Methods for purification of arylsulfatase a

Abstract

Methods of producing arylsulfatase A are described herein. The methods can include one or more steps of chromatography. Exemplary chromatographic steps include ion exchange chromatography, mixed mode chromatography, and hydrophobic interaction chromatography. Compositions containing and methods using arylsulfatase A produced by the methods described herein are also disclosed.

1 .- 36 . (canceled) 
     
     
         37 . A method comprising
 providing a sample of recombinant arylsulfatase A (ASA);   purifying the recombinant ASA protein from one or more contaminants by subjecting the sample to ion exchange chromatography, mixed-mode chromatography, and hydrophobic interaction chromatography.   
     
     
         38 . The method of  claim 37 , wherein the purified recombinant ASA protein has a specific activity of at least about 75 U/mg. 
     
     
         39 . The method of  claim 37 , wherein the purified recombinant ASA protein contains less than 1.5% whole cell proteins. 
     
     
         40 . The method of  claim 37 , wherein the sample of recombinant ASA protein is obtained from crude cell extract or cell culture supernatant. 
     
     
         41 . The method of  claim 37 , wherein the method comprises subjecting the sample of recombinant ASA to a first and a second ion exchange chromatography, mixed-mode chromatography, and hydrophobic interaction chromatography. 
     
     
         42 . The method of  claim 41 , wherein the first ion exchange chromatography is anion-exchange chromatography. 
     
     
         43 . The method of  claim 42 , wherein the anion exchange chromatography is quaternary amine anion exchange chromatography. 
     
     
         44 . The method of  claim 41 , wherein the second ion exchange chromatography is cation exchange chromatography. 
     
     
         45 . The method of  claim 37 , wherein the mixed-mode chromatography is hydroxyapatite (HA) chromatography. 
     
     
         46 . The method of  claim 37 , wherein the hydrophobic interaction chromatography is phenyl chromatography. 
     
     
         47 . The method of  claim 41 , wherein the method comprises subjecting the sample of recombinant ASA to a first ion exchange chromatography, subjecting the first ion exchange chromatography sample to mixed-mode chromatography, subjecting the mixed-mode chromatography sample to hydrophobic interaction chromatography, and subjecting the hydrophobic interaction chromatography sample to a second ion exchange chromatography, thereby obtaining purified recombinant ASA protein. 
     
     
         48 . The method of  claim 47 , wherein the first ion exchange chromatography is anion exchange chromatography. 
     
     
         49 . The method of  claim 47 , wherein the second ion exchange chromatography is cation exchange chromatography. 
     
     
         50 . The method of  claim 47 , wherein the method further comprises subjecting the sample of recombinant ASA to viral inactivation before loading the sample onto the first ion exchange chromatography column. 
     
     
         51 . A pharmaceutical composition comprising a recombinant ASA protein purified according to a method of  claim 37 . 
     
     
         52 . A method of treating metachromatic leukodystrophy (MLD) in a subject, comprising administering to the subject a pharmaceutical composition of  claim 51 . 
     
     
         53 . A formulation comprising a recombinant ASA protein purified according to a method of  claim 37 , wherein the formulation contains less than 1.5% whole cell proteins. 
     
     
         54 . The formulation of  claim 53 , wherein the formulation is formulated for intravenous delivery. 
     
     
         55 . The formulation of  claim 53 , wherein the formulation is formulated for intrathecal delivery. 
     
     
         56 . A method of treating metachromatic leukodystrophy (MLD) in a subject, comprising administering to the subject a formulation of  claim 53 .