Zeng Yu-Jing, Hsu Min-Kung, Cai Jia-Rong, Wang Hsian-Yu
International Degree Program in Animal Vaccine Technology, International College, National Pingtung University of Science and Technology, Pingtung, 91201, Taiwan.
Graduate Institute of Animal Vaccine Technology, College of Veterinary Medicine, National Pingtung University of Science and Technology, Pingtung, 91201, Taiwan.
Sci Rep. 2024 Feb 19;14(1):4092. doi: 10.1038/s41598-024-54847-7.
In the increasing demand for virus vaccines, large-scale production of safe, efficient, and economical viral antigens has become a significant challenge. High-cell-density manufacturing processes are the most commonly used to produce vaccine antigens and protein drugs. However, the cellular stress response in large-scale cell culture may directly affect host cell growth and metabolism, reducing antigen production and increasing production costs. This study provided a novel strategy of the antioxidant auxiliary system (AAS) to supply molecular hydrogen (H) into the cell culture media via proton exchange membrane (PEM) electrolysis. Integrated with a high-density cell bioreactor, the AAS aims to alleviate cellular stress response and increase viral vaccine production. In the results, the AAS stably maintained H concentration in media even in the high-air exposure tiding cell bioreactor. H treatment was shown safe to cell culture and effectively alleviated oxidative stress. In two established virus cultures models, bovine epidemic fever virus (BEFV) and porcine circovirus virus type 2 (PCV-2), were employed to verify the efficacy of AAS. The virus yield was increased by 3.7 and 2.5 folds in BEFV and PCV-2 respectively. In conclusion, the AAS-connected bioreactor effectively alleviated cellular oxidative stress and enhanced virus production in high-density cell culture.
在对病毒疫苗需求不断增加的情况下，大规模生产安全、高效且经济的病毒抗原已成为一项重大挑战。高细胞密度生产工艺是生产疫苗抗原和蛋白质药物最常用的方法。然而，大规模细胞培养中的细胞应激反应可能直接影响宿主细胞的生长和代谢，降低抗原产量并增加生产成本。本研究提供了一种抗氧化辅助系统（AAS）的新策略，通过质子交换膜（PEM）电解将分子氢（H₂）供应到细胞培养基中。与高密度细胞生物反应器相结合，AAS旨在减轻细胞应激反应并提高病毒疫苗产量。结果表明，即使在高空气暴露的潮汐式细胞生物反应器中，AAS也能稳定维持培养基中的H₂浓度。H₂处理对细胞培养安全，并有效减轻氧化应激。在两种已建立的病毒培养模型中，分别采用牛流行热病毒（BEFV）和猪圆环病毒2型（PCV - 2）来验证AAS的功效。BEFV和PCV - 2的病毒产量分别提高了3.7倍和2.5倍。总之，连接AAS的生物反应器有效减轻了细胞氧化应激，并提高了高密度细胞培养中的病毒产量。
