Application note: A high-throughput, radioactivity-free assay for cell-mediated cytotoxicity
In this application note, IntelliCyt Corporation demonstrate a fast, efficient, radioactivity-free CMC assay with low sample input requirements, enabling miniaturisation to 384-well plates, using the iQue® Screener…
Immunotherapy promises to be a powerful approach for treating a variety of diseases—most notably cancer.1–5However, development of novel immunotherapeutics has been limited by the lack of high-throughput methods to screen for effective molecular entities or, in the case of adoptive cell therapy, genetically engineered cells. Most current assays that measure cell-mediated cytotoxicity (CMC), such as the chromium release assay, are difficult to perform on large numbers of samples, can only report on a single biological readout like cell membrane integrity, and cannot differentiate between effector and target cells. Methods based on flow cytometry, such as the CFSE assay, 6 can assess CMC at the level of an individual cell and enable discrimination between effector cells and target cells. Traditional flow cytometry is slow, however, making it unsuitable for use as a high throughput screening (HTS) assay.
Here we demonstrate a fast, efficient, radioactivity-free CMC assay with low sample input requirements, enabling miniaturisation to 384-well plates. Using the iQue® Screener and differential labeling of effector and target cells, this method can capture multiple facets of biology—apoptosis markers, signal transduction markers, cell permeability, proliferative capability, and more—in a single well, providing a rich and highly quantitative data set specific to each cell type present. With this approach, researchers and drug discovery teams can quickly screen through compounds and conditions, building a detailed understanding of the molecular events occurring in each well and speeding insight into development of immunotherapeutic approaches.
This application note is restricted - login or subscribe free to access
Thank you for visiting our website. To access this content in full you'll need to login. It's completely free to subscribe, and in less than a minute you can continue reading. If you've already subscribed, great - just login.
Why subscribe? Join our growing community of thousands of industry professionals and gain access to:
- quarterly issues in print and/or digital format
- case studies, whitepapers, webinars and industry-leading content
- breaking news and features
- our extensive online archive of thousands of articles and years of past issues
- ...And it's all free!
Related content from this organisation
- High throughput flow cytometry for multifactorial evaluation of novel therapeutic candidates using an innovative in vitro immunooncology screening platform
- High throughput flow cytometry for multifactorial evaluation of novel therapeutic candidates using an innovative In vitro immuno-oncology screening platform
- Expert view: Addressing the big challenges in drug screening
- Screening In-Depth Focus 2017
- Whitepaper: Advancing therapeutic antibody discovery with multiplexed screening