在单一的多分室生物反应器中模拟CHO大规模效应:一种新的方法来访问扩大规模的行为
Lena Gaugler1, Sebastian Hofmann2, Michael Schlüter2
1Institute of Biochemical Engineering, University of Stuttgart, Stuttgart, Germany.
Biotechnology and bioengineering
|January 9, 2024
概括
这项研究引入了一种新的生物反应器系统,以模仿大规模生物制药生产的挑战. 它揭示了输入功率,而不是异质性,主要影响缩放期间的糖化.
科学领域:
- 生物制药制造业 生物制药制造业
- 细胞培养技术 细胞培养技术
- 过程扩大规模的升级.
背景情况:
- 生物制药生产的扩大规模可能会导致由于梯度和异质性而导致不可预测的性能损失.
- 现有的方法很难预测或解释实验室和商业尺度之间的不一致性.
研究的目的:
- 引入和验证一种新的缩放式反应器系统,即单个多分室生物反应器 (SMCB),用于研究哺乳动物细胞培养异质性.
- 在扩展过程中区分输入功率和异质性对细胞培养性能和产品质量的影响.
主要方法:
- 在SMCB中进行了中国仓鼠卵巢 (CHO) 食批量培养,模拟了长混合时间和溶氧 (DO) 异质性等大规模条件.
- 与参考生物反应器 (REFB) 设置相比较的SMCB条件 (20.4-1.5W m-3).
- 分析了细胞生长,基质吸收,乳酸积累和甘氨酸模式.
主要成果:
- SMCB条件导致乳酸积累增加 (高达87%),葡萄糖吸收增加,活细胞度降低,这是大规模表现的特征.
- 产品质量属性,特别是银河化程度,主要受到输入功率变化的影响,而不是诱导的异质性.
- SMCB成功地模仿了大规模的异质性和性能问题.
结论:
- SMCB是了解生物制药过程扩大规模的挑战和预测细胞系特异性问题在早期发展的宝贵工具.
- 输入功率是影响缩放过程中的糖化的一个关键因素,可能比异质性更重要.
- 该系统为减轻大规模生物生产中的性能损失提供了新的见解.
相关概念视频
01:29Bioreactor Design and Operational System
Bioreactors are engineered vessels designed to cultivate microorganisms under controlled conditions for industrial bioprocessing. They maintain sterility and allow precise regulation of pH, temperature, oxygen, and nutrient levels to optimize microbial growth and metabolite production. Bioreactors range from small laboratory units of 1 liter to industrial systems holding up to 500,000 liters, though only about 75% of their volume is actively used for fermentation. The remaining headspace...
01:18Bioreactor Controls-II
In aerobic fermentations, oxygen is vital for microbial growth and metabolite production. Since air comprises only about 20% oxygen and the gas is poorly soluble in water—just 9 ppm at 20°C—supplying sufficient oxygen becomes a critical challenge, especially in high-demand processes like yeast growth or citric acid production. Even a fully saturated broth may offer only a few seconds of oxygen availability.To address this, sterile or scrubbed air is introduced into the fermentor via a sparger...
01:22Bioreactor Controls-III
Strain improvement is a foundational strategy in industrial microbiology aimed at maximizing microbial productivity, particularly because natural isolates typically yield commercially valuable products in very low concentrations. Although optimizing the culture medium and environmental conditions can improve yields, these adjustments are inherently limited by the organism’s genetic potential. As a result, the focus shifts toward genetic modifications to enhance biosynthetic capacity. The...
01:28Methods of Medium Optimization
Optimizing growth media enhances microbial proliferation and maximizes product yield. Statistical experimental design methodologies provide structured and reproducible approaches, offering progressively higher levels of robustness and efficiency.The One-Factor-at-a-Time (OFAT) MethodThe One-Factor-at-a-Time (OFAT) method involves adjusting a single variable while keeping all others constant. However, it cannot detect interactions between variables, often leading to suboptimal outcomes when...
01:14Scale-Up Processes
The scale-up of microbial fermentation processes is essential in industrial biotechnology, allowing the transition from laboratory-scale experiments to commercial-scale production while aiming to maintain product yield and quality. This process requires meticulous adjustment of equipment design, process parameters, and contamination control strategies to accommodate increasing culture volumes.At the laboratory scale, cultures are typically maintained in 1 to 10-liter glass or autoclavable...
01:27Upstream Processing
Upstream processing represents a critical phase in biomanufacturing, wherein biological systems such as microorganisms, mammalian cells, or insect cells are cultivated to produce therapeutic proteins, vaccines, enzymes, or other biologically derived products. This phase encompasses all steps from the selection and genetic manipulation of the production organism to the cultivation of cells in bioreactors under tightly controlled environmental conditions.Host Selection and Genetic OptimizationThe...