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Drug Removal Strategies in Competitive Ligand Binding Neutralizing Antibody (NAb) Assays: Highly Drug-Tolerant Methods and Interpreting Immunogenicity Data

Abstract

Neutralizing anti-drug antibody (NAb) assays often have lower drug tolerance (DT) than trough drug concentrations, potentially under-estimating NAb incidence. To improve DT, drug-specific proteins were coupled to magnetic beads to deplete drug in the sample. To avoid interference from carryover, drug-specific proteins that did not interfere in the NAb assay, such as target or non-blocking anti-drug antibodies, were selected. With the drug depletion step, DT improved by > 10-fold in two competitive ligand binding NAb assays. Analysis of anti-drug antibody positive clinical samples with elevated drug levels demonstrated that NAb incidence was under-estimated without the drug depletion step. However, these NAb-positive samples had low titer and no impact on drug concentrations.

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Acknowledgments

We are extremely grateful to Kevin Laurino, Stephen Okaine, Eileen Ceconi, Colin Stefan, Lisa DeStefano, and Matt Andisik from the Clinical Bioanalysis group in the Bioanalytical Sciences Department at Regeneron for generating drug concentration data.

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Correspondence to Michael A. Partridge.

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Partridge, M.A., Karayusuf, E.K., Shyu, G. et al. Drug Removal Strategies in Competitive Ligand Binding Neutralizing Antibody (NAb) Assays: Highly Drug-Tolerant Methods and Interpreting Immunogenicity Data. AAPS J 22, 112 (2020). https://doi.org/10.1208/s12248-020-00497-2

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KEY WORDS

  • competitive ligand binding assays
  • drug depletion
  • drug tolerance
  • immunogenicity
  • neutralizing antibody