Method for secretory production of glycoprotein having human-type sugar chain using plant cell

Abstract

A method for the secretory production of a glycoprotein having a human-type sugar chain, comprising a step of introducing a gene of an enzyme capable of performing a transfer reaction of a galactose residue to a non-reducing terminal acetylglucosamine residue, and a gene of heterologous glycoprotein, to obtain a transformed plant cell, a step of culturing the plant cell, and a step of recovering the culture medium of the plant cell.

1 . A method for the secretory production of a glycoprotein having a human-type sugar chain, comprising: 
 a step of introducing a gene of an enzyme capable of performing a transfer reaction of a galactose residue to a non-reducing terminal acetylglucosamine residue and a gene of a heterologous glycoprotein, to obtain a transformed plant cell, and    a step of culturing said plant cell.    
     
     
         2 . The method according to  claim 1 , wherein said glycoprotein having a human-type sugar chain comprises a core sugar chain and an outer sugar chain, said core sugar chain substantially comprises a plurality of mannoses and acetylglucosamines, and said outer sugar chain has a terminal sugar chain moiety containing a non-reducing terminal galactose.  
     
     
         3 . The method according to  claim 2 , wherein said outer sugar chain has a linear structure.  
     
     
         4 . The method according to  claim 2 , wherein said outer sugar chain has a branched structure.  
     
     
         5 . The method according to  claim 4 , wherein said branched structure is a mono-, bi-, tri- or tetra-structure.  
     
     
         6 . The method according to  claim 1 , wherein said glycoprotein is free of fucose or xylose.  
     
     
         7 . The method according to  claim 1 , which further comprises a step of recovering the culture solution of said plant cells.  
     
     
         8 . The method according to  claim 1 , which further comprises a step of adding sugar or a sugar chain in vitro.  
     
     
         9 . A plant cell which comprises a sugar chain adding mechanism capable of performing a transfer reaction of a galactose residue to a non-reducing terminal acetylglucosamine residue and can secrete a protein with an added sugar chain added by said sugar chain-adding mechanism, wherein said sugar chain-adding mechanism adds a sugar chain comprising a core sugar chain and an outer sugar chain, said core sugar chain substantially comprises a plurality of mannoses and acetylglucosamines, and said outer sugar chain has a terminal sugar chain moiety containing a non-reducing terminal galactose.  
     
     
         10 . A glycoprotein having a human-type sugar chain obtained by the method according to  claim 1 .  
     
     
         11 . A method for the secretory production of a glycoprotein having human-type sugar chain, comprising: 
 a step of introducing a gene for an enzyme capable of performing a transfer reaction of a galactose residue to a non-reducing terminal acetylglucosamine residue and a gene for a heterologous glycoprotein, to obtain a transformed plant cell, and    a step of expressing said enzyme within an intracellular organelle.    
     
     
         12 . A plant cell transformed with a gene for an enzyme capable of performing a transfer reaction of a galactose residue to a non-reducing terminal acetylglucosamine residue, 
 wherein said enzyme is localized in the plant cell such that the plant cell is capable of synthesizing a glycoprotein having a human-type sugar chain structure.    
     
     
         13 . A plant cell according to  claim 12 , wherein said sugar chain structure includes an added galactose residue.  
     
     
         14 . A plant cell according to  claim 13 , wherein said sugar chain structure is free of β1,2-xylose or α1,3-fucose.  
     
     
         15 . A plant cell according to  claim 12 , wherein said sugar chain structure includes an galactose residue added to a N-linked type sugar chain of (N-acetylglucosamine) 1-2 (Mannose) 2-5 (N-acetylglucosamine) 2  selected from the group consisting of GlcNAc 1 Man 3 GlcNAc 2 , GlcNAc 1 Man 5 GlcNAc 2 , GlcNAc 2 Man 3 GlcNAc 2  and GlcNAc 1 Man 4 GlcNAc 2 .  
     
     
         16 . A plant cell according to  claim 12 , wherein said enzyme is localized within an intracellular organelle in the plant cell.  
     
     
         17 . A plant regenerated from the plant cell of  claim 12 .  
     
     
         18 . A seed produced from the plant of  claim 16 .  
     
     
         19 . The method according to  claim 5 , wherein said glycoprotein is free of fucose or xylose.  
     
     
         20 . The method according to  claim 5 , which further comprises a step of adding sugar or a sugar chain in vitro.  
     
     
         21 . A glycoprotein having a human-type sugar chain obtained by the method according to  claim 5.