Details
| Original language | English |
|---|---|
| Article number | 149 |
| Journal | Membranes |
| Volume | 13 |
| Issue number | 2 |
| Publication status | Published - 24 Jan 2023 |
Abstract
Lentiviral vectors (LVs) are widely used in clinical trials of gene and cell therapy. Low LV stability incentivizes constant development and the improvement of gentle process steps. Steric exclusion chromatography (SXC) has gained interest in the field of virus purification but scaling up has not yet been addressed. In this study, the scaling up of lentiviral vector purification by SXC with membrane modules was approached. Visualization of the LVs captured on the membrane during SXC showed predominant usage of the upper membrane layer. Furthermore, testing of different housing geometries showed a strong influence on the uniform usage of the membrane. The main use of the first membrane layer places a completely new requirement on the scaling of the process and the membrane modules. When transferring the SXC process to smaller or larger membrane modules, it became apparent that scaling of the flow rate is a critical factor that must be related to the membrane area of the first layer. Performing SXC at different scales demonstrated that a certain critical minimum surface area-dependent flow rate is necessary to achieve reproducible LV recoveries. With the presented scaling approach, we were able to purify 980 mL LVs with a recovery of 68%.
Keywords
- depletion potential, lentiviral vector purification, membrane chromatography, polyethylene glycol, scaling up of membrane modules, steric exclusion chromatography
ASJC Scopus subject areas
- Chemical Engineering(all)
- Chemical Engineering (miscellaneous)
- Chemical Engineering(all)
- Process Chemistry and Technology
- Chemical Engineering(all)
- Filtration and Separation
Cite this
- Standard
- Harvard
- Apa
- Vancouver
- BibTeX
- RIS
In: Membranes, Vol. 13, No. 2, 149, 24.01.2023.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Scaling Up of Steric Exclusion Membrane Chromatography for Lentiviral Vector Purification
AU - Labisch, Jennifer Julia
AU - Paul, Richard
AU - Wiese, G. Philip
AU - Pflanz, Karl
N1 - Funding Information: This research was funded by Sartorius Stedim Biotech GmbH.
PY - 2023/1/24
Y1 - 2023/1/24
N2 - Lentiviral vectors (LVs) are widely used in clinical trials of gene and cell therapy. Low LV stability incentivizes constant development and the improvement of gentle process steps. Steric exclusion chromatography (SXC) has gained interest in the field of virus purification but scaling up has not yet been addressed. In this study, the scaling up of lentiviral vector purification by SXC with membrane modules was approached. Visualization of the LVs captured on the membrane during SXC showed predominant usage of the upper membrane layer. Furthermore, testing of different housing geometries showed a strong influence on the uniform usage of the membrane. The main use of the first membrane layer places a completely new requirement on the scaling of the process and the membrane modules. When transferring the SXC process to smaller or larger membrane modules, it became apparent that scaling of the flow rate is a critical factor that must be related to the membrane area of the first layer. Performing SXC at different scales demonstrated that a certain critical minimum surface area-dependent flow rate is necessary to achieve reproducible LV recoveries. With the presented scaling approach, we were able to purify 980 mL LVs with a recovery of 68%.
AB - Lentiviral vectors (LVs) are widely used in clinical trials of gene and cell therapy. Low LV stability incentivizes constant development and the improvement of gentle process steps. Steric exclusion chromatography (SXC) has gained interest in the field of virus purification but scaling up has not yet been addressed. In this study, the scaling up of lentiviral vector purification by SXC with membrane modules was approached. Visualization of the LVs captured on the membrane during SXC showed predominant usage of the upper membrane layer. Furthermore, testing of different housing geometries showed a strong influence on the uniform usage of the membrane. The main use of the first membrane layer places a completely new requirement on the scaling of the process and the membrane modules. When transferring the SXC process to smaller or larger membrane modules, it became apparent that scaling of the flow rate is a critical factor that must be related to the membrane area of the first layer. Performing SXC at different scales demonstrated that a certain critical minimum surface area-dependent flow rate is necessary to achieve reproducible LV recoveries. With the presented scaling approach, we were able to purify 980 mL LVs with a recovery of 68%.
KW - depletion potential
KW - lentiviral vector purification
KW - membrane chromatography
KW - polyethylene glycol
KW - scaling up of membrane modules
KW - steric exclusion chromatography
UR - http://www.scopus.com/inward/record.url?scp=85149113862&partnerID=8YFLogxK
U2 - 10.3390/membranes13020149
DO - 10.3390/membranes13020149
M3 - Article
AN - SCOPUS:85149113862
VL - 13
JO - Membranes
JF - Membranes
IS - 2
M1 - 149
ER -