Continuous manufacturing of lentiviral vectors using a stable producer cell line in a fixed-bed bioreactor
Continuous manufacturing of lentiviral vectors (LVs) using stable producer cell lines could extend production periods, improve batch-to-batch reproducibility, and eliminate costly plasmid DNA and transfection reagents. A continuous process was established by expanding cells constitutively expressing...
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| Format: | Article |
| Language: | English |
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Elsevier
2024-03-01
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| Series: | Molecular Therapy: Methods & Clinical Development |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2329050124000251 |
| _version_ | 1797294279895285760 |
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| author | Dale J. Stibbs Pedro Silva Couto Yasuhiro Takeuchi Qasim A. Rafiq Nigel B. Jackson Andrea C.M.E. Rayat |
| author_facet | Dale J. Stibbs Pedro Silva Couto Yasuhiro Takeuchi Qasim A. Rafiq Nigel B. Jackson Andrea C.M.E. Rayat |
| author_sort | Dale J. Stibbs |
| collection | DOAJ |
| description | Continuous manufacturing of lentiviral vectors (LVs) using stable producer cell lines could extend production periods, improve batch-to-batch reproducibility, and eliminate costly plasmid DNA and transfection reagents. A continuous process was established by expanding cells constitutively expressing third-generation LVs in the iCELLis Nano fixed-bed bioreactor. Fixed-bed bioreactors provide scalable expansion of adherent cells and enable a straightforward transition from traditional surface-based culture vessels. At 0.5 vessel volume per day (VVD), the short half-life of LVs resulted in a low total infectious titer at 1.36 × 104 TU cm−2. Higher perfusion rates increased titers, peaking at 7.87 × 104 TU cm−2 at 1.5 VVD. The supernatant at 0.5 VVD had a physical-to-infectious particle ratio of 659, whereas this was 166 ± 15 at 1, 1.5, and 2 VVD. Reducing the pH from 7.20 to 6.85 at 1.5 VVD improved the total infectious yield to 9.10 × 104 TU cm−2. Three independent runs at 1.5 VVD and a culture pH of 6.85 showed low batch-to-batch variability, with a coefficient of variation of 6.4% and 10.0% for total infectious and physical LV yield, respectively. This study demonstrated the manufacture of high-quality LV supernatant using a stable producer cell line that does not require induction. |
| first_indexed | 2024-03-07T21:28:53Z |
| format | Article |
| id | doaj.art-9a07beca55a94568a5587afb1f9acd79 |
| institution | Directory Open Access Journal |
| issn | 2329-0501 |
| language | English |
| last_indexed | 2024-03-07T21:28:53Z |
| publishDate | 2024-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Methods & Clinical Development |
| spelling | doaj.art-9a07beca55a94568a5587afb1f9acd792024-02-27T04:19:50ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012024-03-01321101209Continuous manufacturing of lentiviral vectors using a stable producer cell line in a fixed-bed bioreactorDale J. Stibbs0Pedro Silva Couto1Yasuhiro Takeuchi2Qasim A. Rafiq3Nigel B. Jackson4Andrea C.M.E. Rayat5Department of Biochemical Engineering, University College London, Bernard Katz Building, Gower Street, London WC1E 6BT, UKDepartment of Biochemical Engineering, University College London, Bernard Katz Building, Gower Street, London WC1E 6BT, UKDivision of Infection and Immunity, University College London, Cruciform Building, Gower Street, London WC1E 6BT, UK; Biotherapeutics and Advanced Therapies, Scientific Research and Innovation, Medicines and Healthcare Products Regulatory Agency, South Mimms, Potters Bar EN6 3QC, UKDepartment of Biochemical Engineering, University College London, Bernard Katz Building, Gower Street, London WC1E 6BT, UKCytiva, 5 Harbourgate Business Park, Southampton Road, Portsmouth PO6 4BQ, UKDepartment of Biochemical Engineering, University College London, Bernard Katz Building, Gower Street, London WC1E 6BT, UK; Corresponding author: Andrea C.M.E. Rayat, Department of Biochemical Engineering, University College London, Bernard Katz Building, Gower Street, London WC1E 6BT, UK.Continuous manufacturing of lentiviral vectors (LVs) using stable producer cell lines could extend production periods, improve batch-to-batch reproducibility, and eliminate costly plasmid DNA and transfection reagents. A continuous process was established by expanding cells constitutively expressing third-generation LVs in the iCELLis Nano fixed-bed bioreactor. Fixed-bed bioreactors provide scalable expansion of adherent cells and enable a straightforward transition from traditional surface-based culture vessels. At 0.5 vessel volume per day (VVD), the short half-life of LVs resulted in a low total infectious titer at 1.36 × 104 TU cm−2. Higher perfusion rates increased titers, peaking at 7.87 × 104 TU cm−2 at 1.5 VVD. The supernatant at 0.5 VVD had a physical-to-infectious particle ratio of 659, whereas this was 166 ± 15 at 1, 1.5, and 2 VVD. Reducing the pH from 7.20 to 6.85 at 1.5 VVD improved the total infectious yield to 9.10 × 104 TU cm−2. Three independent runs at 1.5 VVD and a culture pH of 6.85 showed low batch-to-batch variability, with a coefficient of variation of 6.4% and 10.0% for total infectious and physical LV yield, respectively. This study demonstrated the manufacture of high-quality LV supernatant using a stable producer cell line that does not require induction.http://www.sciencedirect.com/science/article/pii/S2329050124000251lentiviral vectorstable producer cell linecontinuous bioprocessingfixed-bed bioreactorperfusion culture |
| spellingShingle | Dale J. Stibbs Pedro Silva Couto Yasuhiro Takeuchi Qasim A. Rafiq Nigel B. Jackson Andrea C.M.E. Rayat Continuous manufacturing of lentiviral vectors using a stable producer cell line in a fixed-bed bioreactor Molecular Therapy: Methods & Clinical Development lentiviral vector stable producer cell line continuous bioprocessing fixed-bed bioreactor perfusion culture |
| title | Continuous manufacturing of lentiviral vectors using a stable producer cell line in a fixed-bed bioreactor |
| title_full | Continuous manufacturing of lentiviral vectors using a stable producer cell line in a fixed-bed bioreactor |
| title_fullStr | Continuous manufacturing of lentiviral vectors using a stable producer cell line in a fixed-bed bioreactor |
| title_full_unstemmed | Continuous manufacturing of lentiviral vectors using a stable producer cell line in a fixed-bed bioreactor |
| title_short | Continuous manufacturing of lentiviral vectors using a stable producer cell line in a fixed-bed bioreactor |
| title_sort | continuous manufacturing of lentiviral vectors using a stable producer cell line in a fixed bed bioreactor |
| topic | lentiviral vector stable producer cell line continuous bioprocessing fixed-bed bioreactor perfusion culture |
| url | http://www.sciencedirect.com/science/article/pii/S2329050124000251 |
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