Enhanced liquid formulation stability of erythropoietin alpha through purification processing

Abstract

A method for purifying rHuEPO through the use of a multi-step filtration process which provides for a rHuEPO product having superior stability and shelf-life.

What is claimed is: 
     
         1 . A recombinant human erythropoietin alpha product having a shelf life of at least four years manufactured by the following process wherein the process comprises the following steps:
 a. manufacturing a recombinant human erythropoietin alpha product and collecting the supernatant,   b. clarifying the supernatant and subjecting the supernatant to a multi-step purification process comprising the following procedures in order: ultradiafiltration, anion exchange chromatography, ammonium sulfate precipitation, hydrophobic interaction chromatography, acetonitrile addition, reversed phase chromatography, a second anion exchange chromatography step, gelfiltration, and nano filtration for virus removal; and   c. producing a recombinant human erythropoietin alpha product that is substantially free of non-O-glycosylated recombinant human erythropoietin alpha isoforms.   
     
     
         2 . The product made by the process of  claim 1 , wherein said purified human erythropoietin alpha product is filled into a Type 1 glass container. 
     
     
         3 . The product made by the process of  claim 1  wherein step b wherein the urea in a wash buffer in the first anion exchange chromatography step is at a concentration of less than 3M. 
     
     
         4 . The product made by the process of  claim 1  wherein step b wherein the urea in a wash buffer in the first anion exchange chromatography step is at a concentration of less than 0.1M.