Monitoring of Nutrients, Metabolites, IgG Titer, and Cell Densities in 10 L Bioreactors Using Raman Spectroscopy and PLS Regression Models

<b>Background:</b> Chinese hamster ovary (CHO) cell metabolism is complex, influenced by nutrients like glucose and glutamine and metabolites such as lactate. Real-time monitoring is necessary for optimizing culture conditions and ensuring consistent product quality. Raman spectroscopy has emerged as a robust process analytical technology (PAT) tool due to its non-invasive, in situ capabilities. This study evaluates Raman spectroscopy for monitoring key metabolic parameters and IgG titer in CHO cell cultures. <b>Methods:</b> Raman spectroscopy was applied to five 10 L-scale CHO cell cultures. Partial least squares (PLS) regression models were developed from four batches, including one with induced cell death, to enhance robustness. The models were validated against blind test sets. <b>Results:</b> PLS models exhibited high predictive accuracy (R² > 0.9). Glucose and IgG titer predictions were reliable (RMSEP = 0.51 g/L and 0.12 g/L, respectively), while glutamine and lactate had higher RMSEP due to lower concentrations. Specific Raman bands contributed to the specificity of glucose, lactate, and IgG models. Predictions for viable (VCD) and total cell density (TCD) were less accurate due to the absence of direct Raman signals. <b>Conclusions:</b> This study confirms Raman spectroscopy's potential for real-time, in situ bioprocess monitoring without manual sampling. Chemometric analysis enhances model robustness, supporting automated control systems. Raman data could enable continuous feedback regulation of critical nutrients like glucose, ensuring consistent critical quality attributes (CQAs) in biopharmaceutical production.