Production of canine adenovirus type 2 in serum-free suspension cultures of MDCK cells
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| 024 | 7 | 0 | |a 10.1007/s00253-015-6636-8 |2 doi |
| 035 | |a (NATIONALLICENCE)springer-10.1007/s00253-015-6636-8 | ||
| 245 | 0 | 0 | |a Production of canine adenovirus type 2 in serum-free suspension cultures of MDCK cells |h [Elektronische Daten] |c [R. Castro, P. Fernandes, T. Laske, M. Sousa, Y. Genzel, K. Scharfenberg, P. Alves, A. Coroadinha] |
| 520 | 3 | |a The potential of adherent Madin Darby Canine Kidney (MDCK) cells for the production of influenza viruses and canine adenovirus type 2 (CAV-2) for vaccines or gene therapy approaches has been shown. Recently, a new MDCK cell line (MDCK.SUS2) that was able to grow in suspension in a fully defined system was established. In this work, we investigated whether the new MDCK.SUS2 suspension cell line is suitable for the amplification of CAV-2 under serum-free culture conditions. Cell growth performance and CAV-2 production were evaluated in three serum-free media: AEM, SMIF8, and EXCELL MDCK. CAV-2 production in shake flasks was maximal when AEM medium was used, resulting in an amplification ratio of infectious particles (IP) of 142 IP out/IP in and volumetric and cell-specific productivities of 2.1 × 108 IP/mL and 482 IP/cell, respectively. CAV-2 production was further improved when cells were cultivated in a 0.5-L stirred tank bioreactor. To monitor infection and virus production, cells were analyzed by flow cytometry. A correlation between the side scatter measurement and CAV-2 productivity was found, which represents a key feature to determine the best harvesting time during process development of gene therapy vectors that do not express reporter genes. This work demonstrates that MDCK.SUS2 is a suitable cell substrate for CAV-2 production, constituting a step forward in developing a production process transferable to industrial scales. This could allow for the production of high CAV-2 titers either for vaccination or for gene therapy purposes. | |
| 540 | |a Springer-Verlag Berlin Heidelberg, 2015 | ||
| 690 | 7 | |a Canine adenovirus |2 nationallicence | |
| 690 | 7 | |a MDCK |2 nationallicence | |
| 690 | 7 | |a Serum-free media |2 nationallicence | |
| 690 | 7 | |a Suspension cultures |2 nationallicence | |
| 690 | 7 | |a Process monitoring |2 nationallicence | |
| 700 | 1 | |a Castro |D R. |u iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2781-901, Oeiras, Portugal |4 aut | |
| 700 | 1 | |a Fernandes |D P. |u iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2781-901, Oeiras, Portugal |4 aut | |
| 700 | 1 | |a Laske |D T. |u iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2781-901, Oeiras, Portugal |4 aut | |
| 700 | 1 | |a Sousa |D M. |u iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2781-901, Oeiras, Portugal |4 aut | |
| 700 | 1 | |a Genzel |D Y. |u Bioprocess Engineering, Max Planck Institute for Dynamics of Complex Technical Systems, Sandtorstr. 1, 39106, Magdeburg, Germany |4 aut | |
| 700 | 1 | |a Scharfenberg |D K. |u University of Applied Sciences Emden/Leer, Constantiaplatz 4, 26723, Emden, Germany |4 aut | |
| 700 | 1 | |a Alves |D P. |u iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2781-901, Oeiras, Portugal |4 aut | |
| 700 | 1 | |a Coroadinha |D A. |u iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2781-901, Oeiras, Portugal |4 aut | |
| 773 | 0 | |t Applied Microbiology and Biotechnology |d Springer Berlin Heidelberg |g 99/17(2015-09-01), 7059-7068 |x 0175-7598 |q 99:17<7059 |1 2015 |2 99 |o 253 | |
| 856 | 4 | 0 | |u https://doi.org/10.1007/s00253-015-6636-8 |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/s00253-015-6636-8 |q text/html |z Onlinezugriff via DOI | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Castro |D R. |u iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2781-901, Oeiras, Portugal |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Fernandes |D P. |u iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2781-901, Oeiras, Portugal |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Laske |D T. |u iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2781-901, Oeiras, Portugal |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Sousa |D M. |u iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2781-901, Oeiras, Portugal |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Genzel |D Y. |u Bioprocess Engineering, Max Planck Institute for Dynamics of Complex Technical Systems, Sandtorstr. 1, 39106, Magdeburg, Germany |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Scharfenberg |D K. |u University of Applied Sciences Emden/Leer, Constantiaplatz 4, 26723, Emden, Germany |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Alves |D P. |u iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2781-901, Oeiras, Portugal |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Coroadinha |D A. |u iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2781-901, Oeiras, Portugal |4 aut | ||
| 950 | |B NATIONALLICENCE |P 773 |E 0- |t Applied Microbiology and Biotechnology |d Springer Berlin Heidelberg |g 99/17(2015-09-01), 7059-7068 |x 0175-7598 |q 99:17<7059 |1 2015 |2 99 |o 253 | ||