Releasable RapidSpheres™ enable immunomagnetic purification of highly viable and functional immune cells from complex tissues in less than 30 minutes

• Published in: The Journal of immunology (1950) Vol. 196; no. 1_Supplement; pp. 69 - 69.14
• Main Authors: Clarke, Samuel, Kokaji, Andy I, Kellerman, Drew W, Ewen, Catherine, Chambers, Martina N, Chan, Mandy, Woodside, Steven M
• Format: Journal Article
• Language: English
• Published: 01-05-2016
• ISSN: 0022-1767; 1550-6606
• DOI: 10.4049/jimmunol.196.Supp.69.14

Abstract
Recent advances in cellular therapy and diagnostics promise dramatic improvements in disease prevention and treatment. This research is driven by scientists studying how cells of the immune system communicate and relies on the success of cell-based assays. However, it remains challenging and time-consuming to purify large numbers of high-quality cells from complex tissues. EasySep™ Release is a fast and easy cell isolation method which utilizes the Releasable RapidSpheres™. This novel magnetic particle technology improves cell purities by reducing contaminants and allows for gentle particle removal to mitigate potential interference in downstream assays. The 29-minute protocol involves first incubating target cells with antibody complexes followed by the Releasable RapidSpheres™. Next, labelled cells are purified using a hand-held magnet. Particle-free purified cells are obtained by applying a mild dissociation reagent and a final magnetic separation. We validated EasySep™ Release by purifying T, B and NK cells from peripheral blood mononuclear cell samples containing 5 to 800 million cells. Cell purities were 99.7 ± 0.1% (n=17) for CD3+ T cells, 99.4 ± 0.4% (n=17) for CD4+ cells, 98.8 ± 0.5% (n=17) for CD8+ T cells, 98.6 ± 0.7% (n=6) for CD19+ cells and 95.6 ± 1.7% (n=6) for CD56+ cells. Isolated cells were confirmed particle-free, viable and functional. We obtained similar high performance using unprocessed leukapheresis samples of up to 5 billion cells, demonstrating excellent scalability and compatibility with more complex samples. Finally, we show how EasySep™ Release can be paired with commercially available antibodies or sequential separations to isolate almost any cell type, including those with a complex phenotype.