Abstract
This study described a successful application of the Quality by Design (QbD) approach to pseudorabies virus (PRV) production process development in a fixed-bed bioreactor using the serum-free medium (SFM). The innovated tube-fixed-bed bioreactor was used as a scale-down model of the fixed-bed bioreactor for process development. Risk analysis was performed using Ishikawa diagram combined with failure mode effects analysis (FMEA). The comparative experiment was performed to screen proper medium for adherent African green monkey kidney (Vero) cells from three commercially available SFMs (VP-SFM, ProVERO-1 and Vero-A). The Vero-A medium showed as an outstanding one for further study. The PRV titer in harvest medium was consider as Critical Quality Attribute (CQA) and the Critical Process Parameters (CPPs) [time of infection (TOI), multiplicity of infection (MOI) and initial inoculation cell density] ranked high with risk priority number (RPN) were taken into design of experiment (DoE) methodology. Then prediction model of PRV production process was established and a robust PRV production process was explored. Under the robust setpoint conditions, the Xcell 1 L laboratory-scale fixed-bed bioreactor yielded PRV titer up to 7.87 log10 TCID50/mL at 3 dpi, which was comparable with that in the tube-fixed-bed bioreactor. Combination of the tube-fixed-bed bioreactor and QbD approach could further accelerate the development of a robust virus production process.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Abu-Absi SF, Yang L, Thompson P, Jiang C, Kandula S, Schilling B, Shukla AA (2010) Defining process design space for monoclonal antibody cell culture. Biotechnol Bioeng 106:894–905. https://doi.org/10.1002/bit.22764
Barrett PN, Mundt W, Kistner O, Howard MK (2009) Vero cell platform in vaccine production: moving towards cell culture-based viral vaccines. Expert Rev Vaccines 8:607–618. https://doi.org/10.1586/erv.09.19
Bhatia H, Read E, Agarabi C, Brorson K, Lute S, Yoon S (2016) A design space exploration for control of Critical Quality Attributes of mAb. Int J Pharm 512:242–252. https://doi.org/10.1016/j.ijpharm.2016.08.046
Chen A, Poh SL, Dietzsch C, Roethl E, Yan ML, Ng SK (2011) Serum-free microcarrier based production of replication deficient influenza vaccine candidate virus lacking NS1 using Vero cells. BMC Biotechnol 11:81. https://doi.org/10.1186/1472-6750-11-81
Frazatti-Gallina NM et al (2004) Vero-cell rabies vaccine produced using serum-free medium. Vaccine 23:511–517. https://doi.org/10.1016/j.vaccine.2004.06.014
Freuling CM, Muller TF, Mettenleiter TC (2017) Vaccines against pseudorabies virus (PrV). Vet Microbiol 206:3–9. https://doi.org/10.1016/j.vetmic.2016.11.019
Gallo-Ramirez LE, Nikolay A, Genzel Y, Reichl U (2015) Bioreactor concepts for cell culture-based viral vaccine production. Expert Rev Vaccines 14:1181–1195. https://doi.org/10.1586/14760584.2015.1067144
Huang D et al (2015) Serum-free suspension culture of MDCK cells for production of influenza H1N1 vaccines. PLoS ONE. https://doi.org/10.1371/journal.pone.0141686
Islam RS, Tisi D, Levy MS, Lye GJ (2007) Framework for the rapid optimization of soluble protein expression in Escherichia coli combining microscale experiments and statistical experimental design. Biotechnol Prog 23:785–793. https://doi.org/10.1021/bp070059a
Jordan M, Voisard D, Berthoud A, Tercier L, Kleuser B, Baer G, Broly H (2013) Cell culture medium improvement by rigorous shuffling of components using media blending. Cytotechnology 65:31–40. https://doi.org/10.1007/s10616-012-9462-1
Li X et al (2019) Quality by design-driven process development of cell culture in bioreactor for the production of foot-and-mouth veterinary vaccine. J Pharm Sci 108:2288–2295. https://doi.org/10.1016/j.xphs.2019.02.004
Liu CC, Lian WC, Butler M, Wu SC (2007) High immunogenic enterovirus 71 strain and its production using serum-free microcarrier Vero cell culture. Vaccine 25:19–24. https://doi.org/10.1016/j.vaccine.2006.06.083
Liu CC et al (2011) Purification and characterization of enterovirus 71 viral particles produced from vero cells grown in a serum-free microcarrier bioreactor system. PLoS ONE 6:e20005. https://doi.org/10.1371/journal.pone.0020005
Liu CC et al (2018) Enhancing enterovirus A71 vaccine production yield by microcarrier profusion bioreactor culture. Vaccine 36:3134–3139. https://doi.org/10.1016/j.vaccine.2017.02.042
McFerran JB, Dow C (1975) Studies on immunisation of pigs with the Bartha strain of Aujeszky’s disease virus. Res Vet Sci 19:17–22. https://doi.org/10.1016/s0034-5288(18)33548-3
Meuwly F, Ruffieux PA, Kadouri A, von Stockar U (2007) Packed-bed bioreactors for mammalian cell culture: bioprocess and biomedical applications. Biotechnol Adv 25:45–56. https://doi.org/10.1016/j.biotechadv.2006.08.004
Nagashima H, Watari A, Shinoda Y, Okamoto H, Takuma S (2013) Application of a quality by design approach to the cell culture process of monoclonal antibody production, resulting in the establishment of a design space. J Pharm Sci 102:4274–4283. https://doi.org/10.1002/jps.23744
Nie L, Hu M, Yan X, Guo T, Wang H, Zhang S, Qu H (2018) Optimization of a coupling process for insulin degludec according to a Quality by Design (QbD) paradigm. AAPS PharmSciTech 19:1–10. https://doi.org/10.1208/s12249-018-1032-1
Nie J et al (2019) Production process development of pseudorabies virus vaccine by using novel scale-down model of fixed-bed bioreactor. J Pharm Sci. https://doi.org/10.1016/j.xphs.2019.10.002
Ozturk SS, Palsson BO (1991) Examination of serum and bovine serum-albumin as shear protective agents in agitated cultures of hybridoma cells. J Biotechnol 18:13–28. https://doi.org/10.1016/0168-1656(91)90232-K
Pramod K, Tahir MA, Charoo NA, Ansari SH, Ali J (2016) Pharmaceutical product development: a quality by design approach. Int J Pharm Investig 6:129–138. https://doi.org/10.4103/2230-973X.187350
Reed LJ, Muench H (1937) A simple method of estimating fifty per cent endpoints. Am J Epidemiol 27:493–497
Rouiller Y, Perilleux A, Collet N, Jordan M, Stettler M, Broly H (2013) A high-throughput media design approach for high performance mammalian fed-batch cultures. MAbs 5:501–511. https://doi.org/10.4161/mabs.23942
Rourou S, van der Ark A, van der Velden T, Kallel H (2007) A microcarrier cell culture process for propagating rabies virus in Vero cells grown in a stirred bioreactor under fully animal component free conditions. Vaccine 25:3879–3889. https://doi.org/10.1016/j.vaccine.2007.01.086
Rourou S, van der Ark A, Majoul S, Trabelsi K, van der Velden T, Kallel H (2009) A novel animal-component-free medium for rabies virus production in Vero cells grown on Cytodex 1 microcarriers in a stirred bioreactor. Appl Microbiol Biotechnol 85:53–63. https://doi.org/10.1007/s00253-009-2064-y
Silva AC, Delgado I, Sousa MF, Carrondo MJ, Alves PM (2008) Scalable culture systems using different cell lines for the production of Peste des Petits ruminants vaccine. Vaccine 26:3305–3311. https://doi.org/10.1016/j.vaccine.2008.03.077
Slivac I, Srček VG, RadoŠević K, Kmetič I, Kniewald Z (2006) Aujeszky’s disease virus production in disposable bioreactor. J Biosci 31:363–368. https://doi.org/10.1007/bf02704109
Souza MC, Freire MS, Schulze EA, Gaspar LP, Castilho LR (2009) Production of yellow fever virus in microcarrier-based Vero cell cultures. Vaccine 27:6420–6423. https://doi.org/10.1016/j.vaccine.2009.06.023
Srcek VG, Cajavec S, Sladic D, Kniewald Z (2004) BHK 21 C13 cells for Aujeszky’s disease virus production using the multiple harvest process. Cytotechnology 45:101–106. https://doi.org/10.1007/s10616-004-2551-z
Toriniwa H, Komiya T (2007) Japanese encephalitis virus production in Vero cells with serum-free medium using a novel oscillating bioreactor. Biologicals 35:221–226. https://doi.org/10.1016/j.biologicals.2007.02.002
Wood HA, Johnston LB, Burand JP (1982) Inhibition of Autographa californica nuclear polyhedrosis virus replication in high-density Trichoplusia ni cell cultures. Virology 119:245–254. https://doi.org/10.1016/0042-6822(82)90085-x
Yuk IH et al (2006) A serum-free Vero production platform for a chimeric virus vaccine candidate. Cytotechnology 51:183–192. https://doi.org/10.1007/s10616-006-9030-7
Zhang Y, Stobbe P, Silvander CO, Chotteau V (2015) Very high cell density perfusion of CHO cells anchored in a non-woven matrix-based bioreactor. J Biotechnol 213:28–41. https://doi.org/10.1016/j.jbiotec.2015.07.006
Acknowledgements
This work was supported by the National Key Research and Development Program of China (No. 2018YFA0900804); the National Natural Science Foundation of China (No. 21878124; 31570034); the Collaborative Innovation Center of Jiangsu Modern Industrial Fermentation, the 111 Project (No. 111-2-06); TaiShan industrial Experts Programme (tscy20160307).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interests
The authors declare that they have no conflict of interest.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Nie, J., Sun, Y., Han, F. et al. Rapid process development of serum-free pseudorabies virus production with the Quality by Design approach. Cytotechnology 72, 283–293 (2020). https://doi.org/10.1007/s10616-020-00377-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10616-020-00377-9