A Novel System for the Quantification of the ADCC Activity of Therapeutic Antibodies

J Immunol Res. 2017:2017:3908289. doi: 10.1155/2017/3908289. Epub 2017 Sep 27.


Novel ADCC effector cells expressing the V-variant or F-variant of FcγRIIIa (CD16a) and firefly luciferase under the control of a chimeric promoter incorporating recognition sequences for the principal transcription factors involved in FcγRIIIa signal transduction, together with novel target cells overexpressing a constant high level of the specific antigen recognized by rituximab, trastuzumab, cetuximab, infliximab, adalimumab, or etanercept, confer improved sensitivity, specificity, and dynamic range in an ADCC assay relative to effector cells expressing a NFAT-regulated reporter gene and wild-type target cells. The effector cells also contain a normalization gene rendering ADCC assays independent of cell number or serum matrix effects. The novel effector and target cells in a frozen thaw-and-use format exhibit low vial-to-vial and lot-to-lot variation in their performance characteristics reflected by CVs of 10% or less. Homologous control target cells in which the specific target gene has been invalidated by genome editing providing an ideal control and a means of correcting for nonspecific effects were observed with certain samples of human serum. The novel effector cells and target cells expressing noncleavable membrane-bound TNFα have been used to quantify ADCC activity in serum from patients with Crohn's disease treated with infliximab and to relate ADCC activity to drug levels.

MeSH terms

  • Antibody-Dependent Cell Cytotoxicity*
  • Antigens, CD20 / genetics*
  • Antigens, CD20 / immunology
  • Cetuximab / metabolism
  • Crohn Disease / immunology*
  • ErbB Receptors / genetics*
  • ErbB Receptors / immunology
  • Etanercept / metabolism
  • Genes, Reporter / genetics
  • HEK293 Cells
  • Humans
  • Immunologic Techniques / methods*
  • Infliximab / metabolism
  • Jurkat Cells
  • NFATC Transcription Factors / genetics*
  • Receptor, ErbB-2 / genetics*
  • Receptor, ErbB-2 / immunology
  • Receptors, IgG / genetics*
  • Receptors, IgG / immunology
  • Rituximab / metabolism
  • Signal Transduction
  • T-Lymphocytes / physiology*
  • Transgenes / genetics
  • Trastuzumab / metabolism
  • Tumor Necrosis Factor-alpha / genetics*
  • Tumor Necrosis Factor-alpha / immunology


  • Antigens, CD20
  • FCGR3A protein, human
  • NFATC Transcription Factors
  • Receptors, IgG
  • Tumor Necrosis Factor-alpha
  • Rituximab
  • Infliximab
  • EGFR protein, human
  • ERBB2 protein, human
  • ErbB Receptors
  • Receptor, ErbB-2
  • Etanercept
  • Trastuzumab
  • Cetuximab