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A simple and highly sensitive LC-MS workflow for characterization and quantification of ADC cleavable payloads
Shi Ya Mak1, Shuwen Chen1, Wey Jia Fong1
1Bioprocessing Technology Institute (BTI), Agency for Science, Technology and Research (A*STAR), Centros, Singapore, 138668, Singapore.
Scientific Reports
|May 14, 2024
Summary
A new, sensitive method quantifies six antibody-drug conjugate (ADC) payloads in mouse serum using liquid chromatography-tandem mass spectrometry. This validated workflow requires minimal sample volume and is effective for pharmacokinetic studies.
Area of Science:
- Pharmacology
- Analytical Chemistry
- Biochemistry
Background:
- Antibody-drug conjugates (ADCs) utilize cytotoxic payloads for targeted cancer therapy.
- Quantification of these payloads is crucial for pharmacokinetic and efficacy studies.
- Existing methods may lack sensitivity or require larger sample volumes.
Purpose of the Study:
- To develop and validate a sensitive, simultaneous quantification method for six common ADC payloads.
- To establish a workflow suitable for pharmacokinetic analysis in mouse serum.
Main Methods:
- A streamlined sample extraction using a methanol-ethanol mixture.
- Rapid liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis.
- Validation in mouse serum with assessment of linearity, recovery, and sensitivity.
Main Results:
- Simultaneous quantification of SN-38, MTX, DXd, MMAE, MMAF, and Calicheamicin (CM).
- Achieved linear response ranges from 0.04-1000 nM depending on the payload.
- High recovery (>85%) and requirement of only 5 µL serum volume.
- Successful application in a pharmacokinetic study for MMAE quantification.
Conclusions:
- A robust and sensitive LC-MS/MS method for simultaneous ADC payload quantification has been validated.
- The method's high sensitivity and low sample volume requirement make it ideal for preclinical pharmacokinetic studies.
- This validated workflow supports the development and evaluation of novel ADC therapeutics.
