The production, characterisation and enhanced pharmacokinetics of scFv–albumin fusions expressed in Saccharomyces cerevisiae

An expression system is described for the production of monomeric scFvs and scFv antibody fragments genetically fused to human albumin (at either the N- or C-terminus or both). Based upon strains of Saccharomyces cerevisiae originally developed for the production of a recombinant human albumin (Reco...

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Bibliographic Details
Published in:Protein expression and purification 2010-10, Vol.73 (2), p.113-124
Main Authors: Evans, Leslie, Hughes, Miranda, Waters, Joanne, Cameron, Jason, Dodsworth, Neil, Tooth, David, Greenfield, Anthony, Sleep, Darrell
Format: Article
Language:English
Subjects:
Albumin fusions
Albumins - genetics
Albumins - metabolism
Animals
Area Under Curve
Artificial Gene Fusion
Bioreactors
Fermentation - genetics
Fluorescein-5-isothiocyanate - metabolism
Fluorescent Dyes - metabolism
Half-Life
Heterologous protein expression
Humans
Metabolic Clearance Rate
Rats
Rats, Wistar
Recombinant Fusion Proteins - biosynthesis
Recombinant Fusion Proteins - immunology
Recombinant Fusion Proteins - isolation & purification
Recombinant Fusion Proteins - pharmacokinetics
Saccharomyces cerevisiae
Saccharomyces cerevisiae - genetics
Saccharomyces cerevisiae - immunology
Saccharomyces cerevisiae - metabolism
scFv
scFv–albumin fusions
Serum Albumin - genetics
Serum Albumin - metabolism
Single-Chain Antibodies - genetics
Single-Chain Antibodies - immunology
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Description
Summary:An expression system is described for the production of monomeric scFvs and scFv antibody fragments genetically fused to human albumin (at either the N- or C-terminus or both). Based upon strains of Saccharomyces cerevisiae originally developed for the production of a recombinant human albumin (Recombumin®) this system has delivered high levels of secreted product into the supernatant of shake flask and high cell density fed-batch fermentations. Specific binding to the corresponding ligand was demonstrated for each of the scFvs and scFv–albumin fusions and pharmacokinetic studies showed that the fusion products had greatly extended circulatory half-lives. The system described provides an attractive alternative to other microbial systems for the manufacture of this type of product.
ISSN:1046-5928
1096-0279
DOI:10.1016/j.pep.2010.05.009