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José Escandell Filipa Moura Sofia B. Carvalho Ricardo J. S. Silva Ricardo Correia Antonio Roldão Patrícia Gomes-Alves Paula M. Alves 《Biotechnology and bioengineering》2023,120(9):2578-2587
The majority of recombinant adeno-associated viruses (rAAV) approved for clinical use or in clinical trials areproduced by transient transfection using the HEK293 cell line. However, this platform has several manufacturing bottlenecks at commercial scales namely, low product quality (full to empty capsid ratio <20% in most rAAV serotypes), lower productivities obtained after scale-up and the high cost of raw materials, in particular of Good Manufacturing Practice grade plasmid DNA required for transfection. The HeLa-based stable cell line rAAV production system provides a robust and scalable alternative to transient transfection systems. Nevertheless, the time required to generate the producer cell lines combined with the complexity of rAAV production and purification processes still pose several barriers to the use of this platform as a suitable alternative to the HEK293 transient transfection. In this work we streamlined the cell line development and bioprocessing for the HeLaS3-based production of rAAV. By exploring this optimized approach, producer cell lines were generated in 3-4 months, and presented rAAV2 volumetric production (bulk) > 3 × 1011 vg/mL and full to empty capsids ratio (>70%) at 2 L bioreactor scale. Moreover, the established downstream process, based on ion exchange and affinity-based chromatography, efficiently eliminated process related impurities, including the Adenovirus 5 helper virus required for production with a log reduction value of 9. Overall, we developed a time-efficient and robust rAAV bioprocess using a stable producer cell line achieving purified rAAV2 yields > 1 × 1011 vg/mL. This optimized platform may address manufacturing challenges for rAAV based medicines. 相似文献
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Khalilpour Akbar Kilic Tugba Khalilpour Saba Álvarez Mario Moisés Yazdi Iman K. 《Applied microbiology and biotechnology》2017,101(2):475-491
Applied Microbiology and Biotechnology - In the post-genome age, proteomics is receiving significant attention because they provide an invaluable source of biological structures and functions at... 相似文献
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《Nature biotechnology》2006,24(11):1300
Second-generation biologics are now entering the marketplace. 相似文献
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Oliver Spadiut Simon Rittmann Christian Dietzsch Christoph Herwig 《Engineering in Life Science》2013,13(1):88-101
In this review, we summarise recent studies that purposefully employed dynamic conditions, such as shifts, pulses, ramps and oscillations, for fast physiological strain characterisation and bioprocess development. We show the broad applicability of dynamic conditions and the various objectives that can thereby be investigated in a short time. Dynamic processes reveal information about the analysed system faster than traditional strategies, like continuous cultivations, as process parameters can directly be linked to platform and product parameters. Furthermore, we demonstrate that dynamic operations can result in increased productivity and high product quality, making this strategy a valuable tool for bioprocess development. With this review, we would like to encourage bioprocess engineers to an increased use of dynamic conditions in bioprocess development. 相似文献
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Koutinas AA Sypsas V Kandylis P Michelis A Bekatorou A Kourkoutas Y Kordulis C Lycourghiotis A Banat IM Nigam P Marchant R Giannouli M Yianoulis P 《PloS one》2012,7(4):e34350
Delignified cellulosic material has shown a significant promotional effect on the alcoholic fermentation as yeast immobilization support. However, its potential for further biotechnological development is unexploited. This study reports the characterization of this tubular/porous cellulosic material, which was done by SEM, porosimetry and X-ray powder diffractometry. The results showed that the structure of nano-tubular cellulose (NC) justifies its suitability for use in "cold pasteurization" processes and its promoting activity in bioprocessing (fermentation). The last was explained by a glucose pump theory. Also, it was demonstrated that crystallization of viscous invert sugar solutions during freeze drying could not be otherwise achieved unless NC was present. This effect as well as the feasibility of extremely low temperature fermentation are due to reduction of the activation energy, and have facilitated the development of technologies such as wine fermentations at home scale (in a domestic refrigerator). Moreover, NC may lead to new perspectives in research such as the development of new composites, templates for cylindrical nano-particles, etc. 相似文献
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Feldheim Kevin A. Jabado Rima W. Chapman Demian D. Cardeñosa Diego Maddox J. Dylan 《Molecular biology reports》2020,47(4):2669-2675
Molecular Biology Reports - Microsatellites are useful in studies of population genetics, sibship, and parentage. Here, we screened for microsatellites from multiple elasmobranch genomic libraries... 相似文献
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Thiel KA 《Nature biotechnology》2004,22(5):513-519
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Balkwill F 《Nature reviews. Immunology》2005,5(6):509-512
Immunologists need to establish a vibrant dialogue with young people. This is not only important for the continuation and progress of biomedical research, but it can also contribute to the fight against diseases such as HIV/AIDS and can help young people to make informed decisions about lifestyle, medical treatment and ethical issues. Good communication skills are crucial to any scientific career, and the lessons learned from talking with non-scientists can also be useful when writing scientific papers and grants. This article is a personal account of one scientist's experience of communicating biomedical science to young people. 相似文献
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Application of systems biology for bioprocess development 总被引:4,自引:0,他引:4
Random mutagenesis or genetic modification of an organism without consideration of its consequences to the entire system might cause unwanted changes in cellular metabolism. Systems metabolic engineering thus aims to develop strains by performing metabolic engineering within a systems biology framework, in which entire cellular networks are optimized and fermentation and downstream processes are considered at early stages. Thus, regulatory, metabolic and other cellular networks are engineered in an integrated manner. Here, we review the applications of systems biology for the development of strains and bioprocesses by means of several successful examples and, furthermore, discuss future prospects. 相似文献