The present invention provides highly phosphorylated lysosomal hydrolases, methods of modifying lysosomal hydrolases with the lysosomal targeting pathway enzymes GlcNAc-phosphotransferase and/or phosphodiester α-GlcNAcase.

Methods for producing highly phosphorylated lysosomal hydrolases

The present invention involves the production of large quantities of human α-Gal A by cloning and expressing the α-Gal A coding sequence in eukaryotic host cell expression systems. The eukaryotic expression systems, and in particular the mammalian host cell expression system described herein provide for the appropriate cotranslational and posttranslational modifications required for proper processing, e.g., glycosylation, phosphorylation, etc. and sorting of the expression product so that an active enzyme is produced. In addition, the expression of fusion proteins which simplify purification is described. Using the methods described herein, the recombinant α-Gal A is secreted by the engineered host cells so that it is recovered from the culture medium in good yield. The α-Gal A produced in accordance with the invention may be used, but is not limited to, in the treatment in Fabry Disease; for the hydrolysis of α-galactosyl residues in glycoconjugates; and/or for the conversion of the blood group B antigen on erythrocytes to the blood group O antigen.

Cloning and expression of biologically active α-galactosidase A as a fusion protein

The present invention involves the production of large quantities of human alpha -Gal A by cloning and expressing the alpha -Gal A coding sequence in eukaryotic host cell expression systems. The eukaryotic expression systems, and in particular the mammalian host cell expression system described herein provide for the appropriate cotranslational and posttranslational modifications required for proper processing, e.g., glycosylation, phosphorylation, etc. and sorting of the expression product so that an glycosylation, phosphorylation, etc. and sorting of the expression product so that an active enzyme is produced. In addition, the expression of fusion proteins which simplify purification is described. Using the methods described herein, the recombinant alpha -Gal A is secreted by the engineered host cells so that it is recovered from the culture medium in good yield. The alpha -Gal A produced in accordance with the invention may be used in the treatment in Fabry Disease; for the hydrolysis of alpha -galactosyl residues in glycoconjugates; and/or for the conversion of the blood group B antigen on erythrocytes to the blood group O antigen.

Cloning and expression of biologically active human alpha -galactosidase A

The present invention involves the production of large quantities of human alpha-Gal A by cloning and expressing the alpha-Gal A coding sequence in eukaryotic host cell expression systems. The eukaryotic expression systems, and in particular the mammalian host cell expression system described herein provide for the appropriate cotranslational and posttranslational modifications required for proper processing, e.g., glycosylation, phosphorylation, etc. and sorting of the expression product so that an active enzyme is produced. In addition, the expression of fusion proteins which simplify purification is described. Using the methods described herein, the recombinant alpha-Gal A is secreted by the engineered host cells so that it is recovered from the culture medium in good yield. The alpha-Gal A produced in accordance with the invention may be used, but is not limited to, in the treatment in Fabry Disease; for the hydrolysis of alpha-galactosyl residues in glycoconjugates; and/or for the conversion of the blood group B antigen on erythrocytes to the blood group O antigen.

Cloning and expression of biologically active alpha-galactosidase a

The present invention provides methods of treating lysosomal storage disease with highly phosphorylated lysosomal hyrdrolases.

Methods of treating lysosomal storage diseases

A process for the production of α-galactosidase by culturing the mold Penicillium duponti in an aqueous medium containing at least one sugar with at least one αD-galactopyranosyl bond and collecting the mycelium thus obtained.

Arnaud Nicole

Papers

Alpha-galactosidase production