Shift to high‐intensity, low‐volume perfusion cell culture enabling a continuous, integrated bioprocess

Abstract

In order to address the increasing demand for biologics, cell culture intensification using perfusion offers significantly higher productivities while also reducing manufacturing costs, especially when part of an integrated, continuous bioprocess. An initial study of a long‐duration perfusion process using a cell‐bleed to maintain a target cell density observed a 2.1‐fold higher cell‐specific productivity and a gradual decline in the culture growth rate when perfused at an overall lower rate. Subsequent studies sought an alternative process that largely reduced the overall volume of media needed by first perfusing at a high cell‐specific perfusion rate (CSPR) to support a high cell density followed by continued perfusion at a low CSPR to promote a more productive stationary phase. This high intensity, low‐volume perfusion (HILVOP) process achieved cumulative volumetric productivities of 1.5–1.6 g/L/day with two CHO cell lines. When compared to each cell line's respective commercial‐ready, fed‐batch process, a 3.1–3.8‐fold productivity increase was demonstrated while yielding similar product quality. Furthermore, the higher productivity achieved with HILVOP used 6.6–12.3‐fold less media than a similarly productive long‐duration process. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:1472–1481, 2018

Rapid development of clone‐specific, high‐performing perfusion media from established feed supplements

“…Such attempts to decouple productivity from growth were already proven by media additives such as small molecule cell growth inhibitors, or altered sodium–potassium levels to arrest cells in the G0/G1 phase . Alternatively, process control strategies were used to keep cells in a physiological state using lactate consumption and the resulting rise in culture pH to regulate perfusion media addition or minimizing growth and maximize productivity by reduced culture temperatures or limiting CSPR . Culture pH, pCO 2 levels, and osmolality were also shown to profoundly impact cell growth and productivity and may be used for future perfusion control strategies.…”

Wide‐surface pore microfiltration membrane drastically improves sieving decay in TFF‐based perfusion cell culture and streamline chromatography integration for continuous bioprocessing

“…Emerging capacity shortage and increased demand for cost‐appropriate therapeutic proteins is prompting a shift in biomanufacturing. Its conventional rigid stainless‐steel structure is being replaced with state‐of‐the‐art approaches, leveraging single‐use technologies as well as continuous and connected bioprocesses (Gagnon, Nagre, Wang, & Hiller, 2018; Klutz et al, 2015; Zydney, 2016). These more modern processes offer higher productivities, reduced facility footprint, and increased operational flexibility compared to the gold standard fed‐batch processes (Fisher et al, 2019 ) .…”

Hydrocyclones as cell retention device for CHO perfusion processes in single‐use bioreactors

“…In past decades, perfusion had remained almost completely confined to the production of unstable proteins, and this was usually attributed to the higher technical complexity, the limitations of the cell retention devices then available, and the more dilute harvest due to continuous medium exchange. The advances in single‐use technology, the development of more appropriate cell retention devices, and the progress in culture media and feeding strategies (Gagnon, Nagre, Wang, & Hiller, 2018; Konstantinov et al, 2006; Sieck et al, 2017), which allowed cell densities in the order of 10 8 cells per milliliter (Clincke et al, 2013) and high product titers to be attained, were decisive in turning perfusion into the current mode of operation of choice, also for stable proteins.…”