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

Protein Expr Purif. 2010 Oct;73(2):113-24. doi: 10.1016/j.pep.2010.05.009. Epub 2010 May 27.

Abstract

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.

MeSH terms

  • Albumins / genetics
  • Albumins / metabolism*
  • Animals
  • Area Under Curve
  • Artificial Gene Fusion*
  • Bioreactors
  • Fermentation / genetics
  • Fluorescein-5-isothiocyanate / metabolism
  • Fluorescent Dyes / metabolism
  • Half-Life
  • 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 / genetics
  • Saccharomyces cerevisiae / immunology
  • Saccharomyces cerevisiae / metabolism*
  • Serum Albumin / genetics*
  • Serum Albumin / metabolism
  • Single-Chain Antibodies / genetics
  • Single-Chain Antibodies / immunology*

Substances

  • Albumins
  • Fluorescent Dyes
  • Recombinant Fusion Proteins
  • Serum Albumin
  • Single-Chain Antibodies
  • Fluorescein-5-isothiocyanate