Development of generic metabolic Raman calibration models using solution titration in aqueous phase and data augmentation for in‐line cell culture analysis

Abstract This study presents a novel approach for developing generic metabolic Raman calibration models for in‐line cell culture analysis using glucose and lactate stock solution titration in an aqueous phase and data augmentation techniques. First, a successful set‐up of the titration method was achieved by adding glucose or lactate solution at several different constant rates into the aqueous phase of a bench‐top bioreactor. Subsequently, the in‐line glucose and lactate concentration were calculated and interpolated based on the rate of glucose and lactate addition, enabling data augmentation and enhancing the robustness of the metabolic calibration model. Nine different combinations of spectra pretreatment, wavenumber range selection, and number of latent variables were evaluated and optimized using aqueous titration data as training set and a historical cell culture data set as validation and prediction set. Finally, Raman spectroscopy data collected from 11 historical cell culture batches (spanning four culture modes and scales ranging from 3 to 200 L) were utilized to predict the corresponding glucose and lactate values. The results demonstrated a high prediction accuracy, with an average root mean square errors of prediction of 0.65 g/L for glucose, and 0.48 g/L for lactate. This innovative method establishes a generic metabolic calibration model, and its applicability can be extended to other metabolites, reducing the cost of deploying real‐time cell culture monitoring using Raman spectroscopy in bioprocesses.