at both the peptide and intact protein levels.

Conclusions:

• MAM and iMAM are effective and compliant methods for comprehensive biotherapeutic protein characterization in QC settings.
• These LC-MS based methods can replace or supplement traditional assays, enhancing efficiency and data quality.
• The implementation of MAM and iMAM is feasible within CFR 11 compliant environments, supporting regulatory requirements.