Generic method approaches for monoclonal antibody therapeutics analysis using both ligand binding and LC-MS/MS...

Area of Science:

• Biopharmaceutical analysis
• Analytical chemistry
• Mass spectrometry

Background:

• Monoclonal antibodies (mAbs) are crucial biotherapeutics for disease intervention.
• Conventional ligand-binding assays have limitations in mAb characterization.
• Liquid chromatography-tandem mass spectrometry (LC-MS/MS) offers an alternative analytical approach.

Purpose of the Study:

• To present a generic LC-MS/MS method applicable to multiple mAb candidates.
• To highlight the efficiency gains from a standardized analytical workflow.
• To discuss the adaptability of the method for different analytical needs.

Main Methods:

• Development of a common workflow for LC-MS/MS analysis.
• Selection of a common surrogate peptide for various IgG isotypes (IgG1, IgG2, IgG4).
• Utilization of a stable isotope-labeled internal standard for quantification.

Main Results:

• Demonstration of a translatable generic LC-MS/MS method.
• The method is adaptable from single-analyte to multiple-analyte analysis.
• The method can be transitioned from generic to specific applications.

Conclusions:

• A generic LC-MS/MS method significantly reduces analytical development time for mAb biotherapeutics.
• The proposed method offers flexibility and can be tailored for specific or multiple analyte quantification.
• This approach is particularly effective for common IgG isotypes.