Advertisement

Selecting an Optimal Antibody for Antibody–Drug Conjugate Therapy: Internalization and Intracellular Localization

  • Jay Harper
  • Shenlan Mao
  • Patrick Strout
  • Adeela Kamal
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1045)

Abstract

Antibody–drug conjugates (ADCs) combine the selectivity of a monoclonal antibody with the killing potency of a cytotoxic drug. For an antibody to function as a successful component of an ADC, it needs to bind to the target antigen on the surface of tumor cells and then be internalized by the cell. Following internalization, the ADC has to be transported to the lysosome where subsequent intracellular processing of the ADC will release the biologically active drug to exert its cytotoxic effects on tumor cells. This chapter describes some of the techniques that are currently used to determine internalization and proper intracellular trafficking of antibodies in order to select an optimal antibody for ADC therapeutics.

Key words

Antibodies Antibody–drug conjugate Internalization Flow cytometry Confocal microscopy 

References

  1. 1.
    Ducry L, Stump B (2010) Antibody–drug conjugates: linking cytotoxic payloads to monoclonal antibodies. Bioconjug Chem 21:5–13PubMedCrossRefGoogle Scholar
  2. 2.
    Katz J, Janik JE, Younes A (2011) Brentuximab vedotin (SGN-35). Clin Cancer Res 17:6428–6436PubMedCrossRefGoogle Scholar
  3. 3.
    Verma S, Miles D, Gianni L, Krop IE, Welslau M, Baselga J, Pegram M, Oh DY, Dieras V, Guardino E, Fang L, Lu MW, Olsen S, Blackwell K (2012) Trastuzumab emtansine for HER2-positive advanced breast cancer. N Engl J Med 367:1783–1791PubMedCrossRefGoogle Scholar
  4. 4.
    Alley SC, Okeley NM, Senter PD (2010) Antibody–drug conjugates: targeted drug delivery for cancer. Curr Opin Chem Biol 14:529–537PubMedCrossRefGoogle Scholar
  5. 5.
    Polson AG, Calemine-Fanaux J, Chan P, Chang W, Christensen E, Clark S, de Sauvage FJ, Eaton D, Elkins K, Elliot JM, Frantz G, Fuji RN, Gray A, Harden K, Ingle GS, Kljavin NM, Koeppen H, Nelson C, Prabju S, Raab H, Ross S, Stephan J-P, Scales SJ, Spencer SD, Vandlen R, Wranik B, Yu S-F, Zheng B, Ebens A (2009) Antibody–drug conjugates for the treatment of non-Hodgkin’s lymphoma: target and linker-drug selection. Cancer Res 69:2358–2364PubMedCrossRefGoogle Scholar
  6. 6.
    Carter PJ, Senter PD (2008) Antibody–drug conjugates for cancer therapy. Cancer J 14:154–169PubMedCrossRefGoogle Scholar
  7. 7.
    Lewis Phillips GD, Li G, Dugger DL, Crocker LM, Parsons KL, Mai E, Blättler WA, Lambert JM, Chari RVJ, Lutz RJ, Wong WLT, Jacobson FS, Koeppen H, Schwall RH, Kenkare-Mitra SR, Spencer SD, Sliwkowski MX (2008) Targeting HER2-positive breast cancer with trastuzumab-DM1, and antibody-cytotoxic drug conjugate. Cancer Res 68:9280–9290PubMedCrossRefGoogle Scholar
  8. 8.
    Hommelgaard AM, Lerdrup M, van Deurs B (2004) Association with membrane protrusions makes ErbB2 an internalization-resistant receptor. Mol Biol Cell 15:1557–1567PubMedCrossRefGoogle Scholar
  9. 9.
    Rudnick SI, Lou J, Shaller CC, Tang Y, Klein-Szanto AJ, Weiner LM, Marks JD, Adams GP et al (2011) Influence of affinity and antigen internalization on the uptake and penetration of anti-HER2 antibodies in solid tumors. Cancer Res 71:2250–2259PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  • Jay Harper
    • 1
  • Shenlan Mao
    • 1
  • Patrick Strout
    • 1
  • Adeela Kamal
    • 1
  1. 1.Oncology Research, MedImmune Inc.GaithersburgUSA

Personalised recommendations