Method of producing recombinant antithrombin III composition
View Patent ↗The present invention provides a process for producing an antithrombin III composition comprising an antithrombin III molecule having complex type N-glycoside-linked sugar chains, wherein the complex type N-glycoside-linked sugar chains have a structure in which fucose is not bound to N-acetylglucosamine in the reducing end in the sugar chains.
1. An isolated, recombinant antithrombin III in which an amino acid sequence of the recombinant antithrombin III consists of the amino acid sequence at positions from 33 to 464 of SEQ ID NO:4;
wherein the recombinant antithrombin III comprises four complex type N-glycoside-linked carbohydrate chains each at asparagines residues at positions 128, 167, 187 and 224 in the amino acid sequence of SEQ ID NO:4, wherein each of said carbohydrate chains lacks fucose bound to N-acetylglucosamine in the reducing end of said carbohydrate chains,
wherein the recombinant antithrombin III is expressed in a CHO cell into which a DNA encoding the amino acid sequence at positions from 33 to 464 of SEQ ID NO:4 is introduced, wherein the genome of the CHO cell is modified so as to have deleted activity of an enzyme relating to synthesis of an intracellular carbohydrate nucleotide, GDP-fucose, or an enzyme relating to the modification of a carbohydrate chain in which 1-position of fucose is bound to 6-position of N-acetylglucosamine in the reducing end through α-bond in a complex type N-glycoside-linked carbohydrate chain.
2. A medicament which comprises the antithrombin III according to claim 1 as an active ingredient and a pharmaceutically acceptable carrier.
3. The medicament according to claim 2 which has an activity of inhibiting blood coagulation.