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Homogeneous Antibody–Drug Conjugates via Glycoengineering

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Bioconjugation

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2033))

Abstract

Conventional antibody–drug conjugates (ADCs) randomly assemble small-molecule drugs onto Lys or Cys residues of a tumor-targeting antibody, featured with heterogeneity in payload numbers and conjugation positions. Glycosite-specific ADCs (gsADCs) link payload drugs onto IgG Fc N-glycans with high homogeneity that facilitates structural optimization and quality control for ADC drug development. In this protocol, we report two strategies for preparation of homogeneous ADCs via chemoenzymatic glycoengineering. First, an azido-tagged unnatural N-glycan substrate is transferred onto Fc glycosites of a therapeutic antibody through Endo-S-catalyzed glycoremodeling, followed by click reaction with an alkyne-tagged payload drug to give a well-defined gsADC. In an alternative way, glycoengineering of antibody with a natural sialylated N-glycan and successive selective oxidation of sialic acid moieties using sodium periodate provided an aldehyde handle on the glycans for conjugation with an aminooxy-assembled payload. These two strategies both enable gsADCs with high homogeneity in their conjugation sites, payload numbers, and glycoforms, which are characterized of a single mass under mass-spectral detection.

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Acknowledgments

This work is supported by the National Natural Science Foundation of China (NNSFC, No. 21572244), the SIMM Institute Fund (CASIMM0120153004), and the “Personalized Medicines—Molecular Signature-based Drug Discovery and Development,” Strategic Priority Research Program of the Chinese Academy of Sciences, Grant No. XDA12020311.

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Authors and Affiliations

  1. CAS Key Laboratory of Receptor Research, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Pudong, Shanghai, China

    Feng Tang, Wei Shi & Wei Huang

  2. University of Chinese Academy of Sciences, Beijing, People’s Republic of China

    Feng Tang, Wei Shi & Wei Huang

Authors
  1. Feng Tang
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  2. Wei Shi
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  3. Wei Huang
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Correspondence to Wei Huang .

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Editors and Affiliations

  1. Cellular and Molecular Immunology Laboratory, Vrije Universiteit Brussel, Brussels, Belgium

    Sam Massa

  2. In Vivo Cellular and Molecular Imaging Laboratory (ICMI), Vrije Universiteit Brussel, Brussels, Belgium

    Nick Devoogdt

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Tang, F., Shi, W., Huang, W. (2019). Homogeneous Antibody–Drug Conjugates via Glycoengineering. In: Massa, S., Devoogdt, N. (eds) Bioconjugation. Methods in Molecular Biology, vol 2033. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9654-4_15

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  • DOI: https://doi.org/10.1007/978-1-4939-9654-4_15

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9653-7

  • Online ISBN: 978-1-4939-9654-4

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