Stirred bioreactors are commonly used unit operations in the pharmaceutical industry. In this study, computational fluid dynamics (CFD) was used in order to analyze the influence of the impeller configuration (Segment–Segment and Segment–Rushton impeller configurations) and the impeller rotational speed (an operational parameter) on the hydrodynamic behaviour and mixing performance of a bioreactor equipped with a double impeller. A relatively close agreement between the power values obtained from the CFD model and those measured experimentally was observed. Various parameters such as velocity profiles, stress generated by impellers due to the turbulence and velocity gradient, flow number, and mixing time were used to compare the CFD simulations. It was observed that the impeller’s RPM could change the intensity of the interaction between the impellers when a Segment–Rushton impeller was used. In general, increasing the RPM led to an increase in total power and the stress acting on the cells and to a shorter mixing time. At a constant RPM, the Segment–Rushton impeller configuration had higher total power and stress acting on cells compared to the Segment–Segment impeller configuration. At lower RPM values (i.e., 50 and 100), the Segment–Segment impeller provided a shorter mixing time. Conversely, at the highest RPM (i.e., 150) the Segment–Rushton impeller had a shorter mixing time compared to the Segment–Segment impeller; this was attributed to the high level of turbulence generated with the former impeller configuration at high RPM.

The pharmaceutical industry has witnessed exponential growth in production volumes, driven by factors such as an aging global population and the COVID-19 pandemic. To meet the demand for high product quality alongside increased productivity, there is a growing emphasis on developing innovative Fermentation Analytical Technology (FAT) and Process Analytical Technology (PAT) tools for real-time performance monitoring, modeling, measurement, and control. Building on our earlier work involving in-line monitoring of Bordetella pertussis fermentations using fluorescence spectroscopy, this review explores and compares the applications of vibrational and fluorescence spectroscopy for real-time bioprocess monitoring. We examine recent technological advancements and ongoing challenges in the field. Various spectroscopic techniques are evaluated in terms of cost-effectiveness and practical applicability, with a particular focus on in-line spectroscopy as a promising, low-cost solution for effective bioprocess monitoring.