Investigating Ultrafiltration Membranes and Operation Modes for Improved Lentiviral Vector Processing
ABSTRACT The demand for lentiviral vectors (LVs) as tools for ex vivo gene therapies is ever‐increasing. Despite their promising applications, challenges in LV production remain largely due to the fragile envelope, which challenges the maintenance of vector stability. Thus, downstream processing opt...
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| Format: | Article |
| Language: | English |
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Wiley-VCH
2025-01-01
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| Series: | Engineering in Life Sciences |
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| Online Access: | https://doi.org/10.1002/elsc.202400057 |
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| author | Jennifer J. Labisch Maria Evangelopoulou Tobias Schleuß Andreas Pickl |
| author_facet | Jennifer J. Labisch Maria Evangelopoulou Tobias Schleuß Andreas Pickl |
| author_sort | Jennifer J. Labisch |
| collection | DOAJ |
| description | ABSTRACT The demand for lentiviral vectors (LVs) as tools for ex vivo gene therapies is ever‐increasing. Despite their promising applications, challenges in LV production remain largely due to the fragile envelope, which challenges the maintenance of vector stability. Thus, downstream processing optimization to enhance efficiency, yield, and product quality is necessary. This study investigated the influence of membrane types and filtration devices during ultrafiltration (UF). Nine different membrane materials consisting of polyethersulfone (PES), regenerated cellulose, or Hydrosart, with distinct molecular weight cutoffs, were evaluated in stirred cells, centrifugal ultrafilters, and crossflow cassettes. The evaluation was based on the ability to retain infectious LV particles and remove impurities. The analysis revealed that a reinforced 100 kDa PES and a 300 kDa Hydrosart membrane had the best overall ability to concentrate infectious LVs and remove DNA, especially when operated in a stirred cell. Challenges were seen in the nonoptimized crossflow cassette process, where infectious LV recovery was generally lower compared to other devices. We demonstrated that membrane material and filtration device have a direct impact on the efficiency of LV UF. |
| format | Article |
| id | doaj-art-63c6debeaf174c4b800069a76f95e13c |
| institution | Kabale University |
| issn | 1618-0240 1618-2863 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Wiley-VCH |
| record_format | Article |
| series | Engineering in Life Sciences |
| spelling | doaj-art-63c6debeaf174c4b800069a76f95e13c2025-01-30T06:40:30ZengWiley-VCHEngineering in Life Sciences1618-02401618-28632025-01-01251n/an/a10.1002/elsc.202400057Investigating Ultrafiltration Membranes and Operation Modes for Improved Lentiviral Vector ProcessingJennifer J. Labisch0Maria Evangelopoulou1Tobias Schleuß2Andreas Pickl3Lab Essentials Applications Development Sartorius Göttingen GermanyLab Essentials Applications Development Sartorius Göttingen GermanyUltrafiltration Membrane Technology Sartorius Göttingen GermanyLab Essentials Applications Development Sartorius Göttingen GermanyABSTRACT The demand for lentiviral vectors (LVs) as tools for ex vivo gene therapies is ever‐increasing. Despite their promising applications, challenges in LV production remain largely due to the fragile envelope, which challenges the maintenance of vector stability. Thus, downstream processing optimization to enhance efficiency, yield, and product quality is necessary. This study investigated the influence of membrane types and filtration devices during ultrafiltration (UF). Nine different membrane materials consisting of polyethersulfone (PES), regenerated cellulose, or Hydrosart, with distinct molecular weight cutoffs, were evaluated in stirred cells, centrifugal ultrafilters, and crossflow cassettes. The evaluation was based on the ability to retain infectious LV particles and remove impurities. The analysis revealed that a reinforced 100 kDa PES and a 300 kDa Hydrosart membrane had the best overall ability to concentrate infectious LVs and remove DNA, especially when operated in a stirred cell. Challenges were seen in the nonoptimized crossflow cassette process, where infectious LV recovery was generally lower compared to other devices. We demonstrated that membrane material and filtration device have a direct impact on the efficiency of LV UF.https://doi.org/10.1002/elsc.202400057centrifugal ultrafilterscrossflow cassettesdownstream processinglentiviral vectorsstirred cellstangential flow filtration |
| spellingShingle | Jennifer J. Labisch Maria Evangelopoulou Tobias Schleuß Andreas Pickl Investigating Ultrafiltration Membranes and Operation Modes for Improved Lentiviral Vector Processing Engineering in Life Sciences centrifugal ultrafilters crossflow cassettes downstream processing lentiviral vectors stirred cells tangential flow filtration |
| title | Investigating Ultrafiltration Membranes and Operation Modes for Improved Lentiviral Vector Processing |
| title_full | Investigating Ultrafiltration Membranes and Operation Modes for Improved Lentiviral Vector Processing |
| title_fullStr | Investigating Ultrafiltration Membranes and Operation Modes for Improved Lentiviral Vector Processing |
| title_full_unstemmed | Investigating Ultrafiltration Membranes and Operation Modes for Improved Lentiviral Vector Processing |
| title_short | Investigating Ultrafiltration Membranes and Operation Modes for Improved Lentiviral Vector Processing |
| title_sort | investigating ultrafiltration membranes and operation modes for improved lentiviral vector processing |
| topic | centrifugal ultrafilters crossflow cassettes downstream processing lentiviral vectors stirred cells tangential flow filtration |
| url | https://doi.org/10.1002/elsc.202400057 |
| work_keys_str_mv | AT jenniferjlabisch investigatingultrafiltrationmembranesandoperationmodesforimprovedlentiviralvectorprocessing AT mariaevangelopoulou investigatingultrafiltrationmembranesandoperationmodesforimprovedlentiviralvectorprocessing AT tobiasschleuß investigatingultrafiltrationmembranesandoperationmodesforimprovedlentiviralvectorprocessing AT andreaspickl investigatingultrafiltrationmembranesandoperationmodesforimprovedlentiviralvectorprocessing |