Patent ID: 6642011
Filing Date: 2003-11-04
Classification: A61K,A61P,C12N

Abstract:
A method for producing a mutant protease having reduced allergenicity comprising the steps of:a) obtaining a naturally-occurring protease having subtilisin activity and preparing fragments of said naturally-occurring protease having subtilisin activity; b) contacting said fragments of said naturally-occurring protease with a first solution comprising naÃ¯ve human CD4+ or CD8+ T-cells and dendritic cells, wherein said dendritic cells have been differentiated; c) identifying an epitope region of said naturally-occurring protease, wherein said identifying comprises measuring the ability of said fragments of said naturally-occurring protease epitope region to stimulate proliferation of said naÃ¯ve human CD4+ or CD8+ T-cells; d) replacing said epitope region identified in step c) with an analogous epitope region in the amino acid sequence set forth in SEQ ID NO:6, to produce said mutant protease; e) preparing fragments of said mutant protease; f) contacting said fragments of said mutant protease with a second solution comprising naÃ¯ve human CD4+ or CD8+ T-cells and dendritic cells, wherein said dendritic cells have been differentiated; and g) measuring the ability of said fragments of said mutant protease to stimulate proliferation of said naÃ¯ve human CD4+ or CD8+ T-cells.