Zymogenetics, Inc.

Bristol-myers Squibb Compnay

Bruce L Zamost

Robert M Dedinsky

Hong Yan Liu

Karen S De Jongh

Douglas C Covert

Chung Chan

Susan D Holderman

Jeffrey D Meyer

Geoffrey F Lee

The expression vectors and methods using anexpression system for the large scale production of IL-29 are described. The vectors utilize the IL-29 coding sequence with specific changes in nucleotides in order to optimize codons and mRNA secondary structure for translation in. Also included are methods of producing, purifying and pegylating an IL-29 polypeptide.

Production and purification of IL-29

The expression vectors and methods using anexpression system for the large scale production of IL-21 are described. The vectors utilize the IL-21 coding sequence with specific changes in nucleotides in order to optimize codons and mRNA secondary structure for translation in. Using the expression vectors, the IL-21 gene was produced into a level of greater than 1 g/L in fed batch fermentation. Also included are OmpT deficientstrains transformed with an IL-21 expression vector.

IL-21 production in prokaryotic hosts

The expression vectors and methods using anexpression system for the large scale production of IL-21 are described. The vectors utilize the IL-21 coding sequence with specific changes in nucleotides in order to optimize codons and mRNA secondary structure for translation inUsing the expression vectors, the IL-21 gene was produced into a level of greater than 1 g/L in fed batch fermentation. Also included are OmpT deficientstrains transformed with an IL-21 expression vector.

A yeast fermentation medium that does not contain yeast cell extract but is comprised of a mixture of oleic acid, lactic acid and palmitic acid. Using this fermentation medium yields of factor XIII were increased 250%.

Fermentation medium and method

Methylotrophic yeasts are useful hosts for the production of commercially valuable recombinant proteins. However, the development of large-scale cultures of recombinant methylotrophic yeasts has been hindered by the formation of precipitation in culture media. A new soluble minimal medium overcomes this problem. Moreover, new feeding schemes provide cultures of high biomass, which produce biologically active recombinant protein.

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Seattle, WA, United States of America

Bruce L Zamost