caa a22 4500 605502927 CHVBK 20210128100608.0 cr unu---uuuuu 210128e20150901xx s 000 0 eng 10.1007/s00253-015-6636-8 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6636-8 Production of canine adenovirus type 2 in serum-free suspension cultures of MDCK cells [Elektronische Daten] [R. Castro, P. Fernandes, T. Laske, M. Sousa, Y. Genzel, K. Scharfenberg, P. Alves, A. Coroadinha] 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. Springer-Verlag Berlin Heidelberg, 2015 Canine adenovirus nationallicence MDCK nationallicence Serum-free media nationallicence Suspension cultures nationallicence Process monitoring nationallicence Castro R. iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2781-901, Oeiras, Portugal aut Fernandes P. iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2781-901, Oeiras, Portugal aut Laske T. iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2781-901, Oeiras, Portugal aut Sousa M. iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2781-901, Oeiras, Portugal aut Genzel Y. Bioprocess Engineering, Max Planck Institute for Dynamics of Complex Technical Systems, Sandtorstr. 1, 39106, Magdeburg, Germany aut Scharfenberg K. University of Applied Sciences Emden/Leer, Constantiaplatz 4, 26723, Emden, Germany aut Alves P. iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2781-901, Oeiras, Portugal aut Coroadinha A. iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2781-901, Oeiras, Portugal aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/17(2015-09-01), 7059-7068 0175-7598 99:17<7059 2015 99 253 https://doi.org/10.1007/s00253-015-6636-8 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6636-8 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Castro R. iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2781-901, Oeiras, Portugal aut NATIONALLICENCE 700 1- Fernandes P. iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2781-901, Oeiras, Portugal aut NATIONALLICENCE 700 1- Laske T. iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2781-901, Oeiras, Portugal aut NATIONALLICENCE 700 1- Sousa M. iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2781-901, Oeiras, Portugal aut NATIONALLICENCE 700 1- Genzel Y. Bioprocess Engineering, Max Planck Institute for Dynamics of Complex Technical Systems, Sandtorstr. 1, 39106, Magdeburg, Germany aut NATIONALLICENCE 700 1- Scharfenberg K. University of Applied Sciences Emden/Leer, Constantiaplatz 4, 26723, Emden, Germany aut NATIONALLICENCE 700 1- Alves P. iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2781-901, Oeiras, Portugal aut NATIONALLICENCE 700 1- Coroadinha A. iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2781-901, Oeiras, Portugal aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/17(2015-09-01), 7059-7068 0175-7598 99:17<7059 2015 99 253