title: Automated Enrichment, Transduction, and Expansion of Clinical-Scale CD62L+T Cells for Manufacturing of Gene Therapy Medicinal Products.
abstract: Multiple clinical studies have demonstrated that adaptive immunotherapy using redirected T cells against advanced cancer has led to promising results with improved patient survival. The continuously increasing interest in those advanced gene therapy medicinal products (GTMPs) leads to a manufacturing challenge regarding automation, process robustness, and cell storage. Therefore, this study addresses the proof of principle in clinical-scale selection, stimulation, transduction, and expansion of T cells using the automated closed CliniMACS®Prodigy system. Naïve and central memory T cells from apheresis products were first immunomagnetically enriched using anti-CD62L magnetic beads and further processed freshly (n = 3) or split for cryopreservation and processed after thawing (n = 1). Starting with 0.5 × 108purified CD3+T cells, three mock runs and one run including transduction with green fluorescent protein (GFP)-containing vector resulted in a median final cell product of 16 × 108T cells (32-fold expansion) up to harvesting after 2 weeks. Expression of CD62L was downregulated on T cells after thawing, which led to the decision to purify CD62L+CD3+T cells freshly with cryopreservation thereafter. Most important in the split product, a very similar expansion curve was reached comparing the overall freshly CD62L selected cells with those after thawing, which could be demonstrated in the T cell subpopulations as well by showing a nearly identical conversion of the CD4/CD8 ratio. In the GFP run, the transduction efficacy was 83%. In-process control also demonstrated sufficient glucose levels during automated feeding and medium removal. The robustness of the process and the constant quality of the final product in a closed and automated system give rise to improve harmonized manufacturing protocols for engineered T cells in future gene therapy studies.
authors: 1 Cellular Therapy Center, Institute of Cellular Therapeutics , Hannover Medical School, Hannover, Germany.
authors: Christoph
authors: C
authors: Priesner
authors: 1 Cellular Therapy Center, Institute of Cellular Therapeutics , Hannover Medical School, Hannover, Germany.
authors: Krasimira
authors: K
authors: Aleksandrova
authors: 2 GMP Development Unit, Institute of Cellular Therapeutics , Integrated Research and Treatment Center for Transplantation, Hannover Medical School, Hannover, Germany.
authors: Ruth
authors: R
authors: Esser
authors: 3 Miltenyi Biotec, Bergisch Gladbach, Germany .
authors: Nadine
authors: N
authors: Mockel-Tenbrinck
authors: 1 Cellular Therapy Center, Institute of Cellular Therapeutics , Hannover Medical School, Hannover, Germany.
authors: Jana
authors: J
authors: Leise
authors: 3 Miltenyi Biotec, Bergisch Gladbach, Germany .
authors: Katharina
authors: K
authors: Drechsel
authors: 2 GMP Development Unit, Institute of Cellular Therapeutics , Integrated Research and Treatment Center for Transplantation, Hannover Medical School, Hannover, Germany.
authors: Michael
authors: M
authors: Marburger
authors: 2 GMP Development Unit, Institute of Cellular Therapeutics , Integrated Research and Treatment Center for Transplantation, Hannover Medical School, Hannover, Germany.
authors: Andrea
authors: A
authors: Quaiser
authors: 4 Institute of Transfusion Medicine , Hannover Medical School, Hannover, Germany.
authors: Lilia
authors: L
authors: Goudeva
authors: 1 Cellular Therapy Center, Institute of Cellular Therapeutics , Hannover Medical School, Hannover, Germany.
authors: Lubomir
authors: L
authors: Arseniev
authors: 3 Miltenyi Biotec, Bergisch Gladbach, Germany .
authors: Andrew D
authors: AD
authors: Kaiser
authors: 2 GMP Development Unit, Institute of Cellular Therapeutics , Integrated Research and Treatment Center for Transplantation, Hannover Medical School, Hannover, Germany.
authors: Wolfgang
authors: W
authors: Glienke
authors: 1 Cellular Therapy Center, Institute of Cellular Therapeutics , Hannover Medical School, Hannover, Germany.|2 GMP Development Unit, Institute of Cellular Therapeutics , Integrated Research and Treatment Center for Transplantation, Hannover Medical School, Hannover, Germany.
authors: Ulrike
authors: U
authors: Koehl
mesh_terms: D015316:Genetic Therapy; D005947:Glucose / Q000378:metabolism; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods; D019041:L-Selectin / Q000096:biosynthesis* / Q000235:genetics / Q000627:therapeutic use; D013601:T-Lymphocytes / Q000378:metabolism* / Q000637:transplantation; D014161:Transduction, Genetic
chemical_list: D019041:L-Selectin; D005947:Glucose