| Autor: |
St-Denis-Bissonnette F; Biologic and Radiopharmaceutical Drugs Directorate; Department of Biochemistry, Microbiology and Immunology Institute, University of Ottawa., Kirkby M; Biologic and Radiopharmaceutical Drugs Directorate; Department of Biochemistry, Microbiology and Immunology Institute, University of Ottawa., Wang L; Department of Biochemistry, Microbiology and Immunology Institute, University of Ottawa; Centre for Infection, Immunity and Inflammation Institute, University of Ottawa; Regenerative Medicine Program, Ottawa Hospital Research Institute; lisheng.wang@uottawa.ca., Lavoie JR; Biologic and Radiopharmaceutical Drugs Directorate; Department of Biochemistry, Microbiology and Immunology Institute, University of Ottawa; jessie.lavoie@hc-sc.gc.ca. |
| Abstrakt: |
Natural killer cell-derived extracellular vesicles (NK-EVs) are being investigated as cancer biotherapeutics. They possess unique properties as cytotoxic nanovesicles targeting cancer cells and as immunomodulatory communicators. A scalable biomanufacturing workflow enables the production of large quantities of high-purity NK-EVs to meet the pre-clinical and clinical demands. The workflow employs a closed-loop hollow-fiber bioreactor, enabling continuous production of NK-EVs from the NK92-MI cell line under serum-free, xeno-free, feeder-free, and antibiotic-free conditions in compliance with Good Manufacturing Practices standards. This protocol-driven study outlines the biomanufacturing workflow for isolating NK-EVs using size-exclusion chromatography, ultrafiltration, and filter-based sterilization. Essential NK-EV product characterization is performed via nanoparticle tracking analysis, and their functionality is assessed through a validated cell viability-based potency assay against cancer cells. This scalable biomanufacturing process holds significant potential to advance the clinical translation of NK-EV-based cancer biotherapeutics by adhering to best practices and ensuring reproducibility. |
