Improved Mass Accuracy and Precision for Multi-Attribute Methods Using a New Internally Calibrated High Resolution...

Area of Science:

• Biopharmaceutical Development
• Analytical Chemistry
• Mass Spectrometry

Background:

• Ensuring biotherapeutic safety and efficacy requires understanding product quality attributes (PQAs) and their impact on stability.
• The multi-attribute method (MAM), utilizing LC-MS peptide mapping, supports biotherapeutic development by monitoring PQAs.

Purpose of the Study:

• To evaluate a low-maintenance, MAM-ready mass detector for monitoring PQAs in proteolytically digested proteins.
• To assess the instrument's mass accuracy, precision, sensitivity, and long-term stability for MAM applications.

Main Methods:

• Evaluation of a MAM-ready mass detector with optimized source parameters for relative quantitation.
• Utilized a built-in one-point mass calibration and on-the-fly mass correction for high-resolution mass spectrometry.
• Assessed peptide monitoring over 54 days with sub-1 ppm mass accuracies and <3 ppm mass tolerances.

Main Results:

• The evaluated mass detector demonstrated high sensitivity, minimal in-source fragmentation, and robust relative quantitation of PQAs.
• Achieved sub-1 ppm mass accuracies and <3 ppm mass tolerances for peptide monitoring over extended periods (54 days).
• LC-MS MAM data from the instrument showed strong agreement with previous MAM systems and conventional assays.

Conclusions:

• The evaluated MAM-ready mass detector is suitable for reliable PQA monitoring in biopharmaceutical development.
• Its high accuracy, precision, and stability support robust and specific peptide monitoring for quality control.
• This instrument offers a low-maintenance, high-performance solution for advancing biotherapeutic characterization.