Mutant proteins having lower allergenic response in humans and methods for constructing, identifying and producing such proteins

Abstract

The present invention relates to a novel improved protein mutant which produces low allergenic response in humans compared to the parent of that mutant. Specifically, the present invention comprises neutralizing or reducing the ability of T-cells to recognize epitopes and thus prevent sensitization of an individual to the protein.

1 - 11 . (canceled)  
     
     
         12 . A method for determining T-cell epitopes in a protein of interest comprising the steps of: 
 (a) obtaining from a single blood source a solution of dendritic cells and a solution of naive CD4+ T-cells;    (b) promoting differentiation in said solution of dendritic cells, by exposing said dendritic cells to at least one cytokine;    (c) combining said solution of differentiated dendritic cells and said naive CD4+ T-cells with a peptide of interest;    (d) measuring the proliferation of said T-cells in said step (c).    
     
     
         13 . A method of reducing the immunogenicity of a protein comprising the steps of: 
 (a) identifying at least one T-cell epitope in said protein, using the method of  claim 12;     (b) modifying said protein to neutralize said at least one T-cell epitope.    
     
     
         14 . The method according to  claim 13 , wherein said epitope is modified by: P 1  (a) substituting the amino acid sequence of the epitope with an analogous sequence from a homolog of the protein of interest wherein said analogous sequence produces a lower response from said T-cells than that of the protein of interest; or 
 (b) substituting the amino acid sequence of the epitope with a sequence which substantially mimics the major tertiary structure attributes of the epitope, but which produces a lowere response from T-cells than that of the protein of interest.    
     
     
         15 . A protein having reduced immunogenicity produced using the method of  claim 14 .  
     
     
         16 . A protein having reduced immunogenicity, wherein said protein comprises a modification comprising the substitution or deletion of amino acid residues which are identified as being within a T-cell epitope identified using the method of  claim 13.