Development, qualification, and application of a highly efficient and robust new peak detection workflow for the...

Area of Science:

• Biopharmaceutical Analysis
• Analytical Chemistry
• Quality Control

Background:

• Multi-attribute method (MAM) using liquid chromatography-mass spectrometry peptide mapping offers a promising alternative to traditional purity/impurity assays for protein biopharmaceuticals.
• Reliable detection of new, absent, or changed peptide species is crucial for ensuring drug quality, safety, and efficacy, necessitating advanced New Peak Detection (NPD) capabilities.

Purpose of the Study:

• To develop, qualify, and apply a highly efficient and robust NPD workflow for MAM peptide mapping.
• To validate the NPD workflow according to ICH Q2 guidelines for reliable impurity detection in biopharmaceuticals.

Main Methods:

• Development and implementation of a New Peak Detection (NPD) workflow within Genedata Expressionist® software.
• Rational design of detection thresholds and validation of the NPD workflow following ICH Q2 guidelines.
• Application of the workflow to stability testing of drug products and detection of unknown impurities in drug substances.

Main Results:

• The developed NPD workflow reliably detects relevant peptide species with relative abundance below 1%.
• The workflow demonstrated high efficiency and robustness, successfully identifying critical peptide changes without reporting false positive peaks.
• Case studies confirmed the workflow's capability in stability testing and identifying unknown impurities in drug substances.

Conclusions:

• The validated NPD workflow represents a significant advancement in utilizing MAM for quality control of protein biopharmaceuticals.
• This method enables sensitive and reliable identification of true positive peaks, improving the assessment of drug quality, safety, and efficacy.