A new anti-human Fc method to capture and analyze ADCs for characterization of drug distribution and the drug-to-antibody ratio in serum from pre-clinical species

Excoffier, Mélissa; Janin-Bussat, Marie-Claire; Beau‐Larvor, Charlotte; Troncy, Lysiane; Corvaı̈a, Nathalie; Beck, Alain; Klinguer‐Hamour, Christine

doi:10.1016/j.jchromb.2016.05.037

Cited by 14 publications

(6 citation statements)

References 25 publications

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“…Existing approaches to quantify the conjugated species of ADCs involve a ligand-binding assay (LBAs) for conjugated antibody or hybrid LBA/liquid chromatography/tandem-mass spectrometry (LC/MS/MS) for quantitation of conjugated drug. Now, multilevel LC-MS workflows are more frequently used both for 2G [197,198] and 3G ADCs [199,200] as well as for protein-drug conjugate (PDC, such as centyrin scaffolds [201,202], based on immune-enrichment [203,204] and high-resolution MS [205].…”

Section: Adc Bioanalyticsmentioning

confidence: 99%

Cutting-edge multi-level analytical and structural characterization of antibody-drug conjugates: present and future

Beck

D’Atri

Ehkirch

et al. 2019

Expert Review of Proteomics

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Introduction: The development and optimization of antibody drug conjugates (ADCs) rely on improving their analytical and bioanalytical characterization, by assessing critical quality attributes (CQAs). Among the CQAs, the glycoprofile, drug load distribution (DLD), the amount of unconjugated antibody (D0), the average drug-to-antibody ratio (DAR), the drug conjugation sites and the residual drug-linker and related product proportions (SMDs) in addition to high and low molecular weight species (H/LMWS) are the most important ones. Areas covered: The analytical and structural toolbox for the characterization of 1 st , 2 d and 3 d generation ADCs was significantly extended in the last 3 years. Here, we reviewed state-ofthe-art techniques, such as liquid chromatography, high resolution native and ion mobility mass spectrometry, multidimensional LC and capillary electrophoresis hyphenated to mass spectrometry, reported mainly since 2016. Expert commentary: These emerging techniques allow a deep insight into important CQAs that are related to ADC Chemistry Manufacturing and Control (CMC) as well as an improved understanding of in vitro and in vivo ADC biotransformations. This knowledge and the development of quantitative bioanalytical assays will continue to contribute to earlydevelopability assessment for the optimization of all the ADC components (i.e., antibody, drug, and linker) and help to bring next-generation candidate ADC into the clinic and hopefully to the market.

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Section: Adc Bioanalyticsmentioning

confidence: 99%

Cutting-edge multi-level analytical and structural characterization of antibody-drug conjugates: present and future

Beck

D’Atri

Ehkirch

et al. 2019

Expert Review of Proteomics

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“…60,61 In this report, the ADC was purified from real samples using the magnetic beads with an immobilized antibody against monomethyl auristatin E (MMAE), which is a low-molecularweight drug moiety of the ADC, and directly separated by reverse-phase LC using a polymer capillary column (0.3 × 50 mm), followed by the final quantification by ESI-q-TOFMS. The ADC is quantified by its original molecular weight, which was calculated by the deconvolution of the multiple charged ions generated by the ESI.…”

Section: Bioanalysis Of Therapeutic Mabs and Adcs As Use Of Intact Prmentioning

confidence: 99%

Current Mass Spectrometric Tools for the Bioanalyses of Therapeutic Monoclonal Antibodies and Antibody-Drug Conjugates

et al. 2018

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The increase in the use of therapeutic monoclonal antibodies (mAbs) and antibody-drug conjugates (ADCs) has made the detailed bioanalysis of these drugs essential not only for planning optimal therapeutic programs for clinical practice, but also for evaluating the biological equivalencies in the development of other biosimilars. The ligand binding assays that are widely in use now are being replaced rapidly by the highly accurate, sensitive, and selective analytical method using a mass spectrometer. This review will discuss the progress in and challenges observed during the development of a mass spectrometry-based bioanalytical method for therapeutic mAbs and ADCs.

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“…The reduced and optimized sample preparation time also minimized assay artifacts that resulted from ex vivo drug metabolism. Excoffier et al [14] reported a new anti-human Fc method to capture and analyze ADCs for characterization of drug distribution and the drug-to-antibody ratio (DAR) in serum from pre-clinical species. The method profiled ADC DAR distribution with inter-chain disulfide bonds reduced after anti-human Fc capture.…”

Section: Dar Distribution-intact Adc Measurementmentioning

confidence: 99%

Current status of antibody-drug conjugate bioanalysis

Wang¹

2017

J Appl Bioanal

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Keywords: antibody-drug conjugate bioanalysis, ligand-binding assay, hybrid ligand-binding/LC-MS assay, drug-toantibody ratio Antibody-drug conjugate (ADC) consists of a cytotoxic drug covalently bound to a monoclonal antibody via a linker. Because of the complexity of ADC structure unique bioanalytical strategies are needed to identify, characterize and quantify the ADC species most relevant to safety and efficacy. ADC bioanalysis need an integrated bioanalytical approach including ligand-binding assay (LBA) and LC-MS based assays to provide comprehensive characterization of the exposure/response relationship. This mini-review will summarize recent publications on ADC bioanalytical strategies and will focus on the publications within last three years on the advancement of hybrid ligand-binding/LC-MS methods for ADC PK and stability evaluation and for intact ADC-DAR distribution measurement to profile ADC biotransformation and catabolism. Special attention is paid to the publications on selection of ADC analytes, assay platforms and DAR characteristics of LBA as well as on the transition of the bioanalytical testing strategy at different phases of clinical development.

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A new anti-human Fc method to capture and analyze ADCs for characterization of drug distribution and the drug-to-antibody ratio in serum from pre-clinical species

Cited by 14 publications

References 25 publications

Cutting-edge multi-level analytical and structural characterization of antibody-drug conjugates: present and future

Cutting-edge multi-level analytical and structural characterization of antibody-drug conjugates: present and future

Current Mass Spectrometric Tools for the Bioanalyses of Therapeutic Monoclonal Antibodies and Antibody-Drug Conjugates

Current status of antibody-drug conjugate bioanalysis

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