Insights from capillary electrophoresis approaches for characterization of monoclonal antibodies and antibody drug conjugates in the period 2016–2018

Abstract

Monoclonal antibodies (mAbs) and their related products as antibody-drug-conjugates (ADCs) or biosimilars represent a constantly growing class of molecules therapeutic proteins used as treatment against numerous diseases. These compounds can undergo several modifications which could alter the efficiency of treatments. In this context, several analytical methods were designed to deliver a comprehensive structural characterization and guarantee the quality of biotherapeutics. Capillary electrophoresis (CE) is considered today as a major technique for the analysis of biotherapeutics due to benefic characteristics as high resolution separation and miniaturized format. Different CE modes have been developed to characterize mAbs at different levels such as capillary gel electrophoresis (CGE), capillary isoelectric focusing (cIEF), and capillary zone electrophoresis (CZE). Recent developments in CE-mass spectrometry (MS) coupling assessed this technology as a promising tool to obtain high level structural characterization of biopharmaceuticals. Moreover, upcoming techniques such as 2D CE-MS and microfluidic systems are now emerging to offer new possibilities beyond actual limits. This review will be dedicated to discuss the state-of-the-art CE-based methods for the characterization of mAbs and ADCs in the period 2016-2018.

Combination of intact, middle-up and bottom-up levels to characterize 7 therapeutic monoclonal antibodies by capillary electrophoresis – Mass spectrometry

“…Regarding capillary zone electrophoresis (CZE)-based methods for the characterization of mAbs charges variants, until 2016, almost all literature described quality control application in terms of product heterogeneity using CZE with UV detection [19,21]. However, strategies with optical detection didn't allow to obtain structural information for the basic and/or acidic variants representing the limitation of these methodologies.…”

“…Traditional mAb characterization has been operated following a "bottom-up" strategy with a LC-MS/MS coupling or more recently with a sheathless CE-ESI-MS/MS coupling [9,19]. Although the important ability to identify different proteoforms with minor micro-heterogeneities, this approach can occur some artifactual modifications due to sample preparation and chemical treatment.…”

Antibody Conjugates-Recent Advances and Future Innovations

“…Over the last decade, significant advancements in analytical methods have been made to characterize ADCs and have been extensively reviewed [151][152][153][154]. These methods have focused on the major ADC attributes such as DAR, drug load distribution, residual linker-payload and related impurity levels, in addition to typical attributes for antibodies such as aggregation level, charge variants, and host cell protein level.…”

Progress in understanding COVID-19: insights from the omics approach

“…However, due to the complexity of neutralizing antibody (NAb), identifying the structural and molecular characteristics of NAb could accelerate vaccine design and drug discovery. MS-based methods provide an effective approach for quantifying therapeutic antibodies along with their modification forms, such as post-translational modifications [68][69][70][71]. To date, there are no reports on NAb analysis using MS, which remains challenging; however, we believe that MS can significantly improve NAb research.…”