UNIT 7.27 51Cr Release Assay of Antibody-Dependent Cell-Mediated Cytotoxicity (ADCC)
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Immunology
How to Cite
Nelson, D. L., Kurman, C. C. and Serbousek, D. E. 2001. 51Cr Release Assay of Antibody-Dependent Cell-Mediated Cytotoxicity (ADCC). Current Protocols in Immunology. 8:IV:7.27:7.27.1–7.27.8.
- Published Online: 1 MAY 2001
- Published Print: DEC 1993
Antibody-dependent cell-mediated cytotoxicity (ADCC) is an immunologic cytotoxic effector mechanism that is dependent on the cooperative interaction of humoral and cellular effector elements. This unit describes an assay of ADCC activity that can be used as a test for immunocompetence in effector cells or to test the activity of a monoclonal antibody to mediate ADCC. In this form of cytotoxicity, effector cells with receptors for the Fc portion of immunoglobulin produce target cell lysis by attachment to the Fc portion of antibodies that are bound to target cells via their antigen-combining sites. Therefore, an ADCC assay involves three essential components: labeled target cells, antibodies with specificity for target-cell surface antigens, and effector-cell populations. The basic protocol describes a method of measuring ADCC effector activity in lymphoid cells (peripheral blood mononuclear cells, or PBMC) that employs 51Cr-labeled target cells. The three components are mixed in microtiter-plate wells and lysis of the target cells is detected by measuring the release of radioactivity into the cell supernatant. Support protocols describe procedures for preparing anti-target cell antiserum and 51Cr-labeled target cells.