High-sensitivity MAM to support early-stage activities during biotherapeutic development

Area of Science:

• Biopharmaceutical Analysis
• Analytical Chemistry
• Mass Spectrometry

Background:

• Developing sensitive analytical methods for early-stage biotherapeutic development is challenging due to limited product knowledge and sample availability.
• Robust methods are crucial for candidate screening, process development, formulation, and stability studies.

Purpose of the Study:

• To present a high-sensitivity multi-attribute method (MAM) using nano-flow LC-MS/MS for characterizing monoclonal antibody (mAb)-based molecules.
• To evaluate the method's performance for detecting and quantifying product quality attributes (PQAs) and impurities.

Main Methods:

• Utilized nano-flow liquid chromatography-tandem mass spectrometry (LC-MS/MS) for high-resolution analysis.
• Employed Orbitrap-based mass detection within a CFR Part 11 compliant data system.
• Evaluated run-to-run variation, sensitivity (LOD/LOQ), and new peak detection (NPD) capabilities.

Main Results:

• Demonstrated low run-to-run variation for key analytical parameters using NISTmAb as a reference.
• Achieved high sensitivity (LOD/LOQ: 0.6-7.0 ng) for detecting low-abundance PQAs like deamidation and oxidation.
• Successfully applied the nano-flow MAM for in-process monitoring of PQAs and impurities in a chimeric IgG1 mAb.

Conclusions:

• The nano-flow MAM is a sensitive and robust approach for biotherapeutic characterization.
• The method is suitable for early-stage bioprocess development, offering potential for quality control (QC) applications.
• NPD capability aids in detecting low-level impurities during in-process testing.