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

Area of Science:

• Biopharmaceutical quality control
• Analytical chemistry
• Mass spectrometry

Background:

• Biopharmaceutical quality control is increasingly complex due to evolving regulatory demands and intricate product structures.
• Traditional methods for assessing product quality attributes (PQAs) are often time-consuming and costly.
• Mass Spectrometry (MS)-based methods, particularly the multi-attribute method (MAM), are emerging as powerful tools for comprehensive biopharmaceutical characterization.

Purpose of the Study:

• To describe the implementation process of a Multi-Attribute Method (MAM) using a QTOF high-resolution mass spectrometry platform.
• To demonstrate the capability of MAM for simultaneous monitoring of various Product Quality Attributes (PQAs) in monoclonal antibodies (mAbs).
• To discuss troubleshooting for sample preparation and LC-MS parameters for high-throughput batch analysis.

Main Methods:

• Implementation of a MAM workflow on a QTOF high-resolution mass spectrometry platform.
• Utilized NIST mAb reference material and an in-process mAb sample for method validation.
• Employed high mass accuracy/high-resolution MS data for direct and simultaneous monitoring of PQAs.

Main Results:

• Successfully monitored critical PQAs including glycosylation profiles, methionine oxidation, tryptophan dioxidation, asparagine deamidation, N-terminal pyro-glutamic acid formation, and glycation.
• The MAM workflow proved effective for comprehensive characterization of a downstream processing polishing platform.
• Demonstrated the utility of MAM in a comparability study following technology transfer between laboratories.

Conclusions:

• The described MAM implementation provides a robust and efficient workflow for biopharmaceutical quality control.
• MAM offers a streamlined approach, replacing multiple orthogonal methods and reducing analysis time and costs.
• This high-resolution MS-based MAM serves as a valuable reference analytical tool for biopharmaceutical characterization and comparability studies.