Non-Competing Clones Overcome Therapeutic Antibody Interference in Flow Cytometry 2260752

Abstract

Abstract Introduction The use of therapeutic antibodies in both basic research and clinical settings can interfere with flow cytometric analysis by masking target epitopes. This interference poses a significant challenge for identifying key cell populations and accurately assessing target expression levels. Methods To address this issue, we identified and validated antibody clones that do not compete with key therapeutic antibodies for binding, enabling reliable flow cytometry analysis even in the presence of these biologics. Results Specifically, we demonstrate non-competing clones for major therapeutic targets including CD38, PD-1, CD22, HER-2, and EGFR. These RUO antibodies retained strong binding despite pre-treatment with up to 40-fold excess of key therapeutic antibodies. Additionally, we provide a curated list of literature-reported non-competing clones for other targets. Recognizing the utility of both competing and non-competing clones in receptor occupancy assays, we also present clones that are blocked by their cognate therapeutic antibodies. Conclusion These data will support investigators in selecting appropriate antibody reagents tailored to their specific research needs when studying therapeutic biologics. Funding Source n/a Topic Categories Tumor Immunology: Checkpoints, Prevention, and Treatment (TIPT)