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 (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.
Evans
Leslie
L
Novozymes Biopharma UK Ltd., Castle Court, 59 Castle Boulevard, Nottingham NG7 1FD, United Kingdom. lsre@novozymes.com
Hughes
Miranda
M
Waters
Joanne
J
Cameron
Jason
J
Dodsworth
Neil
N
Tooth
David
D
Greenfield
Anthony
A
Sleep
Darrell
D
D000418:Albumins / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D019540:Area Under Curve; D019677:Artificial Gene Fusion; D019149:Bioreactors; D005285:Fermentation / Q000235:genetics; D016650:Fluorescein-5-isothiocyanate / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D006207:Half-Life; D006801:Humans; D008657:Metabolic Clearance Rate; D051381:Rats; D017208:Rats, Wistar; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification / Q000493:pharmacokinetics; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D012709:Serum Albumin / Q000235:genetics* / Q000378:metabolism; D057127:Single-Chain Antibodies / Q000235:genetics / Q000276:immunology*
D000418:Albumins; D005456:Fluorescent Dyes; D011993:Recombinant Fusion Proteins; D012709:Serum Albumin; D057127:Single-Chain Antibodies; D016650:Fluorescein-5-isothiocyanate