Abstract
The purification of virus particles and viral vectors for vaccine and gene therapy applications is gaining increasing importance in order to deliver a fast, efficient, and reliable production process. Ultrafiltration (UF) is a widely employed unit operation in bioprocessing and its use is present in several steps of the downstream purification train of biopharmaceuticals. However, to date few studies have thoroughly investigated the performance of several membrane materials and cut-offs for virus concentration/diafiltration. The present study aimed at developing a novel class of UF cassettes for virus concentration/diafiltration. A detailed study was conducted to evaluate the effects of (i) membrane materials, namely polyethersulfone (PES), regenerated cellulose (RC), and highly cross-linked RC (xRC), (ii) nominal cut-off, and (iii) UF device geometry at different production scales. The results indicate that the xRC cassettes with a cut-off of approximately 500 kDa are able to achieve a 10-fold concentration factor with 100% recovery of particles with a process time twice as fast as that of a commercially available hollow fiber. DNA and host cell protein clearances, as well as hydraulic permeability and fouling behavior, were also assessed.
| Original language | English |
|---|---|
| Article number | e115802 |
| Journal | PlosOne |
| Volume | 9 |
| Issue number | 12 |
| DOIs | |
| Publication status | Published - 29 Dec 2014 |
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Evaluation of novel large cut-off ultrafiltration membranes for adenovirus serotype 5 (Ad5) concentration. / Nestola, Piergiuseppe; Martins, Duarte L.; Peixoto, Cristina; Roederstein, Susanne; Schleuss, Tobias; Alves, Paula M.; Mota, José P B; Carrondo, Manuel J T.
In: PlosOne, Vol. 9, No. 12, e115802, 29.12.2014.Research output: Contribution to journal › Article
TY - JOUR
T1 - Evaluation of novel large cut-off ultrafiltration membranes for adenovirus serotype 5 (Ad5) concentration
AU - Nestola, Piergiuseppe
AU - Martins, Duarte L.
AU - Peixoto, Cristina
AU - Roederstein, Susanne
AU - Schleuss, Tobias
AU - Alves, Paula M.
AU - Mota, José P B
AU - Carrondo, Manuel J T
N1 - WOS:000347120200071
PY - 2014/12/29
Y1 - 2014/12/29
N2 - The purification of virus particles and viral vectors for vaccine and gene therapy applications is gaining increasing importance in order to deliver a fast, efficient, and reliable production process. Ultrafiltration (UF) is a widely employed unit operation in bioprocessing and its use is present in several steps of the downstream purification train of biopharmaceuticals. However, to date few studies have thoroughly investigated the performance of several membrane materials and cut-offs for virus concentration/diafiltration. The present study aimed at developing a novel class of UF cassettes for virus concentration/diafiltration. A detailed study was conducted to evaluate the effects of (i) membrane materials, namely polyethersulfone (PES), regenerated cellulose (RC), and highly cross-linked RC (xRC), (ii) nominal cut-off, and (iii) UF device geometry at different production scales. The results indicate that the xRC cassettes with a cut-off of approximately 500 kDa are able to achieve a 10-fold concentration factor with 100% recovery of particles with a process time twice as fast as that of a commercially available hollow fiber. DNA and host cell protein clearances, as well as hydraulic permeability and fouling behavior, were also assessed.
AB - The purification of virus particles and viral vectors for vaccine and gene therapy applications is gaining increasing importance in order to deliver a fast, efficient, and reliable production process. Ultrafiltration (UF) is a widely employed unit operation in bioprocessing and its use is present in several steps of the downstream purification train of biopharmaceuticals. However, to date few studies have thoroughly investigated the performance of several membrane materials and cut-offs for virus concentration/diafiltration. The present study aimed at developing a novel class of UF cassettes for virus concentration/diafiltration. A detailed study was conducted to evaluate the effects of (i) membrane materials, namely polyethersulfone (PES), regenerated cellulose (RC), and highly cross-linked RC (xRC), (ii) nominal cut-off, and (iii) UF device geometry at different production scales. The results indicate that the xRC cassettes with a cut-off of approximately 500 kDa are able to achieve a 10-fold concentration factor with 100% recovery of particles with a process time twice as fast as that of a commercially available hollow fiber. DNA and host cell protein clearances, as well as hydraulic permeability and fouling behavior, were also assessed.
UR - http://www.scopus.com/inward/record.url?scp=84920070336&partnerID=8YFLogxK
U2 - 10.1371/journal.pone.0115802
DO - 10.1371/journal.pone.0115802
M3 - Article
VL - 9
JO - PlosOne
JF - PlosOne
SN - 1932-6203
IS - 12
M1 - e115802
ER -