Abstract
The development of new therapeutic monoclonal antibodies (mAbs) for cancer therapy will rise in the following years. The evaluation of biological activity of mAbs is required during drug development and during drug production as quality control test. Antibody-dependent cell-mediated cytotoxicity (ADCC) is a desirable activity of anticancer mAbs. Here, we describe a flow cytometry-based method to quantify ADCC that combines the staining of cancer cells, effector cells, and dead cells, with specific dyes. This method is inexpensive, has low background, and avoids the use of radioisotopes.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Scott AM, Wolchok JD, Old LJ (2012) Antibody therapy of cancer. Nat Rev Cancer 12:278–287
Adler MJ, Dimitrov DS (2012) Therapeutic antibodies against cancer. Hematol Oncol Clin North Am 26:447–481, vii
Harrison A, Liu Z, Makweche S et al (2012) Methods to measure the binding of therapeutic monoclonal antibodies to the human Fc receptor FcgammaRIII (CD16) using real time kinetic analysis and flow cytometry. J Pharm Biomed Anal 63:23–28
Levy EM, Roberti MP, Mordoh J (2011) Natural killer cells in human cancer: from biological functions to clinical applications. J Biomed Biotechnol 2011:676198
Derby E, Reddy V, Kopp W et al (2001) Three-color flow cytometric assay for the study of the mechanisms of cell-mediated cytotoxicity. Immunol Lett 78:35–39
Collins DM, O'Donovan N, McGowan PM et al (2012) Trastuzumab induces antibody-dependent cell-mediated cytotoxicity (ADCC) in HER-2-non-amplified breast cancer cell lines. Ann Oncol 23:1788–1795
Yeo C, Saunders N, Locca D et al (2009) Ficoll-Paque versus Lymphoprep: a comparative study of two density gradient media for therapeutic bone marrow mononuclear cell preparations. Regen Med 4:689–696
Pollara J, Hart L, Brewer F et al (2011) High-throughput quantitative analysis of HIV-1 and SIV-specific ADCC-mediating antibody responses. Cytometry A 79:603–612
Green MR, Kennell AS, Larche MJ et al (2005) Natural killer cell activity in families of patients with systemic lupus erythematosus: demonstration of a killing defect in patients. Clin Exp Immunol 141:165–173
Acknowledgments
We thank Dr. Frank Robledo-Avila and Dra. Mayra Pérez-Tapia for the critical review of this manuscript. Assays presented in this work were performed at “Unidad de Desarrollo e Investigación en Bioprocesos (UDIBI), Instituto Politécnico Nacional.” The work is supported by a grant from “Instituto Científico Pfizer, México” and grants IN219613 (PAPIIT-UNAM) and 225313 (Conacyt) to MAV-V.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Science+Business Media New York
About this protocol
Cite this protocol
Salinas-Jazmín, N., Hisaki-Itaya, E., Velasco-Velázquez, M.A. (2014). A Flow Cytometry-Based Assay for the Evaluation of Antibody-Dependent Cell-Mediated Cytotoxicity (ADCC) in Cancer Cells. In: Robles-Flores, M. (eds) Cancer Cell Signaling. Methods in Molecular Biology, vol 1165. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0856-1_16
Download citation
DOI: https://doi.org/10.1007/978-1-4939-0856-1_16
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-0855-4
Online ISBN: 978-1-4939-0856-1
eBook Packages: Springer Protocols