Determination of ADC Concentration by Ligand-Binding Assays.

Methods Mol Biol

Department of Pharmaceutical Sciences, School of Pharmacy and Pharmaceutical Sciences, The State University of New York, University at Buffalo, Buffalo, NY, USA.

Published: January 2021

AI Article Synopsis

  • Total antibodies, conjugated antibodies or antibody-conjugated drugs, and free drugs are essential for understanding exposure-response in antibody-drug conjugates (ADCs).
  • Bioanalytical methods like ligand-binding assays (LBA) and LC-MS/MS are employed to analyze these components.
  • The text outlines a methodology for developing a solid-phase ELISA, which is commonly used to measure total antibody levels in biological samples, and can be adjusted to quantify conjugated antibody levels as well.

Article Abstract

Total antibody, conjugated antibody or antibody-conjugated drug, and free drug are key analytes required to establish exposure-response relationships for ADCs. Therefore, bioanalytical strategies for ADCs include ligand-binding assays (LBA) and LC-MS/MS methods. Here we describe detailed methodology to develop a solid-phase-based enzyme-linked immunosorbent assay (ELISA), which is the most widely used LBA to quantify large-molecule components of ADC in biological matrices such as plasma, serum, tumor, or tissue homogenates. The approach presented here is designed to quantify total antibody concentrations in ADC containing samples, and can be easily adapted to quantify conjugated antibody concentrations.

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http://dx.doi.org/10.1007/978-1-4939-9929-3_26DOI Listing

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