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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: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.

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