Harrer, Dennis Christoph and Li, Sin-Syue and Kaljanac, Marcell and Bezler, Valerie and Barden, Markus and Pan, Hong and Herr, Wolfgang and Abken, Hinrich (2024) Magnetic CAR T cell purification using an anti-G4S linker antibody. JOURNAL OF IMMUNOLOGICAL METHODS, 528: 113667. ISSN 0022-1759, 1872-7905
Full text not available from this repository. (Request a copy)Abstract
Chimeric antigen receptor (CAR) redirected T cells are successfully employed in the combat against several hematological malignancies, however, are often compromised by low transduction rates making refinement of the CAR T cell products necessary. Here, we report a broadly applicable enrichment protocol relying on marking CAR T cells with an anti-glycine4-serine (G4S) linker antibody followed by magnetic activated cell sorting (MACS). The protocol is broadly applicable since the G4S peptide is an integral part of the vast majority of CARs as it links the VH and VL recognition domains. We demonstrate the feasibility by using the canonical second generation CARs specific for CEA and Her2, respectively, obtaining highly purified CAR T cell products in a onestep procedure without impairing cell viability. The protocol is also applicable to a dual specific CAR (tandem CAR). Except for CD39, T cell activation/exhaustion markers were not upregulated after separation. Purified CAR T cells retained their functionality with respect to antigen -specific cytokine secretion, cytotoxicity, and the capacity to proliferate and eliminate cognate tumor cells upon repetitive stimulation. Collectively, the one-step protocol for purifying CAR T cells extends the toolbox for preclinical research and specifically for clinical CAR T cell manufacturing.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | RECEPTOR; IMMUNORECEPTORS; ACTIVATION; CAR; Magnetic separation; Isolation; Purification; MACS |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Innere Medizin III (Hämatologie und Internistische Onkologie) |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 28 Jul 2025 11:23 |
| Last Modified: | 28 Jul 2025 11:23 |
| URI: | https://pred.uni-regensburg.de/id/eprint/63856 |
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