Enhanced protein expression through strain selection, gene disruption, improved vector design and co-expression of endogenous chaperones
© Sleep et al; licensee BioMed Central Ltd. 2006
Published: 10 October 2006
The use of Saccharomyces cerevisiae as a host system has been limited by the perception of limited secretion capacity, unstable episomal vectors and aberrant glycosylation. Solutions to all of these limitations are now available.
The expression vectors have been further enhanced to facilitate the stable co-expression of multiple proteins. When one of these proteins is a chaperone, the titre of co-expressed recombinant transferrin was increased 15-fold. The applicability of this system has been demonstrated with a wide range of heterologous proteins and is scalable from 10 mL shake flask to cGMP manufacture at high cell density fermentation (8,000 L) in a defined synthetic medium; designed to be integrated with cost-efficient downstream processing.
Significant intra-strain variability and unstable episomal plasmid systems have limited the usefulness of Saccharomyces cerevisiae as an industrial host for the production of biopharmaceuticals. However co-enhancement of the episomal vector system and the host strains is not only possible but has led to significant improvements in recombinant protein production.
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