| Autor: |
Leila Mekkaoui, Jose G. Tejerizo, Sara Abreu, Lydie Rubat, Aleksandra Nikoniuk, William Macmorland, Claire Horlock, Sofia Matsumoto, Sarah Williams, Koval Smith, Juliet Price, Saket Srivastava, Rehan Hussain, Mohammad Amin Banani, William Day, Elena Stevenson, Meghan Madigan, Jie Chen, Ravin Khinder, Shahed Miah, Simon Walker, Michael Ade-Onojobi, Sabine Domining, James Sillibourne, Marianna Sabatino, Vladimir Slepushkin, Farzin Farzaneh, Martin Pule |
| Jazyk: |
angličtina |
| Rok vydání: |
2023 |
| Předmět: |
|
| Zdroj: |
Molecular Therapy: Methods & Clinical Development, Vol 28, Iss , Pp 116-128 (2023) |
| Druh dokumentu: |
article |
| ISSN: |
2329-0501 |
| DOI: |
10.1016/j.omtm.2022.12.006 |
| Popis: |
γ-Retroviral vectors (γ-RV) are powerful tools for gene therapy applications. Current clinical vectors are produced from stable producer cell lines which require minimal further downstream processing, while purification schemes for γ-RV produced by transient transfection have not been thoroughly investigated. We aimed to develop a method to purify transiently produced γ-RV for early clinical studies. Here, we report a simple one-step purification method by high-speed centrifugation for γ-RV produced by transient transfection for clinical application. High-speed centrifugation enabled the concentration of viral titers in the range of 107–108 TU/mL with >80% overall recovery. Analysis of research-grade concentrated vector revealed sufficient reduction in product- and process-related impurities. Furthermore, product characterization of clinical-grade γ-RV by BioReliance demonstrated two-logs lower impurities per transducing unit compared with regulatory authority-approved stable producer cell line vector for clinical application. In terms of CAR T cell manufacturing, clinical-grade γ-RV produced by transient transfection and purified by high-speed centrifugation was similar to γ-RV produced from a clinical-grade stable producer cell line. This method will be of value for studies using γ-RV to bridge vector supply between early- and late-stage clinical trials. |
| Databáze: |
Directory of Open Access Journals |
| Externí odkaz: |
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