Antibody Dependent Cellular Cytotoxicity (ADCC)

Antibody-dependent cellular cytotoxicity (ADCC), also referred to as antibody-dependent cell-mediated cytotoxicity, is a mechanism of cell-mediated immune defence whereby an effector cell of the immune system actively lyses a target cell, whose membrane-surface antigens have been bound by specific antibodies. It is one of the mechanisms through which antibodies, as part of the humoral immune response, can act to limit and contain infection.

  • Examine ADCC activity of your proprietary therapeutic antibody using physiologically relevant in vitro model 
  • Bespoke ADCC assays using target and effector cells (PBMCs, T-cells, NK cells) of your choice
  • Develop cell lines expressing target antigen of your interest 
 
Our scientific team can propose several different approaches – 
1. Transfecting target cells with GFP and measuring the loss of intracellular GFP  following incubation with effector cells and therapeutic test antibody as a surrogate for cytotoxicity. This model provides you the flexibility to work with different combinations of target/effector cells and test antibodies and can be  applied for validation projects
2. Using a reporter cell line – Jurkat cell line (effector cells) stably expressing human FcγRIIIa V158 and NFAT-induced luciferase. Following engagement with the Fc region of a relevant antibody bound to a Target Cell, effector Cells expressing FcγRIIIa transduce intracellular signals resulting in NFAT-mediated  luciferase activity that can be easily quantified. 
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Antibody-dependent cellular cytotoxicity (ADCC)

Raji cells were treated with Anti-CD20 antibody and T cells and the Relative Luminescence Units (RLU) were measured 6 hours [A] and 24 hours [B] post incubation. The higher concentration of the Anti-CD20 resulted in an increase in the activation of the NFAT pathway. This corresponds directly to an increase in the target cell death.

Assessment of ADCC via GFP release

Incubation of GFP labelled MCF7 (target) cells with both Trastuzumab and PMA activated Thp1 (effector) cells induces a loss of intracellular GFP suggesting an increased antibody-dependent cytotoxic effect on MCF7

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