Establishing Multi-Dimensional LC-MS Systems for Versatile Workflows to Analyze Therapeutic Antibodies at Different Molecular Levels in Routine Operations DOI Creative Commons
Katrin Heinrich, Sina Hoelterhoff, Saban Oezipek

et al.

Pharmaceuticals, Journal Year: 2025, Volume and Issue: 18(3), P. 401 - 401

Published: March 12, 2025

Background/Objectives: Multi-dimensional liquid chromatography coupled with mass spectrometry (mD-LC-MS) has emerged as a powerful technique for the in-depth characterization of biopharmaceuticals by assessing chromatographically resolved product variants in streamlined and semi-automated manner. The study aims to demystify enhance accessibility this but inherently complex detailing robust user-friendly instrument platform, allowing analysts switch seamlessly between intact, subunit, peptide mapping workflows. Methods: Starting from commercially available Two-Dimensional Liquid Chromatography (2D-LC) system, we introduce specific hardware software extensions leading two versatile mD-LC-MS setups, slightly different configurations. technique’s efficacy is demonstrated through case on cation exchange method charge bispecific antibody, isolating peak(s) interest, followed online sample processing, including reduction enzymatic digestion, subsequently analysis. Results: accuracy reproducibility both setups proposed were successfully tested. Despite peak patterns first dimension, systems equally effective identifying quantifying underlying species. This highlights routine usability technology (ultra) high-performance (UHPLC) therapeutic biomolecule. Conclusions: reliability underscore practicality use biopharmaceutical Our detailed description insights simplify access advanced broader scientific community, regardless expertise level, lower entry barrier its various research industrial settings.

Language: Английский

Establishing Multi-Dimensional LC-MS Systems for Versatile Workflows to Analyze Therapeutic Antibodies at Different Molecular Levels in Routine Operations DOI Creative Commons
Katrin Heinrich, Sina Hoelterhoff, Saban Oezipek

et al.

Pharmaceuticals, Journal Year: 2025, Volume and Issue: 18(3), P. 401 - 401

Published: March 12, 2025

Background/Objectives: Multi-dimensional liquid chromatography coupled with mass spectrometry (mD-LC-MS) has emerged as a powerful technique for the in-depth characterization of biopharmaceuticals by assessing chromatographically resolved product variants in streamlined and semi-automated manner. The study aims to demystify enhance accessibility this but inherently complex detailing robust user-friendly instrument platform, allowing analysts switch seamlessly between intact, subunit, peptide mapping workflows. Methods: Starting from commercially available Two-Dimensional Liquid Chromatography (2D-LC) system, we introduce specific hardware software extensions leading two versatile mD-LC-MS setups, slightly different configurations. technique’s efficacy is demonstrated through case on cation exchange method charge bispecific antibody, isolating peak(s) interest, followed online sample processing, including reduction enzymatic digestion, subsequently analysis. Results: accuracy reproducibility both setups proposed were successfully tested. Despite peak patterns first dimension, systems equally effective identifying quantifying underlying species. This highlights routine usability technology (ultra) high-performance (UHPLC) therapeutic biomolecule. Conclusions: reliability underscore practicality use biopharmaceutical Our detailed description insights simplify access advanced broader scientific community, regardless expertise level, lower entry barrier its various research industrial settings.

Language: Английский

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