LC/MS Methods for Studying Lysosomal ADC Catabolism.

Area of Science:

• Biochemistry
• Pharmacology
• Drug Development

Background:

• Antibody-drug conjugates (ADCs) require characterization of the active released drug entity.
• Cellular uptake and lysosomal enzyme activity are key factors in ADC efficacy.
• Understanding payload release is crucial for identifying potential liabilities.

Purpose of the Study:

• To describe a general in vitro method for analyzing the release characteristics of ADCs.
• To utilize commercial human lysosomal preparations coupled with LC/MS for this analysis.
• To identify active released entities and assess payload stability in ADCs.

Main Methods:

• Employing commercial human lysosomal preparations as an in vitro tool.
• Utilizing liquid chromatography-mass spectrometry (LC/MS) for analysis.
• Developing a general method applicable to both cathepsin B-cleavable and non-cleavable ADCs.

Main Results:

• Demonstrated the utility of human lysosomal preparations for exploring ADC release.
• Provided insights into the release characteristics of various ADCs.
• Identified potential metabolic or chemical liabilities of new payload structures.

Conclusions:

• The described in vitro method is valuable for antibody-drug conjugate development.
• Human lysosomal preparations coupled with LC/MS offer a robust approach to study ADC payload release.
• The method allows for tailored investigations based on specific ADC designs and payload types.