Process analytical technology tools for perfusion cell culture

Abstract

During cell cultivation processes for the production of biopharmaceuticals, good process performance and good product quality can be ensured by online monitoring of critical process parameters (e.g. temperature, pH, or dissolved oxygen). These data can be used in real‐time for process control, as suggested by the process analytical technology (PAT) initiative. Today, solutions for real‐time monitoring of parameters such as concentrations of cells, main nutrients, and metabolism by‐products are developing, but applications of these more complex tools in industrial settings are still limited. Here, we evaluated the use of dielectric spectroscopy (DS) and near‐infrared spectroscopy (NIRS) as PAT tools for a perfusion PER.C6® cultivation process. We showed that DS enabled predictions of viable cell density from the cultivation vessel, with a root mean square error of prediction (RMSEP) of 4.4% of the calibration range. Additionally, predictions of glucose and lactate concentrations from the cultivation vessel (RMSEP of 10 and 14%, respectively) and from the perfusion stream (RMSEP of 12 and 10%, respectively) were achieved with NIRS. We also showed that the perfusion stream offers great opportunities for noninvasive, yet frequent process monitoring. Accurate online monitoring of critical process parameters with PAT tools is the essential first step toward increased control of process output.

Enabling PAT in insect cell bioprocesses: In situ monitoring of recombinant adeno‐associated virus production by fluorescence spectroscopy

“…Several spectroscopic techniques have been used for cell culture process and virus production monitoring, with Raman (Rangan et al, 2018;Santos, Kessler, Salou, Menezes, & Peinado, 2018;Webster, Hadley, Hilliard, Jaques, & Mason, 2018), near-infrared (Mercier et al, 2015;Rowland-Jones, van den Berg, Racher, Martin, & Jaques, 2017), dielectric (Kroll, Stelzer, & Herwig, 2017;Mercier et al, 2015;Nikolay, Léon, Schwamborn, Genzel, & Reichl, 2018;Petiot, Ansorge, Rosa-Calatrava, & Kamen, 2016) and fluorescence spectroscopy (Karakach et al, 2018;Schwab & Hesse, 2017) being the most widely used. All possess characteristics desirable for PAT: spectroscopic techniques are noninvasive, nondestructive, and able to provide rapid information from several components simultaneously (Ohadi, Aghamohseni, Legge, & Budman, 2014;Ohadi, Legge, & Budman, 2015;Rowland-Jones et al, 2017;Teixeira et al, 2011Teixeira et al, , 2009, including product quality attributes (Chopda, Pathak, Batra, Gomes, & Rathore, 2017;Li et al, 2019).…”

Decision Tree‐PLS (DT‐PLS) algorithm for the development of process: Specific local prediction models

“…For the broad and automated integration of this technology into the R&D and manufacturing ecosystem, data‐ and knowledge‐driven model‐based solutions play a key role. Data‐driven methods are centered on the generated process data sets and target enhanced process understanding and prediction through visualization, correlation analysis, and regression techniques . Given the complexity and variety of bioprocess data, a customization of data‐driven tools on the basis of process specificities and analysis expertise is crucial for their consistent utilization.…”

“…Datadriven methods are centered on the generated process data sets and target enhanced process understanding and prediction through visualization, correlation analysis, and regression techniques. [4][5][6][7][8][9][10] Given the complexity and variety of bioprocess data, a customization of data-driven tools on the basis of process specificities and analysis expertise is crucial for their consistent utilization.…”

A novel scale‐down mimic of perfusion cell culture using sedimentation in an automated microbioreactor (SAM)

“…The alternating flow filtration system (ATF) (Repligen, MA) and CARR Centritech separation system (Pneumatic scale angelus, OH) have become established as the most commonly used methods . These new cell retention devices in combination with other technologies like single‐use processing or advanced process control by PAT technology also decreased the technical complexity and lowered the associated failure rate . The lack of appropriate scale‐down systems is addressed within this work.…”

“…[7][8][9] These new cell retention devices in combination with other technologies like single-use processing or advanced process control by PAT technology also decreased the technical complexity and lowered the associated failure rate. 10 The lack of appropriate scale-down systems is addressed within this work.…”