Skip to main content
SpringerLink
Log in

Characterization of ADCs by Capillary Electrophoresis

  • Protocol
  • First Online:
Antibody-Drug Conjugates

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2078))

Abstract

Capillary electrophoresis (CE) is a highly efficient separation technique that resolves ions based on their electrophoretic mobility in the presence of an applied voltage. It has been broadly applied for characterizing biotherapeutics including ADCs. In this chapter, step-by-step procedures for characterizing ADCs using CE will be described with focus placed on reduced and non-reduced capillary electrophoresis sodium dodecyl sulfate (CE-SDS) for purity determination and imaged capillary isoelectric focusing (iCIEF) for charge heterogeneity analysis.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 99.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 129.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 199.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Diamantis N, Banerji U (2016) Antibody-drug conjugates—an emerging class of cancer treatment. Br J Cancer 114:362. https://doi.org/10.1038/bjc.2015.435. https://www.nature.com/articles/bjc2015435#supplementary-information

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Beck A, Reichert JM (2014) Antibody-drug conjugates. MAbs 6(1):15–17. https://doi.org/10.4161/mabs.27436

    Article  PubMed  Google Scholar 

  3. McCombs JR, Owen SC (2015) Antibody drug conjugates: design and selection of linker, payload and conjugation chemistry. AAPS J 17(2):339–351. https://doi.org/10.1208/s12248-014-9710-8

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Beck A, Goetsch L, Dumontet C, Corvaïa N (2017) Strategies and challenges for the next generation of antibody–drug conjugates. Nat Rev Drug Discov 16:315. https://doi.org/10.1038/nrd.2016.268

    Article  CAS  PubMed  Google Scholar 

  5. Chudasama V, Maruani A, Caddick S (2016) Recent advances in the construction of antibody–drug conjugates. Nat Chem 8:114. https://doi.org/10.1038/nchem.2415

    Article  CAS  PubMed  Google Scholar 

  6. Agarwal P, Bertozzi CR (2015) Site-specific antibody–drug conjugates: the Nexus of bioorthogonal chemistry, protein engineering, and drug development. Bioconjug Chem 26(2):176–192. https://doi.org/10.1021/bc5004982

    Article  CAS  PubMed  Google Scholar 

  7. Panowski S, Bhakta S, Raab H, Polakis P, Junutula JR (2014) Site-specific antibody drug conjugates for cancer therapy. MAbs 6(1):34–45. https://doi.org/10.4161/mabs.27022

    Article  PubMed  Google Scholar 

  8. Chen T, Chen Y, Stella C, Medley CD, Gruenhagen JA, Zhang K (2016) Antibody-drug conjugate characterization by chromatographic and electrophoretic techniques. J Chromatogr B 1032:39–50. https://doi.org/10.1016/j.jchromb.2016.07.023

    Article  CAS  Google Scholar 

  9. Gahoual R, Giorgetti J, Beck A, Leize-Wagner E, François Y-N (2018) Chapter 19 - biopharmaceutical applications of capillary Electromigration methods. In: Poole CF (ed) Capillary electromigration separation methods. Elsevier, Amsterdam, pp 453–480. https://doi.org/10.1016/B978-0-12-809375-7.00021-6

    Chapter  Google Scholar 

  10. Dai J, Lamp J, Xia Q, Zhang Y (2018) Capillary isoelectric focusing-mass spectrometry method for the separation and online characterization of intact monoclonal antibody charge variants. Anal Chem 90(3):2246–2254. https://doi.org/10.1021/acs.analchem.7b04608

    Article  CAS  PubMed  Google Scholar 

  11. Gahoual R, Beck A, Leize-Wagner E, François Y-N (2016) Cutting-edge capillary electrophoresis characterization of monoclonal antibodies and related products. J Chromatogr B 1032:61–78. https://doi.org/10.1016/j.jchromb.2016.05.028

    Article  CAS  Google Scholar 

  12. Redman EA, Mellors JS, Starkey JA, Ramsey JM (2016) Characterization of intact antibody drug conjugate variants using microfluidic capillary electrophoresis–mass spectrometry. Anal Chem 88(4):2220–2226. https://doi.org/10.1021/acs.analchem.5b03866

    Article  CAS  PubMed  Google Scholar 

  13. Stolz A, Jooß K, Höcker O, Römer J, Schlecht J, Neusüss C (2018) Recent advances in capillary electrophoresis-mass spectrometry: Instrumentation, methodology and applications. Electrophoresis 40(1):79–112. https://doi.org/10.1002/elps.201800331

    Article  CAS  PubMed  Google Scholar 

  14. Wiggins B, Liu-Shin L, Yamaguchi H, Ratnaswamy G (2015) Characterization of cysteine-linked conjugation profiles of immunoglobulin G1 and immunoglobulin G2 antibody–drug conjugates. J Pharm Sci 104(4):1362–1372. https://doi.org/10.1002/jps.24338

    Article  CAS  PubMed  Google Scholar 

  15. Valliere-Douglass JF, Lewis P, Salas-Solano O, Jiang S (2015) Solid-state mAbs and ADCs subjected to heat-stress stability conditions can be covalently modified with buffer and excipient molecules. J Pharm Sci 104(2):652–665. https://doi.org/10.1002/jps.24276

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Process Analytical Chemistry, AbbVie Inc., North Chicago, IL, USA

    Wenjing Ning & Yanqun Zhao

Authors
  1. Wenjing Ning
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yanqun Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yanqun Zhao .

Editor information

Editors and Affiliations

  1. Department of Pharmacy and Pharmaceutical Sciences, Binghamton University, Binghamton, NY, USA

    L. Nathan Tumey

Rights and permissions

Reprints and permissions

Copyright information

© 2020 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Ning, W., Zhao, Y. (2020). Characterization of ADCs by Capillary Electrophoresis. In: Tumey, L. (eds) Antibody-Drug Conjugates. Methods in Molecular Biology, vol 2078. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9929-3_17

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-9929-3_17

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9928-6

  • Online ISBN: 978-1-4939-9929-3

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation