Antibody composition-producing cell
View Patent ↗The present invention relates to a cell for the production of an antibody molecule such as an antibody useful for various diseases having high antibody-dependent cell-modulated cytotoxic activity, a fragment of the antibody and a fusion protein having the Fc region of the antibody or the like, a method for producing an antibody composition using the cell, the antibody composition and use thereof.
1. A non-human knock-out mammal lacking alpha-1,6-fucosylation, comprising an exogenous gene encoding an antibody molecule, wherein said mammal produces an antibody composition comprising the antibody molecule,
said antibody molecule comprising a Fc region comprising complex N-glycoside-linked sugar chains bound to the Fc region, and
said sugar chains comprising a reducing end which contains an N-acetylglucosamine, wherein the sugar chains do not contain fucose bound to the 6-position of the N-acetylglucosamine in the reducing end of the sugar chains,
wherein said lack of alpha-1,6-fucosylation results from inactivation of an endogenous gene encoding an enzyme selected from the group consisting of GDP-mannose 4,6-dehydratase (GMD), GDP-keto-6-deoxymannose 3,5-epimerase, 4-reductase (Fx), GDP-beta-L-fucose pyrophosphorylase (GFPP) and alpha-1,6-fucosyltransferase (FUT8), such that said non-human knock-out mammal lacking alpha-1,6-fucosylation produces said antibody molecule.
2. The mammal of claim 1 wherein said sugar chains comprise:
3. The mammal of claims 1 or 2 , wherein said mammal is selected from the group consisting of a cow, a sheep, a goat, a pig, a horse, a mouse, a rat, a monkey and a rabbit.
4. The mammal of claims 1 or 2 , wherein said antibody molecule is an IgG antibody molecule.
5. The mammal of claims 1 or 2 , wherein said antibody molecule is an antibody fragment.
6. An isolated knock-out mammalian cell lacking alpha-1,6-fucosylation, comprising an exogenous gene encoding an antibody molecule, wherein said mammalian cell produces an antibody composition comprising the antibody molecule,
said antibody molecule comprising a Fc region comprising complex N-glycoside-linked sugar chains bound to the Fc region,
said sugar chains comprising a reducing end which contains an N-acetylglucosamine, wherein the sugar chains do not contain fucose bound to the 6-position of the N-acetylglucosamine in the reducing end of the sugar chains,
said isolated knock-out mammalian cell lacking alpha-1,6-fucosylation is a cell in which a gene encoding an enzyme selected from the group consisting of GDP-mannose 4,6-dehydratase (GMD), GDP-keto-6-deoxymannose 3,5-epimerase, 4-reductase (Fx), GDP-beta-L-fucose pyrophosphorylase (GFPP) and alpha-1,6-fucosyltransferase (FUT8) is inactivated or substituted with a sequence, such that said isolated knock-out mammalian cell lacking alpha-1,6-fucosylation produces said antibody molecule.
7. The isolated mammalian cell of claim 6 wherein said sugar chains comprise:
8. The mammalian cell of claims 6 or 7 , wherein said mammalian cell is a mammalian cell selected from the group consisting of a cow, a sheep, a goat, a pig, a horse, a mouse, a rat, a monkey and a rabbit cell.
9. A method of producing an antibody molecule comprising an Fc region comprising N-glycoside-linked sugar chains which do not contain fucose at the 6-position of the N-acetylglucosamine of the reducing terminal of said sugar chains bound to the Fc region, said method comprising culturing a mammalian cell comprising an exogenous gene encoding an antibody molecule, to produce said antibody molecule;
wherein said mammalian cell is not capable of adding said fucose to the 6-position of the N-acetylglucosamine of the reducing terminal of said sugar chains bound to the Fc region of the antibody molecule,
wherein said mammalian cell is a cell in which a gene encoding an enzyme selected from the group consisting of GDP-mannose 4,6-dehydratase (GMD), GDP-keto-6-deoxymannose 3,5-epimerase, 4-reductase (Fx), GDP-beta-L-fucose pyrophosphorylase (GFPP) and alpha-1,6-fucosyltransferase (FUT8) is inactivated or substituted with a sequence, such that said mammalian cell lacks alpha-1,6-fucosylation.
10. The method of claim 9 wherein said sugar chain comprises:
11. The method of claim 9 wherein said mammalian cell is an alpha-1,6-fucosyltransferase knock-out cell.
12. A method of producing an antibody molecule comprising the Fc region comprising N-glycoside-linked sugar chains which do not contain fucose at the 6-position of the N-acetylglucosamine of the reducing terminal of said sugar chains bound to the Fc region, said method comprising maintaining a non-human mammal comprising an exogenous gene encoding an antibody molecule, to produce said antibody molecule;
wherein said mammal is not capable of adding said fucose to the 6-position of the N-acetylglucosamine of the reducing terminal of said sugar chain bound to the Fc region of the antibody molecule,
wherein said mammal is a mammal in which a gene encoding an enzyme selected from the group consisting of GDP-mannose 4,6-dehydratase (GMD), GDP-keto-6-deoxymannose 3,5-epimerase, 4-reductase (Fx), GDP-beta-L-fucose pyrophosphorylase (GFPP) and alpha-1,6-fucosyltransferase (FUT8) is inactivated or substituted with a sequence, such that said mammal lacks alpha-1,6-fucosylation.
13. The method of claim 12 wherein said sugar chain comprises:
14. The method of claim 12 wherein said mammal is an alpha-1,6-fucosyltransferase knock-out mammal.