Sensitive ELISA Method for the Measurement of Catabolites of Antibody-Drug Conjugates (ADCs) in Target Cancer Cells

Molecular Pharmaceutics
Paulin L Salomon, Rajeeva Singh

Abstract

A new, sensitive ELISA method has been developed which measures catabolites in cells and media upon processing of antibody-drug conjugates (ADCs) by target cancer cells. This ELISA method, exemplified for maytansinoid ADCs, uses competitive inhibition by a maytansinoid analyte of the binding of biotinylated antimaytansine antibody to an immobilized BSA-maytansinoid conjugate. Synthetic standards of several maytansinoid catabolites derived from ADCs with different linkers were tested and showed similar inhibition curves, with an EC50 of about 0.1 nM (0.03 pmol in an assay volume of 0.25 mL). This high sensitivity allowed quantification of catabolites from a methanolic cell extract and from the medium, generated from an ADC in 1 day using only about 1 million cells. The processing of anti-EpCAM and anti-CanAg ADCs with noncleavable linker (SMCC-DM1), disulfide linker (SPDB-DM4), and charged sulfonate-bearing disulfide linker (sulfo-SPDB-DM4), each containing an average of about four maytansinoid molecules per antibody, were compared in colon cancer cell lines (COLO 205 and HT-29). An 8-10-fold higher total level of catabolite was observed for anti-CanAg ADCs than for anti-EpCAM ADCs upon processing by COLO 205 cells, consistent w...Continue Reading

References

Jan 5, 2000·Biophysical Journal·Y E KorchevC R Edwards
Jun 5, 2003·Nature Biotechnology·Svetlana O DoroninaPeter D Senter
Jul 22, 2005·Clinical Cancer Research : an Official Journal of the American Association for Cancer Research·Puja SapraDavid M Goldenberg
Jul 11, 2006·Journal of Medicinal Chemistry·Wayne C WiddisonRavi V J Chari
Sep 24, 2008·Clinical Cancer Research : an Official Journal of the American Association for Cancer Research·Ezogelin OflazogluHans-Peter Gerber
Oct 1, 2009·Clinical Cancer Research : an Official Journal of the American Association for Cancer Research·Serengulam V GovindanDavid M Goldenberg
Nov 21, 2009·BMC Cancer·Sabine DenzelOlivier Gires
Jan 21, 2010·Clinical Cancer Research : an Official Journal of the American Association for Cancer Research·Nicole M OkeleyStephen C Alley
Mar 4, 2010·Cancer Research·Yelena V KovtunRavi V J Chari
Mar 24, 2010·Journal of Clinical Oncology : Official Journal of the American Society of Clinical Oncology·Anjali AdvaniLuis Fayad
Oct 13, 2010·Molecular Cancer Therapeutics·Emin OroudjevMary Ann Jordan
Mar 5, 2011·Clinical Cancer Research : an Official Journal of the American Association for Cancer Research·Thomas M CardilloDavid M Goldenberg
Apr 27, 2011·Journal of Medicinal Chemistry·Robert Y ZhaoRavi V J Chari
Oct 18, 2011·Clinical Cancer Research : an Official Journal of the American Association for Cancer Research·Jessica KatzAnas Younes
Dec 10, 2013·Hematology·Jonathan G Drachman, Peter D Senter
Mar 29, 2014·Angewandte Chemie·Ravi V J ChariWayne C Widdison
Jun 27, 2014·Journal of Medicinal Chemistry·John M Lambert, Ravi V J Chari
Aug 13, 2014·Proceedings of the National Academy of Sciences of the United States of America·Andrea E ProtaMichel O Steinmetz

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Citations

Oct 18, 2016·Journal of Pharmacological and Toxicological Methods·Malachy C UgwuRemigius U Agu
Jul 24, 2020·Expert Opinion on Investigational Drugs·Estelle Bourbon, Gilles Salles
Apr 12, 2018·Current Medicinal Chemistry·Nagula ShankaraiahLakshmi Soukya P S
Mar 18, 2017·Nature Reviews. Drug Discovery·Alain BeckNathalie Corvaïa
Oct 28, 2019·Biochemistry·Qing ZhangXuechen Li
May 16, 2019·Bioconjugate Chemistry·Dana N ThornlowChristopher A Alabi

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