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Peptide Identification Using Tandem Mass Spectrometry01:33

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Related Experiment Video

Updated: Jan 15, 2026

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

Thomas Pohl1, Patrick Sascha Merkle1, Sonja Hudelmaier1

  • 1Biologics Analytical Development, Technical Research & Development, Novartis Pharma AG, Basel, Switzerland.

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|October 14, 2025
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Summary

This study introduces a new peak detection (NPD) workflow for multi-attribute method (MAM) peptide mapping. This robust method reliably identifies critical peptide changes in biopharmaceuticals, enhancing drug quality control.

Keywords:
Analytical method validationbiopharmaceuticalmass spectrometrymulti-attribute methodnew peak detectionquality control

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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.