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Assessment of Macro- and Microheterogeneity of Monoclonal Antibodies Using Capillary Zone Electrophoresis Hyphenated with Mass Spectrometry

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Capillary Electrophoresis-Mass Spectrometry

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2531))

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Abstract

This chapter focuses on the application of capillary zone electrophoresis hyphenated with mass spectrometry (CZE-MS) for the characterization of monoclonal antibodies (mAbs). mAbs are complex molecules comprising different glycoforms and many other posttranslational modifications. In addition to this inherent microheterogeneity, misassembling of antibodies can take place during production contributing to their macroheterogeneity. CZE-MS is a versatile and powerful technique which has demonstrated high potential for the assessment of both micro- and macroheterogeneity of mAbs. In this chapter, technical and practical considerations for the characterization of mAbs by CZE-MS are described. CE-MS interfacing, capillary coatings for the prevention of mAb adsorption, and sample preparation considerations are covered in detail. The assessment of the macro- and microheterogeneity is discussed and exemplified through three different approaches involving analysis of intact, enzymatically digested, and reduced antibodies. The examples also illustrate the use of two commercially available interfacing techniques (i.e., sheath liquid and sheathless) as well as different types of capillary coatings (positively charged and neutral coatings).

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Authors and Affiliations

  1. Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, The Netherlands

    Christoph Gstöttner, Manfred Wuhrer & Elena Domínguez-Vega

  2. Division of BioAnalytical Chemistry, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands

    Rob Haselberg & Govert W. Somsen

Authors
  1. Christoph Gstöttner
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  2. Rob Haselberg
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  3. Manfred Wuhrer
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  4. Govert W. Somsen
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  5. Elena Domínguez-Vega
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Corresponding author

Correspondence to Elena Domínguez-Vega .

Editor information

Editors and Affiliations

  1. Aalen University, Aalen, Baden-Württemberg, Germany

    Christian Neusüß

  2. Aalen University, Aalen, Baden-Württemberg, Germany

    Kevin Jooß

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Gstöttner, C., Haselberg, R., Wuhrer, M., Somsen, G.W., Domínguez-Vega, E. (2022). Assessment of Macro- and Microheterogeneity of Monoclonal Antibodies Using Capillary Zone Electrophoresis Hyphenated with Mass Spectrometry. In: Neusüß, C., Jooß, K. (eds) Capillary Electrophoresis-Mass Spectrometry . Methods in Molecular Biology, vol 2531. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2493-7_9

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  • DOI: https://doi.org/10.1007/978-1-0716-2493-7_9

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2492-0

  • Online ISBN: 978-1-0716-2493-7

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