A Robust Purity Method for Biotherapeutics Using New Peak Detection in an LC-MS-Based Multi-Attribute Method.

Area of Science:

• Analytical Chemistry
• Biopharmaceutical Analysis
• Mass Spectrometry

Background:

• Liquid chromatography-mass spectrometry (LC-MS) based multi-attribute methods (MAM) are crucial for biopharmaceutical analysis.
• Conventional detection methods have limitations in identifying subtle product quality changes.
• New Peak Detection (NPD) offers sensitive, unbiased attribute monitoring but faces challenges with false positives.

Purpose of the Study:

• To enhance the reliability and implementation of New Peak Detection (NPD) within LC-MS-based multi-attribute methods (MAM).
• To address the limitations of NPD, specifically false positives and artifacts, hindering its broad adoption in the biopharmaceutical industry.
• To establish a robust system suitability control strategy for NPD in purity testing.

Main Methods:

• Development of a novel NPD system suitability control strategy.
• Implementation of false positive curation and a known peak list concept.
• Utilization of a pairwise analysis approach for sample-reference comparison.
• Introduction of sequence variant co-mixes for experimental performance assessment.

Main Results:

• The developed NPD strategies significantly reduce false positives and artifacts.
• NPD demonstrated superior performance in detecting unexpected changes compared to conventional methods.
• The pairwise analysis and control strategy enhance the accuracy and reduce subjectivity in purity testing.
• Experimental validation using sequence variant co-mixes confirmed NPD's enhanced performance.

Conclusions:

• NPD, enhanced with novel control strategies, provides a reliable and sensitive method for biopharmaceutical purity testing.
• This approach minimizes subjectivity and analyst intervention, reducing the risk of overlooking critical product quality changes.
• NPD represents a significant advancement in quality control, offering a more objective and comprehensive purity assessment.