Mekkaoui, Leila;
Tejerizo, Jose G;
Abreu, Sara;
Rubat, Lydie;
Nikoniuk, Aleksandra;
Macmorland, William;
Horlock, Claire;
... Pule, Martin; + view all
(2023)
Efficient clinical-grade γ-retroviral vector purification by high-speed centrifugation for CAR T cell manufacturing.
Molecular Therapy - Methods and Clinical Development
, 28
pp. 116-128.
10.1016/j.omtm.2022.12.006.
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Abstract
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.
| Type: | Article |
|---|---|
| Title: | Efficient clinical-grade γ-retroviral vector purification by high-speed centrifugation for CAR T cell manufacturing |
| Location: | United States |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1016/j.omtm.2022.12.006 |
| Publisher version: | https://doi.org/10.1016/j.omtm.2022.12.006 |
| Language: | English |
| Additional information: | © 2022 The Authors. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
| Keywords: | CAR T cells, downstream processing, high-speed centrifugation, manufacturing, γ-retroviral vectors |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Cancer Institute UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Cancer Institute > Research Department of Haematology |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10163093 |
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