A scalable, integrated downstream process for production of a recombinant measles virus-vectored vaccine.

Autor: Steppert P; University of Natural Resources and Life Sciences, Vienna, Department of Biotechnology, Institute of Bioprocess Science and Engineering, Muthgasse 18, 1190 Vienna, Austria; Acib - Austrian Centre of Industrial Biotechnology, 1190 Vienna, Austria., Mosor M; Acib - Austrian Centre of Industrial Biotechnology, 1190 Vienna, Austria., Stanek L; Acib - Austrian Centre of Industrial Biotechnology, 1190 Vienna, Austria., Burgstaller D; Acib - Austrian Centre of Industrial Biotechnology, 1190 Vienna, Austria., Palmberger D; Acib - Austrian Centre of Industrial Biotechnology, 1190 Vienna, Austria., Preinsperger S; Acib - Austrian Centre of Industrial Biotechnology, 1190 Vienna, Austria., Pereira Aguilar P; Acib - Austrian Centre of Industrial Biotechnology, 1190 Vienna, Austria., Müllner M; Themis Bioscience GmbH, 1190 Vienna, Austria a subsidiary of Merck & Co., Inc, Kenilworth, NJ, USA., Csar P; Themis Bioscience GmbH, 1190 Vienna, Austria a subsidiary of Merck & Co., Inc, Kenilworth, NJ, USA., Jungbauer A; University of Natural Resources and Life Sciences, Vienna, Department of Biotechnology, Institute of Bioprocess Science and Engineering, Muthgasse 18, 1190 Vienna, Austria; Acib - Austrian Centre of Industrial Biotechnology, 1190 Vienna, Austria. Electronic address: alois.jungbauer@boku.ac.at.
Jazyk: angličtina
Zdroj: Vaccine [Vaccine] 2022 Feb 23; Vol. 40 (9), pp. 1323-1333. Date of Electronic Publication: 2022 Jan 31.
DOI: 10.1016/j.vaccine.2022.01.004
Abstrakt: Purification of very large and complex, enveloped viruses, such as measles virus is very challenging, it must be performed in a closed system because the final product cannot be sterile filtered and often loss of virus titer and poor product purity has been observed. We developed a purification process where the clarified and endonuclease treated culture supernatant is loaded on a restricted access chromatography medium where small impurities are bound and the virus is collected in the flow-through, which is then concentrated, and buffer exchanged by ultra/diafiltration. Up to 98.5% of host cell proteins could be captured by direct loading of clarified and endonuclease treated cell culture supernatant. Reproducible process performance and scalability of the chromatography step were demonstrated from small to pilot scale, including loading volumes from 50 mL up to 9 L. A 10-fold virus concentration was achieved by the ultrafiltration using a 100 kDa flat-sheet membrane. The order of individual process steps had a large impact on the virus infectivity and total process yields. The developed process maintained virus infectivity and is twice as fast as the traditional process train, where concentration is performed before loading on the chromatography column. Capturing impurities by the restricted access medium makes it a platform purification process with a high flexibility, which can be easily and quickly adapted to other vectors based on the measles virus vector platform.
Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Csar reports personal fees from Themis Bioscience GmbH, during the conduct of the study; In addition, Csar has a patent EP3581646 pending, and a patent PCT/EP2019/065670 pending. Dr. Burgstaller reports grants and personal fees from acib GmbH, personal fees and non-financial support from Themis Bioscience GmbH, during the conduct of the study; Dr. Palmberger reports personal fees from Themis Bioscience GmbH, outside the submitted work; In addition, Dr. Palmberger has a patent PCT/EP2019/065670 pending. Dr. Jungbauer reports personal fees from University of Natural Resources and Life Sciences, Vienna, during the conduct of the study; In addition, Dr. Jungbauer has a patent PCT/EP2019/065670 pending. Mosor reports grants and personal fees from acib GmbH, non-financial support from Themis Bioscience GmbH. Dr. Müllner reports personal fees from Themis Bioscience GmbH, outside the submitted work; In addition, Dr. Müllner has a patent PCT/EP2019/065670 pending. Preinsperger reports personal fees and non-financial support from Themis Bioscience GmbH, grants, personal fees and non-financial support from Acib GmbH, during the conduct of the study; Stanek MSc. reports grants and personal fees from acib GmbH, personal fees and non-financial support from Themis Bioscience GmbH, during the conduct of the study; Dr. Steppert reports grants and personal fees from acib GmbH, personal fees and non-financial support from Themis Bioscience GmbH, during the conduct of the study; In addition, Dr. Steppert has a patent PCT/EP2019/065670 pending.
(Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)
Databáze: MEDLINE
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