tetano
Editor, Senior Moderator
J Biol Eng. 2019 Oct 23;13:78. doi: 10.1186/s13036-019-0206-z. eCollection 2019. [h=1]Process development for pandemic influenza VLP vaccine production using a baculovirus expression system.[/h]
Lai CC[SUP]1,[/SUP][SUP]2[/SUP], Cheng YC[SUP]1[/SUP], Chen PW[SUP]1[/SUP], Lin TH[SUP]1,[/SUP][SUP]2[/SUP], Tzeng TT[SUP]1[/SUP], Lu CC[SUP]1[/SUP], Lee MS[SUP]1[/SUP], Hu AY[SUP]1[/SUP].
[h=3]Author information[/h] 1 National Institute of Infectious Diseases and Vaccinology, NHRI, 35 Keyan Road, Zhunan, Miaoli County, 35053 Taiwan. 2 2College of Life Science, National Tsing Hua University, 101, Section 2, Kuang-Fu Road, Hsinchu, 30013 Taiwan.
[h=3]Abstract[/h] [h=4]Background:[/h] Influenza viruses cause hundreds of thousands of respiratory diseases worldwide each year, and vaccination is considered the most effective approach for preventing influenza annual epidemics or pandemics. Since 1950, chicken embryonated eggs have been used as the main method for producing seasonal influenza vaccines. However, this platform has the main drawback of a lack of scale-up flexibility, and thus, egg-based vaccine manufacturers cannot supply sufficient doses within a short period for use for pandemic prevention. As a result, strategies for reducing the manufacturing time and increasing production capacity are urgently needed. Non-virion vaccine methods have been considered an alternative strategy against an influenza pandemic, and the purpose of maintaining an immunogenic capsule structure with infectious properties appears to be met by the virus-like particle (VLP) platform.
[h=4]Results:[/h] An influenza H7N9-TW VLP production platform using insect cells, which included the expression of hemagglutinin (HA), NA, and M1 proteins, was established. To scale up H7N9-TW VLP production, several culture conditions were optimized to obtain a higher production yield. A high level of dissolved oxygen (DO) could be critical to H7N9-TW VLP production. If the DO was maintained at a high level, the HA titer obtained in the spinner flask system with ventilation was similar to that obtained in a shake flask. In this study, the HA titer in a 5-L bioreactor with a well-controlled DO level was substantially improved by 128-fold (from 4 HA units (HAU)/50 μL to 512 HAU/50 μL).
[h=4]Conclusions:[/h] In this study, a multigene expression platform and an effective upstream process were developed. Notably, a high H7N9-TW VLP yield was achieved using a two-step production strategy while a high DO level was maintained. The upstream process, which resulted in high VLP titers, could be further used for large-scale influenza VLP vaccine production.
? The Author(s). 2019.
[h=4]KEYWORDS:[/h] Dissolved oxygen; Influenza vaccines; Virus-like particle
PMID: 31666806 PMCID: PMC6813129 DOI: 10.1186/s13036-019-0206-z
Lai CC[SUP]1,[/SUP][SUP]2[/SUP], Cheng YC[SUP]1[/SUP], Chen PW[SUP]1[/SUP], Lin TH[SUP]1,[/SUP][SUP]2[/SUP], Tzeng TT[SUP]1[/SUP], Lu CC[SUP]1[/SUP], Lee MS[SUP]1[/SUP], Hu AY[SUP]1[/SUP].
[h=3]Author information[/h] 1 National Institute of Infectious Diseases and Vaccinology, NHRI, 35 Keyan Road, Zhunan, Miaoli County, 35053 Taiwan. 2 2College of Life Science, National Tsing Hua University, 101, Section 2, Kuang-Fu Road, Hsinchu, 30013 Taiwan.
[h=3]Abstract[/h] [h=4]Background:[/h] Influenza viruses cause hundreds of thousands of respiratory diseases worldwide each year, and vaccination is considered the most effective approach for preventing influenza annual epidemics or pandemics. Since 1950, chicken embryonated eggs have been used as the main method for producing seasonal influenza vaccines. However, this platform has the main drawback of a lack of scale-up flexibility, and thus, egg-based vaccine manufacturers cannot supply sufficient doses within a short period for use for pandemic prevention. As a result, strategies for reducing the manufacturing time and increasing production capacity are urgently needed. Non-virion vaccine methods have been considered an alternative strategy against an influenza pandemic, and the purpose of maintaining an immunogenic capsule structure with infectious properties appears to be met by the virus-like particle (VLP) platform.
[h=4]Results:[/h] An influenza H7N9-TW VLP production platform using insect cells, which included the expression of hemagglutinin (HA), NA, and M1 proteins, was established. To scale up H7N9-TW VLP production, several culture conditions were optimized to obtain a higher production yield. A high level of dissolved oxygen (DO) could be critical to H7N9-TW VLP production. If the DO was maintained at a high level, the HA titer obtained in the spinner flask system with ventilation was similar to that obtained in a shake flask. In this study, the HA titer in a 5-L bioreactor with a well-controlled DO level was substantially improved by 128-fold (from 4 HA units (HAU)/50 μL to 512 HAU/50 μL).
[h=4]Conclusions:[/h] In this study, a multigene expression platform and an effective upstream process were developed. Notably, a high H7N9-TW VLP yield was achieved using a two-step production strategy while a high DO level was maintained. The upstream process, which resulted in high VLP titers, could be further used for large-scale influenza VLP vaccine production.
? The Author(s). 2019.
[h=4]KEYWORDS:[/h] Dissolved oxygen; Influenza vaccines; Virus-like particle
PMID: 31666806 PMCID: PMC6813129 DOI: 10.1186/s13036-019-0206-z