Increasing the secretory capacity of Saccharomyces cerevisiae for production of single-chain antibody fragments.

Shusta, E V; Raines, R T; Plückthun, A; Wittrup, K D · Nat Biotechnol · 1998

basic_science · Level V

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Abstract

We have produced single-chain antibody fragments (scFv) in Saccharomyces cerevisiae at levels up to 20 mg/L in shake flask culture by a combination of expression level tuning and overexpression of folding assistants. Overexpression of the chaperone BiP or protein disulfide isomerase (PDI) increases secretion titers 2-8 fold for five scFvs. The increases occur for scFv expression levels ranging from low copy to ER-saturating overexpression. The disulfide isomerase activity of PDI, rather than its chaperone activity, is responsible for the secretion increases. A synergistic increase in scFv production occurs upon cooverexpression of BiP and PDI.

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