Production of polypeptides

C - Chemistry – Metallurgy – 12 – P

Patent

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Details

C12P 21/02 (2006.01) C07K 16/00 (2006.01) C07K 16/22 (2006.01) C07K 16/30 (2006.01) C12N 5/00 (2006.01)

Patent

CA 2578141

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

L'invention porte sur un système amélioré de production à grande échelle de protéines et/ou de polypeptides dans une culture cellulaire, notamment dans des milieux caractérisés par au moins: i) une concentration d'acides aminés cumulés supérieure à environ 70 mM; ii) un rapport molaire de glutamine cumulée/asparagine cumulée inférieur à environ 2; iii) un rapport molaire de glutamine cumulée/acides aminés cumulés inférieur à environ 0.2; iv) un rapport molaire d'ions inorganiques cumulés/acides aminés cumulés compris entre environ 0,4 et 1; ou v) une concentration combinée de glutamine cumulée et d'asparagine cumulée comprise entre environ 16 et 36 mM. L'utilisation de ce système permet de parvenir à des niveaux élevés de production de protéines et de réduire l'accumulation de certains facteurs indésirables tels que l'ammonium et/ou le lactate. De plus, les procédés de mise en culture présentent un écart de température tel que, généralement, une baisse de la température lorsque la culture a atteint environ 20 à 80 % de sa densité cellulaire maximale. En variante ou de plus, cette invention porte sur des procédés dans lesquels, après avoir atteint une crête, les taux de lactate et/ou d'ammonium de la culture diminuent au fil du temps.

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