Investigation of the stability of apple allergen extracts

To determine optimal conditions for allergen preservation, we investigated the influence of different stabilizing additives and of storage temperature on the allergen activity of apple protein preparations, obtained by extraction in phosphate buffer or by precipitation in diacetone alcohol and resolubilization in phosphate buffer in the presence or absence of enzyme inhibitors. For this purpose, the extracts were stored for 6 months either in frozen state at -20 degrees C or in lyophilized state at -20 degrees C, 4 degrees C, or room temperature and were characterized by SDS-PAGE, immunoblot, ELISA inhibition, and prick test. The highest stability revealed the extracts that were prepared by precipitation in the organic solvent in the presence of enzyme inhibitors, lyophilized, and stored at -20 degrees C. For storage of extract solutions at 4 degrees C, PBS/glycerol and cysteine/sodium citrate/glycerol were found to be the most effective stabilizing additives.