Control of IgG glycosylation by in situ and real‐time estimation of specific growth rate of CHO cells cultured in bioreactor

Abstract

The cell‐specific growth rate (µ) is a critical process parameter for antibody production processes performed by animal cell cultures, as it describes the cell growth and reflects the cell physiological state. When there are changes in these parameters, which are indicated by variations of µ, the synthesis and the quality of antibodies are often affected. Therefore, it is essential to monitor and control the variations of µto assure the antibody production and achieve high product quality. In this study, a novel approach for on‐line estimation of µ was developed based on the process analytical technology initiative by using an in situ dielectric spectroscopy. Critical moments, such as significant µ decreases, were successfully detected by this method, in association with changes in cell physiology as well as with an accumulation of nonglycosylated antibodies. Thus, this method was used to perform medium renewals at the appropriate time points, maintaining the values of µ close to its maximum. Using this method, we demonstrated that the physiological state of cells remained stable, the quantity and the glycosylation quality of antibodies were assured at the same time, leading to better process performances compared with the reference feed‐harvest cell cultures carried out by using off‐line nutrient measurements.

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). Both Raman and fluorescence have proven worthy for monitoring recombinant protein production and cell density in cell culture processes, due to their ability to track metabolite dynamics in the complex cell culture media of mammalian cells (reviewed in Abu-Absi et al, 2014).…”

High-Throughput Raman Spectroscopy Combined with Innovate Data Analysis Workflow to Enhance Biopharmaceutical Process Development

“…Li at al. utilised Raman spectroscopy for the real-time monitoring of a monoclonal antibody, and were able to distinguish between glycosylated and non-glycosylated molecules [21]. The ability to monitor product quality in real-time opens up significant opportunities for USP optimisation and advanced feedback control solutions.…”

Modeling of a spiral resonant sensor for biological process monitoring

“…The permittivity of a material has been shown to be correlated with indicators of viable cell volume, wet cell weight, cell growth rate, metabolic shifts within the bioreactor, and levels of apoptosis [42][43][44][45][46]. This enables the use of an dielectric probe, which measures the permittivity of a culture, as a PAT tool for monitoring bioprocesses.…”