Rapid and Simultaneous Characterization of Drug Conjugation in Heavy and Light Chains of a Monoclonal Antibody Revealed by High-Resolution Ion Mobility Separations in SLIM.

Analytical Chemistry
Gabe NagyYehia M Ibrahim

Abstract

Antibody-drug conjugates (ADCs) have recently gained traction in the biomedical community due to their promise for human therapeutics and an alternative to chemotherapy for cancer. Crucial metrics for ADC efficacy, safety, and selectivity are their drug-antibody ratios (DARs). However, DAR characterization (i.e., determining the average number of conjugated drugs on the antibody) through analytical methods remains challenging due to the heterogeneity of drug conjugation as well as the numerous post-translational modifications possible in the monoclonal antibody. Herein, we report on the use of high-resolution ion mobility spectrometry separations in structures for lossless ion manipulations coupled to mass spectrometry (SLIM IMS-MS) for the rapid and simultaneous characterization of the drug load profile (i.e., stoichiometric distribution of the number of conjugated drugs present on the mAb), determination of the weighted average DAR in both the heavy and light chains of a model antibody-drug conjugate, and calculation of the overall DAR of the ADC. After chemical reduction of the ADC and a subsequent 31.5 m SLIM IMS separation, the various drug-bound antibody species could be well resolved for both chains. We also show signifi...Continue Reading

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Citations

Feb 17, 2021·Analytical Chemistry·Pengyi ZhaoHao Chen
Apr 10, 2021·Journal of the American Society for Mass Spectrometry·James R ArndtMelissa Sherman
Jul 12, 2021·Biochimica Et Biophysica Acta. Proteins and Proteomics·Kirsty SkeeneCris Lapthorn
May 23, 2020·Analytical Chemistry·Malin KällstenSara Bergström Lind

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