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Fast Afucosylation Profiling of Glycoengineered Antibody Subunits by Middle-Up Mass Spectrometry

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Mass Spectrometry of Glycoproteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2271))

Abstract

Middle-up LC-MS antibody characterization workflows using reduction or IdeS digestion for a focused assessment of N-glycan profiling of three representative glycoengineered monoclonal antibodies (mAbs), namely, obinutuzumab (GlycomAb technology, Glycart/Roche), benralizumab (Potelligent Technology, BioWa, Kyowa Kirin) and mAb B (kifunensine) and compared to mAb A, produced in a common CHO cell line. In addition, EndoS or EndoS2 enzyme are used for quantitative determination of Fc-glycan core afucosylation and high mannose for these antibodies, as requested by health authorities for Fc-competent therapeutics mAbs critical quality attributes (CQAs).

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Author information

Authors and Affiliations

  1. Pierre Fabre Laboratories, IRPF—Centre d’Immunologie Pierre-Fabre (CIPF), Saint-Julien-en-Genevois, France

    Elsa Wagner-Rousset, Olivier Colas, Stéphane Chenu & Alain Beck

  2. Laboratoire de Spectrométrie de Masse des Interactions et des Systèmes (LSMIS), UMR 7140, Université de Strasbourg, CNRS, Strasbourg, France

    Yannis-Nicolas François

  3. School of Pharmaceutical Sciences, University of Geneva, CMU, Geneva, Switzerland

    Davy Guillarme

  4. Laboratoire de Spectrométrie de Masse BioOrganique, IPHC UMR 7178, Université de Strasbourg, CNRS, Strasbourg, France

    Sarah Cianferani

  5. Spectroswiss Sarl, EPFL Innovation Park, Lausanne, Switzerland

    Yury O. Tsybin

  6. Genovis AB, Lund, Sweden

    Jonathan Sjögren

  7. Quality Assistance SA, Donstiennes, Belgium

    Arnaud Delobel

  8. Institute of Pharmaceutical Sciences of Western Switzerland (ISPSO), University of Geneva, CMU, Geneva, Switzerland

    Davy Guillarme

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  1. Elsa Wagner-Rousset
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  2. Olivier Colas
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  3. Stéphane Chenu
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  4. Yannis-Nicolas François
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  7. Yury O. Tsybin
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  8. Jonathan Sjögren
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  9. Arnaud Delobel
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  10. Alain Beck
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Corresponding author

Correspondence to Alain Beck .

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  1. Quality Assistance S.A., Donstiennes, Belgium

    Arnaud Delobel

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Wagner-Rousset, E. et al. (2021). Fast Afucosylation Profiling of Glycoengineered Antibody Subunits by Middle-Up Mass Spectrometry. In: Delobel, A. (eds) Mass Spectrometry of Glycoproteins. Methods in Molecular Biology, vol 2271. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1241-5_5

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  • DOI: https://doi.org/10.1007/978-1-0716-1241-5_5

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1240-8

  • Online ISBN: 978-1-0716-1241-5

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