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Xiang, Shaoxun; Zhang, Jinliang; Yu, Le; Tian, Jun; Tang, Wenxiu; Tang, Hao; Xu, Kecui; Wang, Xin; Cui, Yanyan; Ren, Kaidi; Cao, Weijia; Su, Yuning; Zhou, Weichang
Biotechnology and bioengineering, February 2024, 2024-Feb, 2024-02-00, 20240201, Volume: 121, Issue: 2Journal Article
Intensified fed‐batch (IFB), a popular cell culture intensification strategy, has been widely used for productivity improvement through high density inoculation followed by fed‐batch cultivation. However, such an intensification strategy may counterproductively induce rapidly progressing cell apoptosis and difficult‐to‐sustain productivity. To improve culture performance, we developed a novel cell culture process intermittent‐perfusion fed‐batch (IPFB) which incorporates one single or multiple cycles of intermittent perfusion during an IFB process for better sustained cellular and metabolic behaviors and notably improved productivity. Unlike continuous perfusion or other semi‐continuous processes such as hybrid perfusion fed‐batch with only early‐stage perfusion, IPFB applies limited times of intermittent perfusion in the mid‐to‐late stage of production and still inherits bolus feedings on nonperfusion days as in a fed‐batch culture. Compared to IFB, an average titer increase of ~45% was obtained in eight recombinant CHO cell lines studied. Beyond IPFB, ultra‐intensified IPFB (UI‐IPFB) was designed with a markedly elevated seeding density of 20–80 × 106 cell/mL, achieved through the conventional alternating tangential flow filtration (ATF) perfusion expansion followed with a cell culture concentration step using the same ATF system. With UI‐IPFB, up to ~6 folds of traditional fed‐batch and ~3 folds of IFB productivity were achieved. Furthermore, the application grounded in these two novel processes showed broad‐based feasibility in multiple cell lines and products of interest, and was proven to be effective in cost of goods reduction and readily scalable to a larger scale in existing facilities.
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