High resolution separations of charge variants and disulfide isomers of monoclonal antibodies and antibody drug conjugates using ultra-high voltage capillary electrophoresis with high electric field strength

Henley, W. Hampton; He, Yue; Mellors, J. Scott; Batz, Nicholas G.; Ramsey, J. Michael; Jorgenson, James W.

doi:10.1016/j.chroma.2017.07.059

Cited by 17 publications

(6 citation statements)

References 27 publications

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“…Recently, Henley et al successfully demonstrated application of UHV-CE in separation of mAb charge variants, drug conjugates and disulfide isomers with better resolution and short analysis time. 124…”

Section: Ultra-high Voltage Capillary Electrophoresismentioning

confidence: 99%

Challenges and new frontiers in analytical characterization of antibody-drug conjugates

Wagh

Song

Zeng

et al. 2018

mAbs

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Antibody-drug conjugates (ADCs) are a growing class of biotherapeutics in which a potent small molecule is linked to an antibody. ADCs are highly complex and structurally heterogeneous, typically containing numerous product-related species. One of the most impactful steps in ADC development is the identification of critical quality attributes to determine product characteristics that may affect safety and efficacy. However, due to the additional complexity of ADCs relative to the parent antibodies, establishing a solid understanding of the major quality attributes and determining their criticality are a major undertaking in ADC development. Here, we review the development challenges, especially for reliable detection of quality attributes, citing literature and new data from our laboratories, highlight recent improvements in major analytical techniques for ADC characterization and control, and discuss newer techniques, such as two-dimensional liquid chromatography, that have potential to be included in analytical control strategies.

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Section: Ultra-high Voltage Capillary Electrophoresismentioning

confidence: 99%

Challenges and new frontiers in analytical characterization of antibody-drug conjugates

Wagh

Song

Zeng

et al. 2018

mAbs

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“…The analysis has been made with a 120 kV electric field (2 kV/cm) in a HPMC coated capillary. This system enhanced resolution and separation speed allowing distinction of disulfide isomers [89]. More recently, CZE has also been developed to get the best repeatability in particular with one of the key source of differentiability of…”

Section: Technical Considerationsmentioning

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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“…Henley et al. applied ultra‐high voltage CE for the analysis of mAb drug conjugates, charge variants, and disulfide isomers. Voltages of more than 120 kV and electric field strength up to 2 kV/cm could be applied with the utilized instrument, refined from .…”

Section: Capillary Zone Electrophoresismentioning

confidence: 99%

Determination of protein charge variants with (imaged) capillary isoelectric focusing and capillary zone electrophoresis

Kahle

Wätzig

2018

Electrophoresis

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Biopharmaceuticals are increasingly present in pharmaceutical therapy. These highly complex molecules are still challenging for development and quality control. Charge heterogeneity is next to size heterogeneity, glycan analysis, or peptide mapping one main aspect of their thorough characterization. This review focuses on the charge-based analysis of biopharmaceuticals through CE. An overview is given about the three separation techniques cIEF, imaged cIEF and CZE for the analyses of charge variants, emphasizing assays and their precision. Especially the developments in the experimental conditions have been given a high priority: sample preparation, capillary properties, anolytes and catholytes, carrier ampholytes, sacrificing agents, solubility enhancement, isoelectric point marker, mobilization techniques, and detection modes have been discussed. CIEF shows a particularly high heterogeneity in its methods. Therefore, a design space is suggested that states the most important adjustable parameters and their range. This enables an adequate design of experiments for individual sample separations, in order to accelerate and simplify method developments.

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High resolution separations of charge variants and disulfide isomers of monoclonal antibodies and antibody drug conjugates using ultra-high voltage capillary electrophoresis with high electric field strength

Cited by 17 publications

References 27 publications

Challenges and new frontiers in analytical characterization of antibody-drug conjugates

Challenges and new frontiers in analytical characterization of antibody-drug conjugates

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

Determination of protein charge variants with (imaged) capillary isoelectric focusing and capillary zone electrophoresis

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