Treatment of alpha-galactosidase a deficiency

Abstract

The invention provides highly purified α-Gal A, and various methods for purifying it; α-Gal A preparations with altered charge and methods for making those preparations; α-Gal A preparations that have an extended circulating half-life in a mammalian host, and methods for making same; and methods and dosages for administering an α-Gal A preparation to a subject.

1 - 45 . (canceled) 
     
     
         46 . A human α-galactosidase A (α-Gal A) glycoprotein preparation, wherein the preparation comprises an average of about 1 to 2 mannose-6-phosphate residues per α-Gal A monomer and greater than 50% of the total glycans of the preparation are complex-type glycans. 
     
     
         47 . The preparation of  claim 46 , wherein the preparation is purified to at least 98% homogeneity, as measured by SDS-PAGE or reverse phase HPLC. 
     
     
         48 . The preparation of  claim 46 , wherein the preparation has a specific activity of at least 2.0×10 6  units/mg protein. 
     
     
         49 . The preparation of  claim 46 , wherein the preparation comprises an average of two complex glycans per α-Gal A monomer. 
     
     
         50 . The preparation of  claim 46 , wherein more than 50% of the total glycans of the preparation are sialyated. 
     
     
         51 . The preparation of  claim 46 , wherein the complex glycans have about 2 to 4 sialic acid residues. 
     
     
         52 . The preparation of  claim 46 , wherein about 15% of the total glycans of the preparation are neutral glycans. 
     
     
         53 . The preparation of  claim 46 , wherein about 16% of the total glycans of the preparation are phosphorylated glycans. 
     
     
         54 . The preparation of  claim 46 , wherein the preparation is isolated from human cells transfected with an α-Gal A/mammalian expression vector construct. 
     
     
         55 . A pharmaceutical composition comprising a human α-galactosidase A (α-Gal A) glycoprotein preparation, wherein the preparation comprises an average of about 1 to 2 mannose-6-phosphate residues per α-Gal A monomer and greater than 50% of the total glycans of the preparation are complex-type glycans. 
     
     
         56 . The composition of  claim 55 , wherein the composition comprises one unit dose between 0.1-0.3 mg of the preparation per kilogram body weight. 
     
     
         57 . The composition of  claim 55 , wherein the preparation is purified to at least 98% homogeneity, as measured by SDS-PAGE or reverse phase HPLC. 
     
     
         58 . The composition of  claim 55 , wherein the preparation has a specific activity of at least 2.0×10 6  units/mg protein. 
     
     
         59 . The composition of  claim 55 , wherein the preparation comprises an average of two complex glycans per α-Gal A monomer. 
     
     
         60 . The composition of  claim 55 , wherein more than 50% of the total glycans of the preparation are sialyated. 
     
     
         61 . The composition of  claim 55 , wherein the complex glycans have about 2 to 4 sialic acid residues. 
     
     
         62 . The composition of  claim 55 , wherein about 15% of the total glycans of the preparation are neutral glycans. 
     
     
         63 . The composition of  claim 55 , wherein about 16% of the total glycans of the preparation are phosphorylated glycans. 
     
     
         64 . The composition of  claim 55 , wherein the preparation is isolated from human cells transfected with an α-Gal A/mammalian expression vector construct. 
     
     
         65 . A method of treating a subject, comprising administering to a subject in need thereof a human α-galactosidase A (α-Gal A) glycoprotein preparation, wherein the preparation comprises an average of about 1 to 2 mannose-6-phosphate residues and greater than 50% of the total glycans of the preparation are complex-type glycans. 
     
     
         66 . The method of  claim 65 , wherein the preparation is administered at a dose between 0.1-0.3 mg per kilogram body weight. 
     
     
         67 . The method of  claim 65 , wherein the preparation is administered weekly or biweekly. 
     
     
         68 . The method of  claim 65 , wherein the preparation is purified to at least 98% homogeneity, as measured by SDS-PAGE or reverse phase HPLC. 
     
     
         69 . The method of  claim 65 , wherein the preparation has a specific activity of at least 2.0×10 6  units/mg protein. 
     
     
         70 . The method of  claim 65 , wherein the preparation comprises an average of two complex glycans per α-Gal A monomer. 
     
     
         71 . The method of  claim 65 , wherein more than 50% of the total glycans of the preparation are sialyated. 
     
     
         72 . The method of  claim 65 , wherein the complex glycans have about 2 to 4 sialic acid residues. 
     
     
         73 . The method of  claim 65 , wherein about 15% of the total glycans of the preparation are neutral glycans. 
     
     
         74 . The method of  claim 65 , wherein about 16% of the total glycans of the preparation are phosphorylated glycans. 
     
     
         75 . The method of  claim 65 , wherein the preparation is isolated from human cells transfected with an α-Gal A/mammalian expression vector construct.