Wyeth

Wyeth Research Ireland Limited

Genetics Institute, Inc.

Pfizer Corporation

Pfizer Ireland Pharmaceuticals

Denis Drapeau

Daniel T Lasko

Yen-Tung Luan

Wenge Wang

Daniel R Lasko

James R Mercer

S Robert Adamson

Gregory Walter Hiller

Ryan Nolan

Jose Manuel Gomes

Yen Tung Luan

Terry Cardoza Stanek

Douglas A Miller

Paul Thoday

Terry Cardoza

Gregg Nyberg

Jessica Snow

Judy Chou

Ryan P Nolan

An improved system for large scale production of polypeptides in cell culture is provided. In accordance with the present invention, cells expressing a polypeptide of interest are grown in media that contain copper, glutamate or both. The use of such a system allows production of polypeptides in which misfolding and/or aggregation are reduced, and in which total glycosylation is increased. Polypeptides expressed in accordance with the present invention may be advantageously used in the preparation of pharmaceutical, agricultural or other commercial compositions.

Use of copper and glutamate in cell culture for production of polypeptides

This invention relates to methods for rationally designing cell culture media for use in cell cultures, e.g., cell cultures employed in polypeptide production; cell culture media designed with the disclosed methods; methods of producing a polypeptide of interest, e.g., an antibody, using such media; polypeptides produced using the methods and media disclosed herein; and pharmaceuticals compositions containing such polypeptides. The rationally designed media contain a concentration of an amino acid that is calculated for use in cell mass, a concentration of the amino acid that is calculated for use in cell maintenance, and a concentration of the amino acid that is calculated for incorporation into the polypeptide of interest.

Rationally designed media for cell culture

Methods and processes for improved recombinant protein production are provided. The methods are useful for production of growth factors, particularly those of the TGF-&#x3b2; superfamily, including bone morphogenetic proteins (BMPs), such as BMP-2. Suitable host cells are cultured in media where iron is present at a concentration of at least 2.25 &#x3bc;M and if pyridoxal is present, it makes up less than 55% of the molar concentration of vitamin B6 in the media.

Methods for enhanced production of bone morphogenetic proteins

Methods of improving protein production in animal cell cultures are provided. Cell culture methods are presented wherein glucose is fed in a restricted manner to cell culture; this restricted feeding of glucose to the cell culture results in lactate production being controlled to a low level. The restricted feeding of glucose in a fed-batch process is not accomplished through a constant-rate feeding of glucose, and the restricted feeding need not depend on sampling. Instead, restricted feeding of glucose to the culture is accomplished through feeding of glucose to the culture at a rate that is a function of an expected or a premodeled rate of glucose consumption by the animal cells when exposed to medium containing a high level of glucose. Because lactate production is controlled to low levels, recombinant protein production is increased.

Restricted glucose feed for animal cell culture

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

Cell culture media are provided containing high L-cystine concentration and low L-glutamic acid concentration. The media are useful for recombinant production of proteins using mammalian cell cultures.

Medium and methods for culturing mammalian cho cells

Methods and nutrient media are disclosed useful for adapting mammalian cell lines to culture at increased cell densities.

Adaption of mammalian cell lines to high cell densities

Growing community of inventors

Salem, NH, United States of America

Denis Drapeau