Wyeth Research Ireland Limited

Pfizer Corporation

Wyeth

Pfizer Ireland Pharmaceuticals

Daniel R Lasko

Wenge Wang

Denis Drapeau

Yen-Tung Luan

James R Mercer

Jason Arnold Lotvin

Michael Allen Hanson

Weiqiang Sun

Scott Ellis Lomberk

Jonathan Patrick Kinross

Sujata Kaushikbhai Patel-Brown

Peter Anthony Tomasello

Stephan M Koza

Sunil Gururao Desai

The present invention relates to media and fermentation methods for producing polysaccharides in bacterial cell culture. In one aspect, the invention relates to a complex culture medium comprising a vegetable hydrolysate, a yeast extract, and a carbon source. In another aspect, the invention relates to a defined media having a total amino acid concentration greater than about 50 mM. A further aspect of the invention relates to the use of fed batch and perfusion fermentation methods for cultivating polysaccharide-producing bacteria.

Media and fermentation methods for producing polysaccharides in bacterial cell culture

An improved system for large scale production of glycoproteins in cell culture is provided. In accordance with the present invention, cells expressing a glycoprotein are grown in media that contain manganese at a concentration of between approximately 10 and 600 nM. The use of such a system allows production of a glycoprotein with an increased glycosylation pattern and/or a glycosylation pattern that more accurately reflects the glycosylation pattern of the naturally occurring glycoprotein. A glycoprotein expressed in accordance with the present invention may be advantageously used in the preparation of pharmaceutical compositions.

Production of glycoproteins

An improved system for large scale production of proteins and/or polypeptides in cell culture, particularly in media characterized by one or more of: i) a cumulative amino acid concentration greater than about 70 mM; ii) a molar cumulative glutamine to cumulative asparagine ratio of less than about 2; iii) a molar cumulative glutamine to cumulative total amino acid ratio of less than about 0.2; iv) a molar cumulative inorganic ion to cumulative total amino acid ratio between about 0.4 to 1; or v) a combined cumulative glutamine and cumulative asparagine concentration between about 16 and 36 mM, is provided. The use of such a system allows high levels of protein production and lessens accumulation of certain undesirable factors such as ammonium and/or lactate. Additionally, culture methods including a temperature shift, typically including a decrease in temperature when the culture has reached about 20-80% of it maximal cell density, are provided. Alternatively or additionally, the present invention provides methods such that, after reaching a peak, lactate and/or ammonium levels in the culture decrease over time.

Production of anti-abeta

Production of TNFR-Ig

Production of polypeptides

Growing community of inventors

Medford, MA, United States of America

Daniel R Lasko