1. Harnessing secretory pathway differences between HEK293 and CHO to rescue production of difficult to express proteins
- Author
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M. Moradi Barzadd, Anna-Luisa Volk, Diane Hatton, Raymond Field, Aman Mebrahtu, Paul Varley, R. G. Roth, Veronique Chotteau, N. Wistabacka, Fredrik Edfors, Chih-Chung Kuo, Magdalena Malm, Nathan E. Lewis, Ronia Razavi, T. Graslund, Johan Rockberg, David Kotol, and M. Lunqvist
- Subjects
Secretory Pathway ,HEK 293 cells ,Bioengineering ,PDIA3 ,CHO Cells ,Biology ,Applied Microbiology and Biotechnology ,Recombinant Proteins ,Cell biology ,law.invention ,Transcriptome ,Cricetulus ,HEK293 Cells ,law ,Cricetinae ,Recombinant DNA ,Animals ,Humans ,Secretion ,HSPA8 ,Gene ,Secretory pathway ,Biotechnology - Abstract
Biologics represent the fastest growing group of therapeutics, but many advanced recombinant protein moieties remain difficult to produce. Here, we identify metabolic engineering targets limiting expression of recombinant human proteins through a systems biology analysis of the transcriptomes of CHO and HEK293 during recombinant expression. In an expression comparison of 24 difficult to express proteins, one third of the challenging human proteins displayed improved secretion upon host cell swapping from CHO to HEK293. Guided by a comprehensive transcriptomics comparison between cell lines, especially highlighting differences in secretory pathway utilization, a co-expression screening of 21 secretory pathway components validated ATF4, SRP9, JUN, PDIA3 and HSPA8 as productivity boosters in CHO. Moreover, more heavily glycosylated products benefitted more from the elevated activities of the N- and O-glycosyltransferases found in HEK293. Collectively, our results demonstrate the utilization of HEK293 for expression rescue of human proteins and suggest a methodology for identification of secretory pathway components for metabolic engineering of HEK293 and CHO.
- Published
- 2022