About Antibody-Dependent Cellular Cytotoxicity (ADCC)<\/h2>\n Antibody-dependent cellular cytotoxicity (ADCC), also referred to as antibody-dependent cell-mediated cytotoxicity, is a cytolytic function by which effector cells with cytotoxic potential, such as Natural Killer cells, lyse target cells.\u00a0 This function is dependent on an antibody, which serves as a bridge between the target cell and the effector cell.\u00a0 Mechanistically, the antibody binds its cognate antigen on the target cell through its antigen recognition domain, and then recruits the effector cell to the target with its Fc region.\u00a0 Once bound, the effector cell is in proximity of the target cell and can secrete cytolytic molecules to kill it.<\/p>\n
ADCC can play roles in general human health and mechanism of action for therapeutic antibodies.\u00a0 \u00a0First, it eliminates foreign pathogens and aberrant cells that would otherwise promote illness in general human health.\u00a0 Second, it is suggested that ADCC is a mechanism of action for many therapeutic antibodies, such as those that target HER2, CD20, and CD52.\u00a0 During drug development, potential therapeutic antibody candidates will be screened for ADCC activity to find ones that will have the most potent anti-cancer abilities.<\/p>\n
iQ\u2019s ADCC Assays:\u00a0 Lab-Tested, Immunologist-Approved<\/h2>\n iQ has extensive experience with ADCC assays and can help you design the best experiment, keeping in mind factors that influence ADCC to determine the ADCC potential of your therapeutic antibodies. \u00a0Because there are multiple components to ADCC, three well-studied factors can enhance or decrease ADCC:<\/p>\n
\nGlycosylation state<\/strong> \u2013 antibodies are glycosylated and the oligosaccharides expressed on them can affect ADCC<\/li>\nFcgRIIIa (CD16) polymorphism<\/strong> \u2013 the FcgRIIIa binds the Fc region of the antibody, which comes in two polymorphisms wherein the hi-affinity valine form binds stronger than the low-affinity phenylalanine form and leads to more potent ADCC<\/li>\nExpression level of antigen on target cells<\/strong> \u2013 the target cell line should express sufficient target antigen for the antibodies to form immune complexes so activation signals can be elicited to drive ADCC.<\/li>\n<\/ol>\nAt iQ, we have moved away from traditional cytotoxicity assays that typically load target cells with radioactive chromium, which is released upon lysis.\u00a0 Instead, we primarily perform ADCC assays by high-throughput flow cytometry, wherein target cells are labeled with a fluorescent dye, to differentiate the targets from effector cells.\u00a0 After the target and effector cell co-culture, samples are analyzed with the flow cytometer along with a viability dye to assess the loss of membrane integrity of target cells in the culture, as a measure of cytotoxicity.\u00a0<\/p>\n
An alternative method that we employ at iQ is to use genetically engineered effector reporter cell lines that express luciferase upon FcgRIIIa binding to antibody.\u00a0 With this method, luciferase expression is measured instead of cell death as these effector cells are non-cytotoxic.\u00a0\u00a0 This method provides the advantage of using a stable, reproducible effector cell line as compared to human effector cells, which can vary in activity from donor to donor.<\/p>\n
With our extensive knowledge of factors that influence ADCC and our wide array of readouts for ADCC, iQ can help design the right experiments for you to determine the ADCC potential of your therapeutic antibodies.<\/p>\n<\/div>
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