Production of high mannose proteins in plant culture and therapeutic uses thereof
A device, system and method for producing glycosylated proteins in plant culture, particularly proteins having a high mannose glycosylation, while targeting such proteins with an ER signal and/or by-passing the Golgi. The invention further relates to vectors and methods for expression and production of enzymatically active high mannose lysosomal enzymes using transgenic plant root, particularly carrot cells. More particularly, the invention relates to host cells, particularly transgenic suspended carrot cells, vectors and methods for high yield expression and production of biologically active high mannose Glucocerebrosidase (GCD). The invention further provides for compositions and methods for the treatment of lysosomal storage diseases.
1. A method for treating a subject having Gaucher's disease using exogenous human glucocerebrosidase protein, the method comprising administering a therapeutically effective amount of a glucocerebrosidase protein to said subject, wherein said human glucocerebrosidase protein is as set forth in SEQ ID NO: 15, thereby treating said Gaucher's disease in said subject.
2. The method of claim 1 , wherein said human glucocerebrosidase protein is glycosylated with at least one xylose and at least one exposed mannose residue.
3. The method of claim 1 , wherein said human glucocerebrosidase protein is glycosylated with at least one core α-(1,2) xylose and at least one core α-(1,3) fucose.
4. The method of claim 3 , wherein said human glucocerebrosidase protein is glycosylated with at least one exposed mannose residue.
5. The method of claim 1 , wherein said human glucocerebrosidase protein is glycosylated with at least one exposed mannose residue and at least one fucose residue having an alpha (1-3) glycosidic bond.
6. The method of claim 2 , wherein said xylose residue is a core α-(1,2) xylose residue.
7. The method of claim 2 , wherein said glucocerebrosidase protein binds to a mannose receptor on a macrophage.
8. The method of claim 4 , wherein said glucocerebrosidase protein binds to a mannose receptor on a macrophage.
9. The method of claim 7 , wherein said glucocerebrosidase is taken up into macrophages.
10. The method of claim 9 , wherein said human glucocerebrosidase protein having an increased uptake by said macrophages, in comparison with the corresponding macrophage uptake of a recombinant glucocerebrosidase expressed in mammalian cells.
11. The method of claim 6 , wherein the main glycan structure of said therapeutically effective amount of said human glucocerebrosidase protein is glycosylation with one exposed mannose residue.
12. A method of increasing hydrolysis of glycosphingolipid glucocerebroside in macrophages of a subject in need thereof, the method comprising administering a therapeutically effective amount of a human glucocerebrosidase protein to said subject, wherein said human glucocerebrosidase protein is as set forth in SEQ ID NO: 15, thereby increasing hydrolysis of glycosphingolipid glucocerebroside in macrophages of said subject.
13. The method of claim 12 , wherein said human glucocerebrosidase protein is glycosylated with at least one xylose and at least one exposed mannose residue.
14. The method of claim 12 , wherein said human glucocerebrosidase protein is glycosylated with at least one exposed mannose residue and at least one fucose residue having an alpha (1-3) glycosidic bond.
15. The method of claim 12 , wherein the main glycan structure of said therapeutically effective amount of said human glucocerebrosidase protein is glycosylation with one exposed mannose residue.