Institute of Biochemical Engineering, University of Stuttgart, Stuttgart, Germany.
Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach, Germany.
Biotechnol Bioeng. 2019 May;116(5):951-960. doi: 10.1002/bit.26926. Epub 2019 Jan 29.
Perfusion processes are an emerging alternative to common fed-batch processes in the growing biopharmaceutical industry. However, the challenge of maintaining high cell-specific productivities remains. In this study, glucose limitation was applied to two perfusion steady states and compared with a third steady state without any detectable limitation. The metabolic phenotype was enhanced under glucose limitation with a decrease of 30% in glucose uptake and 75% in lactate formation. Cell-specific productivities were substantially improved by 50%. Remarkably, the productivities showed a strong correlation to respiratory adenosine triphosphate (ATP) supply. As less reduced nicotinamide adenine dinucleotide (NADH) remained in the cytosol, the ATP generation from oxidative phosphorylation was increased by almost 30%. Consequently, the efficiency of carbon metabolism and the resulting respiratory ATP supply was crucial for maintaining the highly productive cellular state. This study highlights that glucose limitation can be used for process intensification in perfusion cultures as ATP generation via respiration is significantly increased, leading to elevated productivities.
灌流工艺是在不断发展的生物制药行业中替代常见分批补料工艺的新兴方法。然而，保持高细胞特异性生产率仍然是一个挑战。在这项研究中，葡萄糖限制应用于两种灌流稳态，并与第三种无明显限制的稳态进行了比较。在葡萄糖限制下，代谢表型得到增强，葡萄糖摄取减少 30%，乳酸形成减少 75%。细胞特异性生产率提高了 50%。值得注意的是，生产率与呼吸三磷酸腺苷 (ATP) 供应有很强的相关性。由于胞质中还原型烟酰胺腺嘌呤二核苷酸 (NADH) 减少，氧化磷酸化产生的 ATP 增加了近 30%。因此，碳代谢的效率和由此产生的呼吸 ATP 供应对于维持高生产力的细胞状态至关重要。这项研究强调，葡萄糖限制可用于灌流培养中的工艺强化，因为通过呼吸产生的 ATP 显著增加，从而提高生产率。
