Promega's Cookie Policy

We use cookies and similar technologies to make our website work, run analytics, improve our website, and show you personalized content and advertising. Some of these cookies are essential for our website to work. For others, we won’t set them unless you accept them. To find out more about cookies and how to manage cookies, read our Cookie Policy.

Measurement of Fc-mediated ADCC and CDC of anti-TNFα and anti-VEGF Therapeutic Antibodies using Reporter-based Bioassays and Engineered TNFα+ and VEGF+ Target Cells

Part # PS335


Denise Garvin, Rich Moravec, Jamison Grailer, Jim Hartnett, Chris Heid, Frank Fan, Mei Cong and Zhi-jie Jey Cheng
Promega Corporation, 2800 Woods Hollow Rd, Madison, WI 53711

To measure ADCC activity of anti-TNFα and anti-VEGF blocking antibodies, we developed engineered target cells that express either membrane-bound TNFα or VEGF. When used as target cells with reporter-based effector cells expressing a relevant FcgR, ADCC activity of adalimumab (anti-TNFα) and bevacizumab (anti-VEGF) was detected in a specific and dose-dependent manner. Similarly, when used in a luminescence-based CDC assay, the engineered target cells elicited an appropriate FcgR-mediated response. The assay signals demonstrated IgG isotype specificity as IgG4 variants showed minimal activity in both ADCC and CDC assays. Our results demonstrate that the combined use of cell-based reporter bioassays with target cells engineered to express membrane-bound soluble ligands can provide a simple, specific, and quantitative platform to measure Fc-mediated effector functions of therapeutic antibodies targeting soluble ligands.

Printed in USA.