Separation window dependent multiple injection (SWDMI) for large scale analysis of therapeutic antibody  N -glycans

Kovács, Zsuzsanna; Szarka, Máté; Szigeti, Márton; Guttman, András

doi:10.1016/j.jpba.2016.06.002

Cited by 16 publications

(11 citation statements)

References 8 publications

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“…Along the same lines, a multiple injection approach for rapid large scale CE analysis followed by multicomponent optical detection with LIF was proposed. 180 As a proof of principle, the protocol was applied for rapid and large scale analysis of major monoclonal antibody (IgG) N-glycans. Nearly 100 samples were introduced in a single capillary, leading to a full analysis time of 4 h rather than 12 h when the conventional approach was applied.…”

Section: Applicationsmentioning

confidence: 99%

Capillary Electrophoresis: Trends and Recent Advances

et al. 2018

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Section: Applicationsmentioning

confidence: 99%

Capillary Electrophoresis: Trends and Recent Advances

et al. 2018

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“…Although, just a few molecules are naturally fluorescent which implies otherwise a derivatization step for the non-fluorescent analytes. Concerning the analysis of mAb glycosylations, chemical modification based on 8-Aminopyrene-1,3,6-trisulfonic-acid (APTS) reaction allows the derivation of fluorophoric group and the addition of three negative charges to the glycan moiety, which is considered today as one of the reference method [23,[25][26][27][28]. However recently, other dye reagents like 2-aminobenzoic acid (2-AA) (excitation: 325 nm emission: 405 nm) [29] and Teal™ (excitation and emission: same as APTS) [28], providing respectively one and three negative charges, have been reported for glycan analysis.…”

Section: Technical Considerationsmentioning

confidence: 99%

“…In 2016 Kovacs et al reported a high-throughput N-glycan analysis by CGE-LIF with APTS labeled glycans [25]. [27].…”

Section: Glycan Analysismentioning

confidence: 99%

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

Lechner

Giorgetti

Gahoual

et al. 2019

Journal of Chromatography B

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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.

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“… 27 − 31 Injections can even be multiplexed to deliver packets of sample within a single run to eliminate dwell times. 32 Innovations in instrumentation and in applications of the methodology continue and can be observed as a push to make the technique more accessible to researchers through commercially available solutions to interfacing CE with a mass spectrometer, as well as databases and software that increase the ease of data analysis. Analytical techniques are continually adapted to meet the growing challenges in carbohydrate analysis and CE plays a unique role in glycoscience research.…”

Section: Introductionmentioning

confidence: 99%

Capillary Electrophoresis Separations of Glycans

et al. 2018

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Capillary electrophoresis has emerged as a powerful approach for carbohydrate analyses since 2014. The method provides high resolution capable of separating carbohydrates by charge-to-size ratio. Principle applications are heavily focused on N-glycans, which are highly relevant to biological therapeutics and biomarker research. Advances in techniques used for N-glycan structural identification include migration time indexing and exoglycosidase and lectin profiling, as well as mass spectrometry. Capillary electrophoresis methods have been developed that are capable of separating glycans with the same monosaccharide sequence but different positional isomers, as well as determining whether monosaccharides composing a glycan are alpha or beta linked. Significant applications of capillary electrophoresis to the analyses of N-glycans in biomarker discovery and biological therapeutics are emphasized with a brief discussion included on carbohydrate analyses of glycosaminoglycans and mono-, di-, and oligosaccharides relevant to food and plant products. Innovative, emerging techniques in the field are highlighted and the future direction of the technology is projected based on the significant contributions of capillary electrophoresis to glycoscience from 2014 to the present as discussed in this review.

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Separation window dependent multiple injection (SWDMI) for large scale analysis of therapeutic antibody N -glycans

Cited by 16 publications

References 8 publications

Capillary Electrophoresis: Trends and Recent Advances

Capillary Electrophoresis: Trends and Recent Advances

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

Capillary Electrophoresis Separations of Glycans

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