Automated Large-Scale T Cell Isolation in a New Closed Cell Separation System

Abstract

Abstract Large-scale T cell isolation is commonly performed in a wide range of laboratory settings, including for cell therapy and drug discovery research, as well as in core facilities as part of cell manufacturing and banking. However, current methods can be a significant bottleneck in a lab’s workflow, often requiring a full day just for sample processing and cell isolation. Furthermore, many applications have stringent requirements for purity, sterility, and standardization of the cell isolation procedure. To address these needs, we have developed RoboSep™-C, an instrument for efficient and automated cell isolation in a closed system. RoboSep™-C automates the established EasySep™ technology for immunomagnetic cell separation to enable isolation of untouched T cells, CD4+ T cells, or CD8+ T cells from leukapheresis samples. To set up the system, the user follows the on-screen prompts to install a sterile single-use tubing set and load the recommended medium, cell isolation reagents, and starting leukopak. The instrument then performs all necessary cell processing steps including sample washing, cell labeling, magnetic separation, and cell concentration in a 50-minute protocol. Starting with samples ranging from 2.5 to 20 billion nucleated cells, we obtained purities of 95.4 ± 4.1%, 95.7 ± 2.5%, and 89.3 ± 3.5% for T cell (n=16), CD4+ T cell (n=14), and CD8+ T cell (n=12) isolation, respectively (mean ± SD). The recoveries of the isolated T, CD4+ T, and CD8+ T cells were 64.2 ± 12.4%, 67.0 ± 8.2%, and 54.3 ± 13.5%, respectively. Automated isolation of high-purity T cells with RoboSep™-C enables researchers to scale up their operations and can be easily integrated upstream of existing T cell expansion, genome editing, and cryopreservation protocols.