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Linker-specific monoclonal antibodies present a simple and reliable detection method for scFv-based CARNK cells
Linker-specific monoclonal antibodies present a simple and reliable detection method for scFv-based CARNK cells
Katharina Schindler
(Department for Cell and Gene Therapy Development, Fraunhofer Institute for Cell Therapy and Immunology (IZI), 04103 Leipzig, Germany)
Katharina Eva Ruppel
(Department for Cell and Gene Therapy Development, Fraunhofer Institute for Cell Therapy and Immunology (IZI), 04103 Leipzig, Germany)
Claudia Müller
(Department of Preclinical Development and Validation, Fraunhofer Institute for Cell Therapy and Immunology (IZI), 04103 Leipzig, Germany; Fraunhofer Cluster of Excellence Immune-Mediated Diseases, 60596 Frankfurt, Germany)
Ulrike Koehl
(Department for Cell and Gene Therapy Development, Fraunhofer Institute for Cell Therapy and Immunology (IZI), 04103 Leipzig, Germany; Institute for Clinical Immunology, University of Leipzig, 04103 Leipzig, Germany; Fraunhofer Cluster of Excellence Immune-Mediated Diseases, 60596 Frankfurt, Germany)
Stephan Fricke
(Department for Cell and Gene Therapy Development, Fraunhofer Institute for Cell Therapy and Immunology (IZI), 04103 Leipzig, Germany; Fraunhofer Cluster of Excellence Immune-Mediated Diseases, 60596 Frankfurt, Germany; Medicine Campus MEDiC of the Technical University of Dresden at Klinikum Chemnitz gGmbH, 09116 Chemnitz, Germany)
Dominik Schmiedel
(Department for Cell and Gene Therapy Development, Fraunhofer Institute for Cell Therapy and Immunology (IZI), 04103 Leipzig, Germany; Institute for Clinical Immunology, University of Leipzig, 04103 Leipzig, Germany; Corresponding author: Dominik Schmiedel, Department for Cell and Gene Therapy Development, Fraunhofer Institute for Cell Therapy and Immunology (IZI), 04103 Leipzig, Germany.)
Chimeric antigen receptor (CAR) T cell therapies have demonstrated significant successes in treating cancer. Currently, there are six approved CAR T cell products available on the market that target different malignancies of the B cell lineage. However, to overcome the limitations of CAR T cell therapies, other immune cells are being investigated for CAR-based cell therapies. CAR natural killer (NK) cells can be applied as allogeneic cell therapy, providing an economical, safe, and efficient alternative to autologous CAR T cells. To improve CAR research and future in-patient monitoring of cell therapeutics, a simple, reliable, and versatile CAR detection reagent is crucial. As most existing CARs contain a single-chain variable fragment (scFv) with either a Whitlow or a G4S linker site, linker-specific monoclonal antibodies (mAbs) can detect a broad range of CARs. This study demonstrates that these linker-specific mAbs can detect different CAR NK cells in vitro, spiked in whole blood, and within patient-derived tumor spheroids with high specificity and sensitivity, providing an effective and almost universal alternative for scFv-based CAR detection. Additionally, we confirm that linker-specific antibodies can be used for functional testing and enrichment of CAR NK cells, thereby providing a useful research tool to fast-track the development of novel CAR-based therapies.