Selection and optimization of transfection enhancer additives for increased virus-like particle production in HEK293 suspension cell cultures
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| 024 | 7 | 0 | |a 10.1007/s00253-015-6842-4 |2 doi |
| 035 | |a (NATIONALLICENCE)springer-10.1007/s00253-015-6842-4 | ||
| 245 | 0 | 0 | |a Selection and optimization of transfection enhancer additives for increased virus-like particle production in HEK293 suspension cell cultures |h [Elektronische Daten] |c [Laura Cervera, Javier Fuenmayor, Irene González-Domínguez, Sonia Gutiérrez-Granados, Maria Segura, Francesc Gòdia] |
| 520 | 3 | |a The manufacturing of biopharmaceuticals in mammalian cells typically relies on the use of stable producer cell lines. However, in recent years, transient gene expression has emerged as a suitable technology for rapid production of biopharmaceuticals. Transient gene expression is particularly well suited for early developmental phases, where several potential therapeutic targets need to be produced and tested in vivo. As a relatively new bioprocessing modality, a number of opportunities exist for improving cell culture productivity upon transient transfection. For instance, several compounds have shown positive effects on transient gene expression. These transfection enhancers either facilitate entry of PEI/DNA transfection complexes into the cell or nucleus or increase levels of gene expression. In this work, the potential of combining transfection enhancers to increase Gag-based virus-like particle production levels upon transfection of suspension-growing HEK 293 cells is evaluated. Using Plackett-Burman design of experiments, it is first tested the effect of eight transfection enhancers: trichostatin A, valproic acid, sodium butyrate, dimethyl sulfoxide (DMSO), lithium acetate, caffeine, hydroxyurea, and nocodazole. An optimal combination of compounds exhibiting the highest effect on gene expression levels was subsequently identified using a surface response experimental design. The optimal consisted on the addition of 20mM lithium acetate, 3.36mM valproic acid, and 5.04mM caffeine which increased VLP production levels 3.8-fold, while maintaining cell culture viability at 94%. | |
| 540 | |a Springer-Verlag Berlin Heidelberg, 2015 | ||
| 690 | 7 | |a VLPs |2 nationallicence | |
| 690 | 7 | |a Production additives |2 nationallicence | |
| 690 | 7 | |a HEK 293 |2 nationallicence | |
| 690 | 7 | |a Transient transfection |2 nationallicence | |
| 690 | 7 | |a Design of experiments |2 nationallicence | |
| 700 | 1 | |a Cervera |D Laura |u Grup d'Enginyeria Cellular i Bioprocés, Escola d'Enginyeria, Universitat Autònoma de Barcelona, Campus de Bellaterra, Cerdanyola del Vallès, 08193, Barcelona, Spain |4 aut | |
| 700 | 1 | |a Fuenmayor |D Javier |u Grup d'Enginyeria Cellular i Bioprocés, Escola d'Enginyeria, Universitat Autònoma de Barcelona, Campus de Bellaterra, Cerdanyola del Vallès, 08193, Barcelona, Spain |4 aut | |
| 700 | 1 | |a González-Domínguez |D Irene |u Grup d'Enginyeria Cellular i Bioprocés, Escola d'Enginyeria, Universitat Autònoma de Barcelona, Campus de Bellaterra, Cerdanyola del Vallès, 08193, Barcelona, Spain |4 aut | |
| 700 | 1 | |a Gutiérrez-Granados |D Sonia |u Grup d'Enginyeria Cellular i Bioprocés, Escola d'Enginyeria, Universitat Autònoma de Barcelona, Campus de Bellaterra, Cerdanyola del Vallès, 08193, Barcelona, Spain |4 aut | |
| 700 | 1 | |a Segura |D Maria |u Bluebird Bio Pharmaceutical Sciences, Process Development Group, 150 2nd Street, 02141, Cambridge, MA, USA |4 aut | |
| 700 | 1 | |a Gòdia |D Francesc |u Grup d'Enginyeria Cellular i Bioprocés, Escola d'Enginyeria, Universitat Autònoma de Barcelona, Campus de Bellaterra, Cerdanyola del Vallès, 08193, Barcelona, Spain |4 aut | |
| 773 | 0 | |t Applied Microbiology and Biotechnology |d Springer Berlin Heidelberg |g 99/23(2015-12-01), 9935-9949 |x 0175-7598 |q 99:23<9935 |1 2015 |2 99 |o 253 | |
| 856 | 4 | 0 | |u https://doi.org/10.1007/s00253-015-6842-4 |q text/html |z Onlinezugriff via DOI |
| 898 | |a BK010053 |b XK010053 |c XK010000 | ||
| 900 | 7 | |a Metadata rights reserved |b Springer special CC-BY-NC licence |2 nationallicence | |
| 908 | |D 1 |a research-article |2 jats | ||
| 949 | |B NATIONALLICENCE |F NATIONALLICENCE |b NL-springer | ||
| 950 | |B NATIONALLICENCE |P 856 |E 40 |u https://doi.org/10.1007/s00253-015-6842-4 |q text/html |z Onlinezugriff via DOI | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Cervera |D Laura |u Grup d'Enginyeria Cellular i Bioprocés, Escola d'Enginyeria, Universitat Autònoma de Barcelona, Campus de Bellaterra, Cerdanyola del Vallès, 08193, Barcelona, Spain |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Fuenmayor |D Javier |u Grup d'Enginyeria Cellular i Bioprocés, Escola d'Enginyeria, Universitat Autònoma de Barcelona, Campus de Bellaterra, Cerdanyola del Vallès, 08193, Barcelona, Spain |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a González-Domínguez |D Irene |u Grup d'Enginyeria Cellular i Bioprocés, Escola d'Enginyeria, Universitat Autònoma de Barcelona, Campus de Bellaterra, Cerdanyola del Vallès, 08193, Barcelona, Spain |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Gutiérrez-Granados |D Sonia |u Grup d'Enginyeria Cellular i Bioprocés, Escola d'Enginyeria, Universitat Autònoma de Barcelona, Campus de Bellaterra, Cerdanyola del Vallès, 08193, Barcelona, Spain |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Segura |D Maria |u Bluebird Bio Pharmaceutical Sciences, Process Development Group, 150 2nd Street, 02141, Cambridge, MA, USA |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Gòdia |D Francesc |u Grup d'Enginyeria Cellular i Bioprocés, Escola d'Enginyeria, Universitat Autònoma de Barcelona, Campus de Bellaterra, Cerdanyola del Vallès, 08193, Barcelona, Spain |4 aut | ||
| 950 | |B NATIONALLICENCE |P 773 |E 0- |t Applied Microbiology and Biotechnology |d Springer Berlin Heidelberg |g 99/23(2015-12-01), 9935-9949 |x 0175-7598 |q 99:23<9935 |1 2015 |2 99 |o 253 | ||