Föller Stefanie, Regett Niklas, Lataster Levin, Radziwill Gerald, Takors Ralf
Institute of Biochemical Engineering, University of Stuttgart, Allmandring 31, 70569, Stuttgart, Germany.
Institute of Biology II, University of Freiburg, 79098, Freiburg, Germany.
Appl Microbiol Biotechnol. 2024 Dec 5;108(1):530. doi: 10.1007/s00253-024-13363-4.
Research for biopharmaceutical production processes with mammalian cells steadily aims to enhance the cell-specific productivity as a means for optimizing total productivities of bioreactors. Whereas current technologies such as pH, temperature, and osmolality shift require modifications of the cultivation medium, the use of optogenetic switches in recombinant producer cells might be a promising contact-free alternative. However, the proper application of optogenetically engineered cells requires a detailed understanding of basic cellular responses of cells that do not yet contain the optogenetic switches. The knowhow of ideal light exposure to enable the optimum use of related approaches is missing so far. Consequently, the current study set out to find optimum conditions for IgG1 producing Chinese hamster ovary (CHO) cells which were exposed to blue LED light. Growth characteristics, cell-specific productivity using enzyme-linked immunosorbent assay, as well as cell cycle distribution using flow cytometry were analyzed. Whereas too harsh light exposure causes detrimental growth effects that could be compensated with antioxidants, a surprising boost of cell-specific productivity by 57% occurred at optimum high light doses. The increase coincided with an increased number of cells in the G1 phase of the cell cycle after 72 h of illumination. The results present a promising new approach to boost biopharmaceutical productivity of mammalian cells simply by proper light exposure without any further optogenetic engineering. KEY POINTS: • Blue LED light hinders growth in CHO DP-12 cells • Antioxidants protect to a certain degree from blue light effects • Illumination with blue LED light raises cell-specific productivity.
利用哺乳动物细胞进行生物制药生产工艺的研究一直致力于提高细胞特异性生产力，以此作为优化生物反应器总生产力的一种手段。虽然当前诸如pH值、温度和渗透压变化等技术需要对培养基进行调整，但在重组生产细胞中使用光遗传学开关可能是一种有前景的非接触式替代方法。然而，光遗传学工程细胞的正确应用需要详细了解尚未包含光遗传学开关的细胞的基本细胞反应。到目前为止，尚缺乏关于理想光照以实现相关方法最佳利用的技术诀窍。因此，当前研究着手寻找暴露于蓝色发光二极管（LED）光下的产IgG1的中国仓鼠卵巢（CHO）细胞的最佳条件。分析了生长特性、使用酶联免疫吸附测定法测定的细胞特异性生产力以及使用流式细胞术测定的细胞周期分布。虽然过于强烈的光照会导致有害的生长效应，而抗氧化剂可以对此进行补偿，但在最佳高光剂量下，细胞特异性生产力会惊人地提高57%。这种增加与光照72小时后细胞周期G1期细胞数量的增加相吻合。结果表明，仅通过适当的光照而无需任何进一步的光遗传学工程，就能提高哺乳动物细胞生物制药生产力，这是一种有前景的新方法。要点：• 蓝色LED光阻碍CHO DP - 12细胞生长 • 抗氧化剂在一定程度上保护细胞免受蓝光影响 • 用蓝色LED光照提高细胞特异性生产力
