Chromatography is an integral step in downstream bioprocessing, and buffered mobile-phase properties such as ionic strength and pH are often critical process parameters for protein purification. The interactions between the mobile and stationary phases can affect chromatographic performance, and thus the separation profile between product and impurities. Product recovery, purity, and throughput are often marred by the variability in buffer preparation during production-scale bioprocessing. The popular approach to biological production typically relies on postchromatographic adjustments (e.g., fractionation controlled by ultraviolet (UV) signal and segregation of out-of-specification product through quality-control testing) to achieve the desired product quality. Although the US Food and Drug Administration accepts this traditional practice, this post-process approach to quality control is time-consuming, increases production cost, and does not provide the process understanding and control from the quality-by-design (QbD) approach that FDA promotes.
Process analytical technology
In accordance with FDA's Pharmaceutical CGMPs for the 21st Century initiatives, companies should employ process monitoring and control strategies to streamline biopharmaceutical manufacturing. Real-time process knowledge can help manufacturers avoid excessive rework and discarded product. To achieve this standard of quality, FDA highly recommends the implementation of process analytical technology (PAT) to facilitate a QbD approach. According to FDA, the working definition of PAT is "a system for designing, analyzing, and controlling manufacturing through timely measurements (i.e., during process) of critical quality and performance attributes of raw and in-process materials and processes with the goal of ensuring final product quality" (1).Many modern technniques for process analysis such as pH measurement, near-infrared spectroscopy, Raman spectroscopy, refractive-index changes, humidity movement, UV monitoring, conductivity detection, and dissolved-oxygen analysis are commercially available for different applications. These analytical tools can measure physical and chemical properties of raw materials directly in real time, in contrast with techniques such as flow-meter monitoring, which only makes indirect measurements.
Applications of PAT in downstream bioprocessing