Human lysosomal proteins from plant cell culture

Abstract

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.

What is claimed is: 
     
         1 . A pharmaceutical composition for treatment or prevention of a disease associated with a deficiency in a lysosomal enzyme function comprising lyophilized plant cells comprising a recombinant lysosomal enzyme protein and a pharmaceutically acceptable carrier. 
     
     
         2 . The pharmaceutical composition of  claim 1 , wherein said recombinant lysosomal enzyme is selected from the group consisting of alpha galactosidase, ceramidase, beta galactosidase, beta hexosaminidase, sphingomyelinase, alpha-N-acetylgalactosaminidase, iduronate-2-sulfatase, beta-glucoronidase, iduronidase and mannose-6-phosphate transporter. 
     
     
         3 . The pharmaceutical composition of  claim 1 , wherein said recombinant human lysosomal enzyme is glycosylated. 
     
     
         4 . The pharmaceutical composition of  claim 1 , wherein said plant cell is a carrot cell. 
     
     
         5 . The pharmaceutical composition of  claim 4 , wherein said plant cell is a carrot cell from suspension culture. 
     
     
         6 . The pharmaceutical composition of  claim 1  formulated for oral administration. 
     
     
         7 . The pharmaceutical composition of  claim 1 , wherein said recombinant human lysosomal enzyme has lysosomal enzyme biological activity. 
     
     
         8 . A composition of matter comprising lyophilized plant cells comprising a recombinant human lysosomal enzyme protein. 
     
     
         9 . The composition of matter of  claim 8 , wherein said human lysosomal enzyme protein is selected from the group consisting of alpha galactosidase, ceramidase, beta galactosidase, beta hexosaminidase, sphingomyelinase, alpha-N-acetylgalactosaminidase, iduronate-2-sulfatase, beta-glucoronidase, iduronidase and mannose-6-phosphate transporter. 
     
     
         10 . The composition of matter of  claim 8 , wherein said plant cell is a carrot cell. 
     
     
         11 . The composition of matter of  claim 8 , wherein said plant cell is a carrot cell from suspension culture. 
     
     
         12 . The composition of matter of  claim 8  formulated for oral administration. 
     
     
         13 . The composition of matter of  claim 8 , wherein said recombinant human lysosomal enzyme protein has lysosomal enzyme catalytic activity. 
     
     
         14 . A method of treating or preventing a disease associated with a deficiency in a lysosomal enzyme function in a subject comprising orally administering lyophilized plant cells comprising a recombinant lysosomal enzyme protein. 
     
     
         15 . The method of  claim 14 , wherein said disease is selected from the group consisting of Fabry disease, Farber disease, G ml  gangliosidosis, Tay-Sachs disease, Niemann-Pick disease, Schindler disease, Hunter syndrome, Sly syndrome, Hurler and Hurler/Scheie syndromes, and I-Cell/San Filipo syndrome. 
     
     
         16 . The method of  claim 14 , wherein said disease is Fabry disease and said recombinant lysosomal enzyme protein is alpha-galactosidase. 
     
     
         17 . The method of  claim 14 , wherein said disease is Farber disease and said recombinant lysosomal enzyme protein is ceramidase. 
     
     
         18 . The method of  claim 14 , wherein said disease is G ml  gangliosidosis and said recombinant lysosomal enzyme protein is beta-galactosidase. 
     
     
         19 . The method of  claim 14 , wherein said disease is Tay-Sachs disease and said recombinant lysosomal enzyme protein is beta-hexosaminidase. 
     
     
         20 . The method of  claim 14 , wherein said disease is Niemann-Pick disease and said recombinant lysosomal enzyme protein is sphingomyelinase. 
     
     
         21 . The method of  claim 14 , wherein said disease is Schindler disease and said recombinant lysosomal enzyme protein is alpha.-N-acetylgalactosaminidase. 
     
     
         22 . The method of  claim 14 , wherein said disease is Hunter syndrome and said recombinant lysosomal enzyme protein is iduronate-2-sulfatase. 
     
     
         23 . The method of  claim 14 , wherein said disease is Sly syndrome and said recombinant lysosomal enzyme protein is beta-glucuronidase. 
     
     
         24 . The method of  claim 14 , wherein said disease is Hurler or Hurler/Scheie syndrome and said recombinant lysosomal enzyme protein is iduronidase. 
     
     
         25 . The method of  claim 14 , wherein said disease is I-Cell/San Filipo syndrome and said recombinant lysosomal enzyme protein is mannose 6-phosphate transporter.