Development of a versatile high-temperature short-time (HTST) pasteurization device for small-scale processing of cell culture medium formulations

Abstract

The compatibility of CHO cell culture medium formulations with all stages of the bioprocess must be evaluated through small-scale studies prior to scale-up for commercial manufacturing operations. Here, we describe the development of a bespoke small-scale device for assessing the compatibility of culture media with a widely implemented upstream viral clearance strategy, high-temperature short-time (HTST) treatment. The thermal stability of undefined medium formulations supplemented with soy hydrolysates was evaluated upon variations in critical HTST processing parameters, namely, holding times and temperatures. Prolonged holding times of 43 s at temperatures of 110 °C did not adversely impact medium quality while significant degradation was observed upon treatment at elevated temperatures (200 °C) for shorter time periods (11 s). The performance of the device was benchmarked against a commercially available mini-pilot HTST system upon treatment of identical formulations on both platforms. Processed medium samples were analyzed by untargeted LC-MS/MS for compositional profiling followed by chemometric evaluation, which confirmed the observed degradation effects caused by elevated holding temperatures but revealed comparable performance of our developed device with the commercial mini-pilot setup. The developed device can assist medium optimization activities by reducing volume requirements relative to commercially available mini-pilot instrumentation and by facilitating fast throughput evaluation of heat-induced effects on multiple medium lots.

Interaction of cell culture process parameters for modulating mAb afucosylation

“…Stability issues with cell culture media have been previously associated with HTST treatment (Floris et al, 2018) or with extended storage in the cold or at room temperature (Hossler, McDermott, Racicot, & Fann, 2013;Kuschelewski, Schnellbaecher, Pering, Wehsling, & Zimmer, 2017;McCoy, Costa, & Morris, 2015). These extensive investigations identified a number of root causes for media F I G U R E 3 (a) Effect of pCO 2 model, media hold, and Mn supplementation on afucosylation of mAb at time of harvest (Day 12) in a full-factorial DOE study.…”

“…The first process parameter selected for investigation as an interaction partner for pCO 2 was manganese (Mn) because it is a cofactor for multiple glycosylation enzymes (Rouiller et al, 2014) and it has been used to modulate glycosylation levels in CHO cell culture studies (Gramer et al, 2011;Surve & Gadgil, 2014). The final process parameter selected for investigation was high-temperature short-time (HTST) treatment of culture media because this operation may be implemented in commercial manufacturing facilities as an additional safety barrier against adventitious agents (Floris, Curtin, Kaisermayer, Lindeberg, & Bones, 2018;Pohlscheidt et al, 2014); by contrast, small-scale operations typically use media without HTST treatment. The next process parameter selected for investigation was media hold duration (at 37°C) because its impact on product quality attributes has not been previously reported.…”

“…In large-scale operations, culture media are typically warmed and then held at the cultivation temperature in bioreactors before inoculation; by contrast, small-scale operations typically do not involve a media hold step because the media are added at the time of bioreactor inoculation. The final process parameter selected for investigation was high-temperature short-time (HTST) treatment of culture media because this operation may be implemented in commercial manufacturing facilities as an additional safety barrier against adventitious agents (Floris, Curtin, Kaisermayer, Lindeberg, & Bones, 2018;Pohlscheidt et al, 2014); by contrast, small-scale operations typically use media without HTST treatment.…”

Review: High temperature short time treatment of cell culture media and feed solutions to mitigate adventitious viral contamination in the biopharmaceutical industry

“…35 Among them, some are known to be owned by Amgen, and Biomarin. 26,29,33,34,36 Eli Lilly also showed interest in this technology with a study at small-scale. 24 The Life Science business of Merck KGaA offers the opportunity for outsourcing HTST treatment of high-risk raw materials such as glucose at their two redundant large-scale production sites.…”

Consequences of trace metal variability and supplementation on Chinese hamster ovary (CHO) cell culture performance: A review of key mechanisms and considerations

“…After dissociating in solution, these ions can form small concentrations of ionic phosphates (Cao, Stimpfl, Wen, Frank & Hunter, ). During HTST treatments, elevated temperatures in excess of 100°C can drive these ionic phosphates and other inorganic salts out of solution (Cao, Loussaert & Wen, ; Cao et al, ; Floris, Curtin, Kaisermayer, Lindeberg & Bones, ; Pohlscheidt et al, ). Consequently, lot‐to‐lot variabilities of trace metal content are accentuated after the precipitating solids are removed from the solution.…”