Method for Preventing Formation of Trisulfide Derivatives of Polypeptides

Abstract

A method for reducing or substantially preventing formation of a trisulfide derivative of a polypeptide in a liquid medium containing the polypeptide ijn question comprises stripping the liquid medium with a gas, suitably a chemically unreactive gas such as nitrogen or argon.

1 . A method for reducing or substantially preventing formation of a trisulfide derivative of a polypeptide in a liquid medium containing said polypeptide, the method comprising stripping said liquid medium with a gas. 
     
     
         2 . A method according to  claim 1 , wherein said polypeptide contains one or more disulfide bonds. 
     
     
         3 . A method according to  claim 2 , wherein said polypeptide is a recombinantly produced polypeptide. 
     
     
         4 . A method according to  claim 3 , wherein said polypeptide is selected from: recombinantly produced human growth hormone (rhGH); truncated forms thereof; analogues thereof; and derivatives thereof. 
     
     
         5 . A method according to  claim 4 , wherein said stripping is performed during production or post-production processing of said polypeptide. 
     
     
         6 . A method according to  claim 3 , wherein said stripping is performed during post-production processing of said polypeptide. 
     
     
         7 . A method according to  claim 1 , wherein said gas is a chemically substantially inert gas. 
     
     
         8 . A method according to  claim 1 , wherein said gas is selected from the group consisting of nitrogen (N 2 ), helium (He), and argon (Ar). 
     
     
         9 . A method according to  claim 1 , wherein said polypeptide is subjected to one or more purification steps after the stripping procedure. 
     
     
         10 . A method according to  claim 9 , comprising at least one filtration step. 
     
     
         11 . A method according to  claim 5 , wherein a sulfite or a mercapto compound is added to the liquid medium after production of said polypeptide. 
     
     
         12 . A polypeptide substantially free of TS derivatives produced by a method according to  claim 1 . 
     
     
         13 . A recombinant human growth hormone (rhGH) product substantially free of TS-hGH derivatives, produced by a method according to  claim 4 . 
     
     
         14 . A pharmaceutical composition comprising rhGH according to  claim 13  together with a pharmaceutically acceptable carrier or diluent. 
     
     
         15 . A method for treating a condition responsive to administration of human growth hormone, comprising administering a therapeutically effective amount of substantially TS-hGH-free rhGH according to  claim 13  to a subject having such a condition. 
     
     
         16 . (canceled) 
     
     
         17 . A method according to  claim 4 , wherein said gas is a chemically substantially inert gas. 
     
     
         18 . A method according to  claim 4 , herein said gas is selected from the group consisting of nitrogen (N 2 ), helium (He), and argon (Ar). 
     
     
         19 . A method according to  claim 4 , wherein said polypeptide is subjected to one or more purification steps after the stripping procedure. 
     
     
         20 . A method according to  claim 4 , comprising at least one filtration step. 
     
     
         21 . A method according to  claim 6 , wherein a sulfite or a mercapto compound is added to the liquid medium after production of said polypeptide.