Characterization of ADCs by Capillary Electrophoresis

  • Wenjing Ning
  • Yanqun ZhaoEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2078)


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.

Key words

Antibody-drug conjugates CE Reduced and non-reduced CE-SDS iCIEF Charge heterogeneity 


  1. 1.
    Diamantis N, Banerji U (2016) Antibody-drug conjugates—an emerging class of cancer treatment. Br J Cancer 114:362. Scholar
  2. 2.
    Beck A, Reichert JM (2014) Antibody-drug conjugates. MAbs 6(1):15–17. Scholar
  3. 3.
    McCombs JR, Owen SC (2015) Antibody drug conjugates: design and selection of linker, payload and conjugation chemistry. AAPS J 17(2):339–351. Scholar
  4. 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. Scholar
  5. 5.
    Chudasama V, Maruani A, Caddick S (2016) Recent advances in the construction of antibody–drug conjugates. Nat Chem 8:114. Scholar
  6. 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. Scholar
  7. 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. Scholar
  8. 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. Scholar
  9. 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. Scholar
  10. 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. Scholar
  11. 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. Scholar
  12. 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. Scholar
  13. 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. Scholar
  14. 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. Scholar
  15. 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. Scholar

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

Authors and Affiliations

  1. 1.Process Analytical ChemistryAbbVie Inc.North ChicagoUSA

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