Details
| Original language | English |
|---|---|
| Article number | e202400057 |
| Number of pages | 12 |
| Journal | Engineering in life sciences |
| Volume | 25 |
| Issue number | 1 |
| Publication status | Published - 9 Jan 2025 |
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.
Keywords
- centrifugal ultrafilters, crossflow cassettes, downstream processing, lentiviral vectors, stirred cells, tangential flow filtration, ultrafiltration
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biotechnology
- Chemical Engineering(all)
- Bioengineering
- Environmental Science(all)
- Environmental Engineering
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In: Engineering in life sciences, Vol. 25, No. 1, e202400057, 09.01.2025.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Investigating Ultrafiltration Membranes and Operation Modes for Improved Lentiviral Vector Processing
AU - Labisch, Jennifer J.
AU - Evangelopoulou, Maria
AU - Schleuß, Tobias
AU - Pickl, Andreas
N1 - Publisher Copyright: © 2024 The Author(s). Engineering in Life Sciences published by Wiley-VCH GmbH.
PY - 2025/1/9
Y1 - 2025/1/9
N2 - 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.
AB - 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.
KW - centrifugal ultrafilters
KW - crossflow cassettes
KW - downstream processing
KW - lentiviral vectors
KW - stirred cells
KW - tangential flow filtration
KW - ultrafiltration
UR - http://www.scopus.com/inward/record.url?scp=85214108072&partnerID=8YFLogxK
U2 - 10.1002/elsc.202400057
DO - 10.1002/elsc.202400057
M3 - Article
AN - SCOPUS:85214108072
VL - 25
JO - Engineering in life sciences
JF - Engineering in life sciences
SN - 1618-0240
IS - 1
M1 - e202400057
ER -