Harnessing secretory pathway differences between HEK293 and CHO to rescue production of difficult to express proteins.
Bioproduction
CHO
Differential gene expression analysis
HEK293
Protein secretion
Secretory pathway
Transcriptomics
Journal
Metabolic engineering
ISSN: 1096-7184
Titre abrégé: Metab Eng
Pays: Belgium
ID NLM: 9815657
Informations de publication
Date de publication:
07 2022
07 2022
Historique:
received:
23
11
2021
revised:
09
02
2022
accepted:
10
03
2022
pubmed:
19
3
2022
medline:
15
6
2022
entrez:
18
3
2022
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 35301123
pii: S1096-7176(22)00044-1
doi: 10.1016/j.ymben.2022.03.009
pmc: PMC9189052
mid: NIHMS1801357
pii:
doi:
Substances chimiques
Recombinant Proteins
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Research Support, N.I.H., Extramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
171-187Subventions
Organisme : NIGMS NIH HHS
ID : R35 GM119850
Pays : United States
Informations de copyright
Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.
Références
J Biotechnol. 2009 Aug 10;143(1):34-43
pubmed: 19527755
Cytotechnology. 1997;23(1-3):103-11
pubmed: 9094218
Nat Biotechnol. 2013 Aug;31(8):759-65
pubmed: 23873082
J Biotechnol. 2020 Mar 20;312:11-22
pubmed: 32114154
Nat Commun. 2020 Apr 20;11(1):1908
pubmed: 32313013
Sci Rep. 2017 Jan 16;7:40388
pubmed: 28091612
Proc Natl Acad Sci U S A. 2014 Jun 10;111(23):8488-93
pubmed: 24889604
PLoS One. 2016 May 19;11(5):e0155340
pubmed: 27195765
NPJ Syst Biol Appl. 2017 Aug 18;3:22
pubmed: 28845240
Biotechnol Bioeng. 2008 Jun 1;100(2):317-24
pubmed: 18078289
Biotechnol Bioeng. 2016 Sep;113(9):1902-12
pubmed: 26913574
Sci Rep. 2015 Dec 11;5:18016
pubmed: 26657798
Crit Rev Biotechnol. 2018 Sep;38(6):851-867
pubmed: 29262720
Metab Eng. 2014 Jan;21:91-102
pubmed: 23380542
Biotechnol J. 2010 Apr;5(4):393-401
pubmed: 20222103
Biotechnol Bioeng. 2007 May 1;97(1):144-55
pubmed: 17054114
Biotechnol Bioeng. 2003 Nov 5;84(3):292-304
pubmed: 12968283
Bioinformatics. 2014 Aug 1;30(15):2114-20
pubmed: 24695404
Genome Biol. 2014;15(12):550
pubmed: 25516281
N Biotechnol. 2020 Sep 25;58:45-54
pubmed: 32502629
Bioinformatics. 2011 Jun 15;27(12):1739-40
pubmed: 21546393
Cancer Cell. 2013 Sep 9;24(3):289-304
pubmed: 24029229
Methods Mol Med. 2004;94:197-210
pubmed: 14959831
Sci Signal. 2019 Nov 26;12(609):
pubmed: 31772123
Bioinformatics. 2019 Jun 1;35(12):2084-2092
pubmed: 30395178
Mol Biotechnol. 2006 Oct;34(2):225-37
pubmed: 17172668
J Biotechnol. 2012 Oct 31;161(3):336-48
pubmed: 22814405
Biotechnol Adv. 2015 Dec;33(8):1878-96
pubmed: 26523782
Appl Microbiol Biotechnol. 2012 Feb;93(3):917-30
pubmed: 22159888
Biotechnol Bioeng. 2016 Oct;113(10):2241-53
pubmed: 27072894
Nat Biotechnol. 2018 Dec 6;36(12):1136-1145
pubmed: 30520869
Biotechnol Bioeng. 2011 Apr;108(4):977-82
pubmed: 21404269
Mol Cell Proteomics. 2014 Jun;13(6):1611-24
pubmed: 24722731
Biotechnol Prog. 2003 Jan-Feb;19(1):179-84
pubmed: 12573023
Biochemistry. 1981 Nov 10;20(23):6594-9
pubmed: 6796117
Sci Rep. 2020 Nov 4;10(1):18996
pubmed: 33149219
Metab Eng. 2021 Sep;67:237-249
pubmed: 34265400
Biotechnol Adv. 2017 Jan - Feb;35(1):64-76
pubmed: 27931938
F1000Res. 2015 Dec 30;4:1521
pubmed: 26925227
Gene. 2015 Sep 15;569(2):182-90
pubmed: 26026906
BMC Biotechnol. 2006 Dec 22;6:49
pubmed: 17187663
Genome Biol. 2010;11(10):R106
pubmed: 20979621
BMC Biotechnol. 2015 Sep 18;15:87
pubmed: 26382581
Bioinformatics. 2016 Oct 1;32(19):3047-8
pubmed: 27312411
EMBO Rep. 2005 Mar;6(3):239-44
pubmed: 15723043
Front Microbiol. 2017 May 16;8:875
pubmed: 28559891
Nat Commun. 2014 Sep 03;5:4767
pubmed: 25182477
Proc Natl Acad Sci U S A. 2005 Oct 25;102(43):15545-50
pubmed: 16199517
Nat Commun. 2020 Jan 2;11(1):68
pubmed: 31896772
Protein Sci. 2014 May;23(5):517-25
pubmed: 24510886
Cell Syst. 2021 Sep 22;12(9):873-884.e4
pubmed: 34171228
Cytotechnology. 2013 Dec;65(6):993-1002
pubmed: 24026344
Metab Eng. 2019 Jul;54:54-68
pubmed: 30851381
Nat Rev Mol Cell Biol. 2013 Oct;14(10):630-42
pubmed: 24026055
Nature. 1997 Jun 5;387(6633):607-11
pubmed: 9177349
Biotechnol Bioeng. 2003 Feb 20;81(4):496-504
pubmed: 12491534
Biotechnol J. 2018 Mar;13(3):e1700231
pubmed: 29316330
Biotechnol Bioeng. 2018 Aug;115(8):2087-2100
pubmed: 29704459
Biotechnol Prog. 2011 Jan-Feb;27(1):220-31
pubmed: 21312369
Mol Cell. 2019 Jul 25;75(2):394-407.e5
pubmed: 31227230
Clin Immunol. 2006 May;119(2):135-45
pubmed: 16458072
Crit Rev Biotechnol. 2016 Dec;36(6):1110-1122
pubmed: 26383226
Sci Rep. 2016 Aug 11;6:31365
pubmed: 27509843
Cytotechnology. 2015 Mar;67(2):237-54
pubmed: 24504562
Nucleic Acids Res. 2000 Jan 1;28(1):27-30
pubmed: 10592173
Nat Methods. 2017 Apr;14(4):417-419
pubmed: 28263959
J Virol. 2005 Dec;79(24):15277-88
pubmed: 16306599
Biotechnol Bioeng. 2018 Jan;115(1):165-173
pubmed: 28921524
BMC Biotechnol. 2011 Jul 24;11:78
pubmed: 21781345
Biotechnol Lett. 2014 Aug;36(8):1581-93
pubmed: 24752815