Cell culture process

Details Cell culture process Cell culture process

C - Chemistry, Metallurgy

12

N

C12N 15/58 (2006.01) C12N 9/72 (2006.01) C12P 21/00 (2006.01)

Patent

CA 2368797

A glycoprotein is produced by a process comprising culturing mammalian host cells expressing nucleic acid encoding said glycoprotein in the presence of (a) a factor that modifies growth state in a cell culture, (b) a divalent metal cation that can adopt and prefers and octahedral coordination geometry, and/or (c) a plasma component. In this process, the occupancy of an N-linked glycosylation site occupied only in a fraction of a glycoprotein is enhanced. Such culturing is preferably carried out at a temperature of between about 30 ~C and 35 ~C and/or in the presence of up to about 2mM of a butyrate salt and/or in the presence of a cell-cycle inhibitor.

L'invention concerne une glycoprotéine obtenue par la mise en oeuvre d'un procédé consistant à cultiver des cellules hôtes de mammifère exprimant un acide nucléique codant pour cette glycoprotéine en présence de a) un facteur modifiant l'état de croissance d'une culture cellulaire, b) un cation métallique divalent adoptant de préférence une configuration octaèdre et/ou c) un composant de plasma. Selon ce procédé, l'occupation d'un site de glcosylation à liaison N occupé uniquement dans un fragment d'une glycoprotéine est améliorée. Cette culture est soumise de préférence à une température allant d'environ 30 a 35 ·C et/ou en présence de jusqu'à environ 2 mM d'un sel de butyrate et/ou en présence d'un inhibiteur du cycle cellulaire.

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