Human protease and use of such protease for pharmaceutical applications and for reducing the allergenicity of non-human proteins

Abstract

The present invention relates to a method of producing novel improved protein mutant which produce low allergenic response in humans compared to the parent of that mutant. Specifically, the present invention comprises neutralizing or reducing the allergenicity of a protein by introducing therein as replacement or modification of an epitope on such protein a sequence from human subtilisin.

We claim:  
     
         1 . A method for reducing the allergenicity of a non-human protein is provided wherein an epitope is identified and replaced with an analogous region within a human subtilisin.  
     
     
         2 . The method according to  claim 1 , wherein said non-human protein comprises a protease.  
     
     
         3 . The method according to  claim 2 , wherein said epitope replaced is a T-cell epitope.  
     
     
         4 . A method for producing the protein of the invention having reduced allergenicity comprising, preparing a mutant protein by modifying a DNA encoding a precursor protein so that the modified DNA encodes the mutant protein of the invention, wherein an epitope is replaced with an analogous region from human subtilisin.  
     
     
         5 . A mutant protein having reduced allergenicity produced according to the method of  claim 4 .  
     
     
         6 . DNA encoding the mutant protein of  claim 5 .  
     
     
         7 . An expression vector containing a DNA sequence according to  claim 6 .  
     
     
         8 . Host cells transformed with the vectors of  claim 7 .  
     
     
         9 . A method of producing a protein having reduced allergenicity wherein the host cells of  claim 8  are cultivated under conditions which result in expression of the mutant protease.  
     
     
         10 . A pharmaceutical composition comprising a human subtilisin.  
     
     
         11 . A DNA encoding a human subtilisin.  
     
     
         12 . A vector comprising the DNA of  claim 11 .  
     
     
         13 . A host cell comprising the vector of  claim 12 .  
     
     
         14 . A human subtilisin produced by the host cell of  claim 12.