Preparation of Deallergenized Proteins and Permuteins

A method for decreasing allergen eliciting properties of a native protein comprising the steps of:

• a) identifying a patient exhibiting an allergic sensitivity to said native protein and obtaining serum from said patient;
• b) exposing synthetic overlapping peptides representative of said native protein to said patient serum to identify peptides exhibiting epitopes which bind IgE present within said patient serum;
• c) producing variant peptides exhibiting alanine scanning or rational scanning amino acid substitutions based on peptides from step (b), wherein said variant peptides exhibit decreased IgE binding compared to peptides from step (b), said amino acid substitutions comprising result effective substitutions;
• d) modifying the amino acid sequence of said native protein to contain one or more of said result effective substitutions, thereby obtaining a modified protein;
• and
• e) isolating and purifying the modified protein comprising said one or more result effective amino acid substitutions;
• wherein the modified protein exhibits decreased allergen eliciting properties of said native protein as determined by reduced binding of IgE when exposed to said patient serum when compared with said native protein.

A method for producing a modified acyl lipid hydrolase protein comprising:

• a) identifying a patient exhibiting an allergic sensitivity to a native acyl lipid hydrolase protein and obtaining serum from said patient;
• b) exposing synthetic overlapping peptides representative of said native acyl lipid hydrolase protein to said patient serum to identify peptides exhibiting epitopes which bind immunoglobulins present within said patient serum, said immunoglobulins exhibiting a binding specificity for said native acyl lipid hydrolase protein;
• c) producing variant peptides exhibiting alanine scanning or rational scanning amino acid substitutions based on peptides from step (b), wherein said variant peptides exhibit decreased immunoglobulin binding compared to peptides from step (b), said amino acid substitutions comprising result effective substitutions;
• d) modifying the amino acid sequence of said native acyl lipid hydrolase protein to contain one or more of said result effective substitutions, thereby obtaining a modified acyl lipid hydrolase protein;
• and
• e) isolating and purifying the modified acyl lipid hydrolase protein comprising said one or more result effective amino acid substitutions;
• wherein the modified acyl lipid hydrolase protein exhibits reduced binding of immunoglobulins when exposed to said patient serum when compared with said native acyl lipid hydrolase protein.