Volume 5 Supplement 8

22nd European Society for Animal Cell Technology (ESACT) Meeting on Cell Based Technologies

Open Access

ADCC potency assay: increased standardization with modified lymphocytes

BMC Proceedings20115(Suppl 8):P63

DOI: 10.1186/1753-6561-5-S8-P63

Published: 22 November 2011

For few years now, the use of monoclonal antibodies represents a significant progress in different therapeutic applications. In addition to commercialization of new products, important efforts in research and development have been made to launch new therapeutic antibodies.

Many antibodies act through a mechanism of Antibody-dependant cell cytotoxicity (ADCC). National Health Agencies recommend or require the use of biological activity assays (potency) in order to characterize those pharmaceutical products. The ADCC assay combines the 3 following elements:

  • The antibody of interest that is specific to a given antigen;

  • The targeted cells that express the antigen of interest at their surface;

  • The effector that can trigger the lysis the targeted cell when the antibody is linked to the antigen.

As the usually met ADCC assays use Natural Killer cells, isolated from healthy donors, as effectors they are hardly reproducible (Table 1). Thus, those assays can barely be validated when lots of pharmaceutical products are released. Furthermore, of the rare NK cell lines established in culture don’t express the CD16 receptor needed for the ADCC function.
Table 1

Advantages and drawbacks of ADCC effectors for the validation of a standardized ADCC assay

ADCC effector types



Primary NK or PBMC, isolated from donors

Representative from the genetic diversity

Not convenient for standardization

Requirement for donor genotyping

Requirement to evaluate several donors for accurate comparison

Exposure of operators to biohazard

Significant ressources required (budget and time) for cell preparation

Modified NK cell lines

Increased suitability for standardization

The NK activity may interfere with the measurement of the ADCC-related lysis

Absence of functionally qualified batches of cells

Absence of biosafety qualified batches of cells

To be handled as a GMO

CD16-transduced lymphocytes

Increased suitability for standardization

Qualified batches available in a ready-to-use format

Absence of NK activity-related background

To be handled as a GMO

In this context, the use of standardized effectors should improve significantly the ADCC assays. Previous work has highlighted that human lymphocytes, modified to express the CD16 receptor, have acquired the ADCC functions [1]. Thanks to our specific know-how, clones of CD16+ lymphocytes have been produced on a large scale (109 cellules). The results obtained have pointed out that cells stored in liquid nitrogen and used as soon as they were thawed, were usable for ADCC assays on a reproducible basis (Figure 1). Thanks to that approach, two models of ADCC measurement are characterized in the present presentation: CD20 and Her2neu.
Figure 1

Characterization of ADCC function of CD16-transduced lymphocytes. Two models of antigen expressing cells were used to establish the inter-assay precision of the ADCC measurement in a standard chromium-release assay. Briefly, the cells were 51Cr-labelled, washed, incubated with the antibodies and the release of 51Cr in the supernatant was analyzed after 4h.

The produced effector cells constitute a relevant alternative to replace the use of NK cells when the standardization of ADCC potency assay is needed.

Authors’ Affiliations

Clean Cells


  1. Clémenceau B, Congy-Jolivet N, Gallot G, Vivien R, Gaschet J, Thibault G, Vié H: Antibody-dependent cellular cytotoxicity (ADCC) is mediated by genetically modified antigen-specific human T lymphocytes. Blood. 2006, 107 (12): 4669-77. 10.1182/blood-2005-09-3775.View ArticlePubMedGoogle Scholar


© Bretaudeau and Bonnaudet; licensee BioMed Central Ltd. 2011

This article is published under license to BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.