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Secretory pathway differences between HEK293 and CHO enable rescue of difficult to express human proteins

ABSTRACT: Secretory pathway differences between HEK293 and CHO enable rescue of difficult to express human proteins

PROVIDER: PRJNA662582 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR12618000.fastq.gz	Fastqsanger.gz
	SRR12618100.fastq.gz	Fastqsanger.gz
	SRR12618001.fastq.gz	Fastqsanger.gz
	SRR12618101.fastq.gz	Fastqsanger.gz
	SRR12618002.fastq.gz	Fastqsanger.gz

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Secretory pathway differences between HEK293 and CHO enable rescue of difficult to express human proteins

Project description:Secretory pathway differences between HEK293 and CHO enable rescue of difficult to express human proteins

| PRJNA662582 | ENA

Secretory pathway differences between HEK293 and CHO enable rescue of difficult to express human proteins

Project description:Biologics have emerged as leading blockbuster therapeutics, but many advanced recombinant protein moieties remain difficult to produce. Here we identify bottlenecks limiting expression of challenging recombinant human proteins through a systems biology analysis of the transcriptomes of CHO and HEK293. Surprisingly, one third of the challenging human proteins displayed an improved secretion upon host cell swapping from CHO to HEK293. While most components of the secretory machinery showed comparable expression levels in both expression hosts, a key set of genes with extreme expression variation was identified. Among these, ATF4 and SRP9 were validated as general productivity boosters in CHO. Further, we found that more heavily glycosylated products benefit more from the elevated activities of the N- and O-glycosyltransferases found in HEK293 cells. Collectively, our results demonstrate the utilization of HEK293 cells for expression rescue of human proteins and suggest a strategy for identification of secretory pathway components improving yield and quality in HEK293 and CHO.

2022-06-22 | GSE157729 | GEO

Harnessing secretory pathway differences between HEK293 and CHO to rescue production of difficult to express proteins.

Project description: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.

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CHO-S transcriptome in batch culture

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Cricetulus griseus strain:CHO

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