Cutting-edge mass spectrometry methods for the multi-level structural characterization of antibody-drug conjugates
Résumé
Antibody drug conjugates (ADCs) are highly cytotoxic drugs covalently attached via conditionally stable linkers to monoclonal antibodies (mAbs) and are among the most promising next-generation empowered biologics for cancer treatment. ADCs are more complex than naked mAbs, as the heterogeneity of the conjugates adds to the inherent microvariability of the biomolecules. The development and optimization of ADCs rely on improving their analytical and bioanalytical characterization by assessing several critical quality attributes, namely the distribution and position of the drug, the amount of naked antibody, the average drug to antibody ratio, and the residual druglinker and related product proportions. Here brentuximab vedotin (Adcetris®) and trastuzumab emtansine (Kadcyla®), the first and gold-standard hinge-cysteine and lysine drug conjugates, respectively, were chosen to develop new mass spectrometry (MS) methods and to improve multiplelevel structural assessment protocols.
Mots clés
Expert Review of Proteomics Antibody drug conjugate
bioanalysis
brentuximab vedotin
drug to antibody ratio
IdeS
ion mobility MS
native MS
mass spectrometry
trastuzumab emtansine Abbreviation List MS
ESI
electrospray ionization
ADC
antibody-drug-conjugate
DAR
drug-to-antibody ratio
HDX
hydrogen/deuterium exchange
BV
T-DM1
trastuzumab emtansine
HOS
higher order structure
mAb
monoclonal antibody
HER2
human epidermal growth factor receptor 2
human immunoglobilin G
IgG
UHPLC
ultra-high performance liquid chromatography
MS/MS
tandem mass spectrometry
SEC
size exclusion chromatography
HIC
hydrophobic interaction chromatography
SMCC
succinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate
CQA
critical quality attribute
rpHPLC
reversed-phase high performance liquid chromatography
CE
capillary electrophoresis
CE-SDS
capillary electrophoresis-sodium dodecyl sulphate
IEF
Domaines
Chimie analytique
Origine : Fichiers produits par l'(les) auteur(s)