NON-INVASIVE REAL-TIME MONITORING OF GLUCOSE AND LACTATE BY NIR SPECTROSCOPY DURING PERFUSION CHO CULTURE

摘要

Near-infrared spectroscopy (NIR) has been used to monitor glycerol and methanol, non-invasively during cultures of Pichia pastoris. In this work, glucose and lactate were measured in Chinese Hamster Ovary (CHO) perfusion culture, in real time, using an online advanced NIR monitor and a reference offline biochemical analyzer. The 1.8-L culture started in the batch phase at 4 g/L glucose with 0.3 × 10~6 cells/mL and reached 1.5 × 10~6 cells/mL after 90 hours. Perfusion was then initiated and conducted for 10 days at 0.7 wd, using a spiral membrane-less microfluidic device. The maximum cell concentration was 4 ×10~6 cells/mL at 160 hours and was maintained until the termination of the experiment. Online and offline trends were similar (Figure 1). Final concentrations of glucose and lactate were 2.2 g/L and 1.8 g/L, respectively. High performance liquid chromatography (HPLC) was used to measure IgG_1.