Bax and Bak knockout apoptosis‐resistant Chinese hamster ovary cell lines significantly improve culture viability and titer in intensified fed‐batch culture process

Danming Tang; Cynthia Lam; Niels BauerSimon AuslaenderBrad SnedecorMichael W. LairdShahram Misaghi

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Bax and Bak knockout apoptosis‐resistant Chinese hamster ovary cell lines significantly improve culture viability and titer in intensified fed‐batch culture process

机译：Bax and Bak knockout apoptosis‐resistant Chinese hamster ovary cell lines significantly improve culture viability and titer in intensified fed‐batch culture process

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Abstract In the field of therapeutic protein production, process intensification strategies entailing higher starting cell seeding densities, can potentially increase culture productivity, lower cost of goods and improve facility utilization. However, increased cell densities often trigger apoptotic cell death at the end of the cell culture process and thus reduce total viable cell count. Apoptosis‐resistant Chinese hamster ovary cell lines may offer the possibility to diminish this undesired outcome of the intensified production process. In this study, we have generated and tested Bax/Bak double‐knock‐out (DKO) apoptosis resistant hosts to express standard and bispecific antibodies, as well as complex molecules in intensified production processes both as pools and single cell clones, and at different scales. In all cases, therapeutic proteins expressed from clones or pools generated from the Bax/Bak DKO hosts showed not only better viability but also enabled extended productivity in the later stages of the 14‐day intensified production process. The product qualities of the produced molecules were comparable between Bax/Bak DKO and wild type cells. Overall, we showed that Bax/Bak DKO apoptosis‐resistant host cell lines significantly improve viability and volumetric productivity of the intensified production cultures without altering product qualities.

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《Biotechnology Progress》 |2022年第2期|3228-1-3228-9|共9页
作者
Danming Tang; Cynthia Lam; Niels BauerSimon AuslaenderBrad SnedecorMichael W. LairdShahram Misaghi;
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作者单位

Genentech Inc.;

Roche Pharma Research and Early Development (pRED), Roche Innovation Center Munich;

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原文格式 PDF
正文语种英语
中图分类生物科学;
关键词
apoptosis‐resistant; Bak; Bax; Chinese hamster ovary; CRISPR; fed‐batch; gene knockout; process intensification;

Bax and Bak knockout apoptosis‐resistant Chinese hamster ovary cell lines significantly improve culture viability and titer in intensified fed‐batch culture process

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