Method for manufacturing glycoproteins having human-type glycosylation

Abstract

The present invention provides a method for manufacturing a glycoprotein having a human-type sugar chain comprising a step in which transformed plant cell is obtained by introducing to a plant cell the gene of glycosyltransferase and the gene of an exogenous glycoprotein, and a step in which the obtained transformed plant cell is cultivated.

1 . A plant cell comprising a gene encoding a glycosyltransferase and a gene encoding an exogenous glycoprotein, in which the exogenous glycoprotein is expressed and the expressed glycoprotein comprises a human-type sugar chain. 
     
     
         2 . The plant cell according to  claim 1 , wherein the glycosyltransferase is galactosyltransferase that transfers a galactose residue to a non-reducing terminal acetylglucosamine residue. 
     
     
         3 . The plant cell according to  claim 2 , wherein the glycoprotein with a human-type sugar chain comprises a core sugar chain and an outer sugar chain, wherein the core sugar chain comprises a plurality of mannose and acetylglucosamine, and wherein the outer sugar chain contains a terminal sugar chain portion with a non-reducing terminal galactose. 
     
     
         4 . The plant cell according to  claim 3 , wherein the outer sugar chain has a straight chain configuration. 
     
     
         5 . The plant cell according to  claim 3 , wherein the outer sugar chain has a branched chain configuration. 
     
     
         6 . The plant cell according to  claim 5 , wherein the branched sugar chain portion has a mono-, bi-, tri, or tetra-configuration. 
     
     
         7 . The plant cell according to  claim 3 , wherein the glycoprotein produced contains no fucose or xylose linked to one or more of the core sugar chain, the outer sugar chain and the terminal sugar chain. 
     
     
         8 . The plant cell according to  claim 1 , wherein the gene encoding a glycosyl transferase encodes human galactosyltransferase, human galactosidase or human 3-galactosidase and the gene encodes an exogenous glycoprotein that is an enzyme, a hormone, a cytokine, an antibody, a vaccine, a receptor or a serum protein. 
     
     
         9 . The plant cell according to  claim 8 , wherein the exogenous glycoprotein encoded by the introduced gene is horseradish peroxidase, kinase, glucocerebrosidase, α-galactosidase, tissue-type plasminogen activator (TPA), or 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase. 
     
     
         10 . The plant according to  claim 8 , wherein the exogenous glycoprotein encoded by the introduced gene is enkephalin, interferon-alpha, granulocyte-macrophage colony stimulating factor (GM-CSF), granulocyte colony stimulating factor (C-CSF), chorion, stimulating hormone, interleukin-2, interferon-beta, interferon-gamma, erythropoietin, vascular endothelial growth factor, human choriogonadotropin (HCG), leuteinizing hormone (LH), thyroid stimulating hormone (TSH), prolactin, or ovary stimulating hormone. 
     
     
         11 . The plant cell according to  claim 8 , wherein the exogenous glycoprotein encoded by the introduced gene is immunoglobulin G (IgG) or single chain variable region antibody fragments (scFV). 
     
     
         12 . A plant cell comprising a glycoprotein comprising a human-type sugar chain, comprising no fucose or xylose linked to one or more of the core sugar chain, the outer sugar chain and the terminal sugar chain. 
     
     
         13 . A plant cell comprising an exogenous glycoprotein that comprises a galactose residue linked to an N-acetylglucosamine residue by a beta 1,4 linkage. 
     
     
         14 . A plant cell comprising an exogenous glycoprotein that comprises a galactose residue linked through the activity of a galactosyltransferase.