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Development of an efficient mAb quantification assay by LC-MS/MS using rapid on-bead digestion
Huai-Hsuan Chiu1, Yun-Jung Tsai1, Chiao Lo2
1School of Pharmacy, College of Medicine, National Taiwan University, Taipei, Taiwan; The Metabolomics Core Laboratory, Centers of Genomic and Precision Medicine, National Taiwan University, Taipei, Taiwan.
Analytica Chimica Acta
|January 21, 2022
Summary
A new, efficient liquid chromatography-tandem mass spectrometry method simplifies quantifying monoclonal antibody (mAb) levels in patients. This technique supports personalized medicine by enabling faster therapeutic drug monitoring for better treatment responses.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Pharmacology
Background:
- Monoclonal antibody (mAb) therapeutics are increasingly used, but individual concentration variations impact treatment efficacy.
- Personalized dosing for mAb therapies is crucial but challenging due to current quantification method limitations.
- Existing methods for quantifying mAbs using LC-MS/MS are often time-consuming and require extensive sample preparation.
Purpose of the Study:
- To develop a more efficient and practical LC-MS/MS method for quantifying multiple mAb therapeutics.
- To optimize an on-bead trypsin digestion protocol for faster and more reproducible mAb analysis.
- To validate the developed method for potential use in routine therapeutic drug monitoring and personalized medicine.
Main Methods:
- Developed an optimized on-bead trypsin digestion protocol at an elevated temperature (60°C for 60 min).
- Quantified five different mAbs (bevacizumab, evolocumab, nivolumab, pembrolizumab, trastuzumab) using LC-MS/MS with tocilizumab as the internal standard.
- Compared the on-bead digestion method with conventional low-pH elution, assessing sensitivity and reproducibility.
Main Results:
- The optimized on-bead digestion method demonstrated improved sensitivity and reproducibility compared to the conventional method for most mAbs.
- Achieved high accuracy (within 94.5% to 111.6%) and precision (RSD < 6.1% for repeatability, < 9.5% for intermediate precision) for the quantified mAbs.
- Successfully applied the method to measure trastuzumab concentrations in breast cancer patients, ranging from 66.4 to 173.2 μg/mL.
Conclusions:
- The developed LC-MS/MS method with on-bead digestion is efficient and practical for analyzing numerous clinical samples.
- This method facilitates routine therapeutic drug monitoring, contributing to personalized mAb treatment strategies.
- The improved quantification approach supports precision medicine by enabling better management of mAb drug exposure and treatment response.
Keywords:
High temperature trypsin digestionLiquid chromatography–tandem mass spectrometry (LC-MS/MS)Monoclonal antibody (mAb)On-bead trypsin digestionProtein G purificationTherapeutic drug monitoring (TDM)
