ADCC

Promega Fc Effector Assays: Measure Every Mechanism

Posted on by Promega

This post is written by Kai Hillman, PhD, Promega Corporation.

Every day, scientists push the boundaries of what’s possible with monoclonal antibodies (mAbs)—from targeting cancer cells to calming autoimmune-driven inflammation. These therapies rely not only on binding but on engineering the desired immune response. The suite of Promega Fc Effector Assays helps you understand these interactions from receptor binding and function, through bridging studies. With consistency, sensitivity, and scalability, these assays support teams from early discovery through lot release.

This article draws on real-world publications and product insights to show how Promega assays are powering next-generation immunotherapies—and redefining how we measure immune engagement.

Schematic diagramming the suite of Promega Fc effector assays in one seamless workflow to support antibody development across the pipeline.
Figure 1. Promega delivers the most comprehensive suite of Fc effector assays in one seamless workflow to support antibody development across the pipeline.
Continue reading “Promega Fc Effector Assays: Measure Every Mechanism”

How Can a ADCC Reporter Bioassay Help You Save Time and Reduce Variability?

Posted on by Kari Kenefick

Fc receptor-mediated antibody-dependent cell-mediated cytotoxicity (ADCC) is an important mechanism of action (MOA) by which antibodies target diseased cells for elimination. Traditional methods for measuring ADCC require primary donor peripheral blood mononuclear cells (PBMCs) or purified natural killer (NK) cells that express Fc receptors on the cell surface. However, primary cultures of PBMCs and NK cells introduce variability, high background, and can be tedious to prepare. Using a commercially available ADCC reporter bioassay can overcome many of the limitations of these primary cell assays.

Our ADCC and ADCP Reporter Bioassays are biologically relevant, MOA-based assays that can be used to measure the potency and stability of antibodies and other biologics that specifically bind and activate Fcγ receptors. The ADCC Reporter Bioassays use an alternative readout from traditional primary cell-based assays: the FcγR and NFAT-mediated activation of luciferase activity in the effector cells. Primary cells are replaced with a Jurkat cell line stably expressing human FcγR variant and NFAT-induced luciferase.

The thaw-and-use cell format of the ADCC Reporter Bioassay saves time and labor of primary cell assays, while reducing variability. While a primary cell assay can take 1-2 weeks from culturing cells to results, ADCC reporter bioassay can be performed in 3–24 hours. The bioassays include all of the required reagents and are easily amenable to high-throughput workflows, enabling you to have precisely the right throughput for your workflow needs.

Check out the full Promega portfolio of Fc effector reporter bioassays to discover the best tool for your research and read more about how these assays

ADCC Reporter Bioassay Makes Top 10 Innovator List

Posted on by Michele Arduengo, PhD
The ADCC Reporter Bioassays were named a Top 10 innovation by The Scientist Magazine.
The ADCC Reporter Bioassay systems were named a Top 10 innovation by The Scientist Magazine.

For the second year running a Promega technology has made The Scientist Magazine’s list of Top 10 Innovations. In 2012 NanoLuc® luciferase technology was in the spotlight; in 2013 the ADCC Reporter Bioassay took center stage.

Antibody-dependent cell-mediated cytotoxicity (ADCC) is the main mechanism of action (MOA) of antibodies through which virus-infected or other diseased cells are targeted for destruction by components of the cell-mediated immune system. These assays are often used to assess the effectiveness of monoclonal antibody therapies during the manufacture and development of biologic drugs. The bioluminescent assays use an alternative readout at an earlier point in ADCC MOA pathway for the quantification of Fc effector function of antibody-based molecules: the activation of gene transcription through the NFAT (nuclear factor of activated T-cells) pathway in the effector cell.

The bioassay uses engineered Jurkat cells stably expressing the FcγRIIIa receptor, V158 (high affinity) variant, and an NFAT response element driving expression of firefly luciferase. The assay is MOA-based and features frozen, thaw-and-use effector cells and optimized reagents and protocol to perform a reporter-based ADCC bioassay in a single day. The bioassay correlates with classic cytotoxic ADCC assays and is a suitable replacement for these cumbersome and highly variable assays.

The novel bioassay is linear, accurate, precise and stability indicating. Moreover, the bioassay shows good linear correlation between levels of glycosylation or fucosylation and ADCC activity. All of these features indicate the assay is suitable for use across biologic drug development programs.