Low Shear Stress Increases Recombinant Protein Production and High Shear Stress Increases Apoptosis in Human Cells

Caijuan Zhan; Gholamreza Bidkhori; Hubert Schwarz; Magdalena Malm; Aman Mebrahtu; Ray Field; Christopher Sellick; Diane Hatton; Paul Varley; Adil Mardinoglu; Johan Rockberg; Veronique Chotteau

doi:10.1016/j.isci.2020.101653

Low Shear Stress Increases Recombinant Protein Production and High Shear Stress Increases Apoptosis in Human Cells

iScience. 2020 Oct 7;23(11):101653. doi: 10.1016/j.isci.2020.101653. eCollection 2020 Nov 20.

Authors

Caijuan Zhan^{1

2

3}, Gholamreza Bidkhori⁴, Hubert Schwarz^{1

2

3}, Magdalena Malm^{5

2}, Aman Mebrahtu^{5

2}, Ray Field⁶, Christopher Sellick⁶, Diane Hatton⁶, Paul Varley⁶, Adil Mardinoglu⁴, Johan Rockberg^{5

2

3}, Veronique Chotteau^{1

2

3}

Affiliations

¹ KTH - Cell Technology Group (CETEG), Department of Industrial Biotechnology, 106 91, Stockholm, Sweden.
² Wallenberg Centre for Protein Research (WCPR), 106 91 Stockholm, Sweden.
³ AdBIOPRO, Competence Centre for Advanced Bioproduction by Continuous Processing, Stockholm, Sweden.
⁴ Science for Life Laboratory, KTH - Royal Institute of Technology, 171 21, Stockholm, Sweden.
⁵ KTH - Royal Institute of Technology, Department of Protein Science, 106 91 Stockholm, Sweden.
⁶ BioPharmaceutical Development, AstraZeneca, Cambridge, UK.

Abstract

Human embryonic kidney cells HEK293 can be used for the production of therapeutic glycoproteins requiring human post-translational modifications. High cell density perfusion processes are advantageous for such production but are challenging due to the shear sensitivity of HEK293 cells. To understand the impact of hollow filter cell separation devices, cells were cultured in bioreactors operated with tangential flow filtration (TFF) or alternating tangential flow filtration (ATF) at various flow rates. The average theoretical velocity profile in these devices showed a lower shear stress for ATF by a factor 0.637 compared to TFF. This was experimentally validated and, furthermore, transcriptomic evaluation provided insights into the underlying cellular processes. High shear caused cellular stress leading to apoptosis by three pathways, i.e. endoplasmic reticulum stress, cytoskeleton reorganization, and extrinsic signaling pathways. Positive effects of mild shear stress were observed, with increased recombinant erythropoietin production and increased gene expression associated with transcription and protein phosphorylation.

Keywords: Bioengineering; Biophysics; Cell Biology.