Overexpression of desired proteins in eukaryotic cells...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/67 (2006.01) C07K 14/47 (2006.01) C07K 14/475 (2006.01) C12N 5/10 (2006.01)

Patent

CA 2338820

The invention concerns eukaryotic cells useful in protein expression comprising (a) an inserted nucleic acid encoding a cyclin D gene product and (b) an inserted nucleic acid encoding a protein of interest, wherein the cyclin D gene product and the protein of interest are expressed in the cell. The invention also concerns a process for producing a protein of interest, which comprises (a) inserting into a eukaryotic cell a nucleic acid encoding a cyclin D gene product and a nucleic acid encoding a protein of interest; (b) culturing the cell under conditions permitting the expression of the protein of interest; and (c) isolating the protein of interest. The cells are preferably mammalian, with CHO cells most preferred. The cyclin D gene product is preferably of human origin. Suitable proteins of interest include erythropoietin (EPO), osteoprotegerin (OPG), OPG-Fc, leptin, Fc-leptin, and Novel Erythropoiesis Stimulating Protein (NESP).

La présente invention concerne des cellules eucaryotes utilisées dans l'expression protéique comprenant à: (a) un acide nucléique introduit codant un produit génique de la cycline D et (b) un acide nucléique introduit codant la protéine recherchée, le produit génique de la cycline D et la protéine recherchée étant exprimés dans la cellule. Par ailleurs, cette invention concerne un procédé de production d'une protéine recherchée consistant à: (a) introduire dans une cellule eucaryote un acide nucléique codant un produit génique de la cycline D et un acide nucléique codant une protéine recherchée; (b) mettre en culture la cellule en question dans des conditions permettant l'expression de la protéine recherchée; et enfin, (c) isoler la protéine recherchée. Ces cellules sont de préférence des cellules mammifères, et plus préférablement des cellules CHO. Le produit génique de la cycline D est préférablement d'origine animale. Les protéines appropriées comprennent l'érythropoïétine (EPO), l'ostéoprotégérine (OPG), OPG-FC, la leptine, la FC-leptine, ainsi que la nouvelle protéine de stimulation de l'érythropoïèse (NESP).

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