Sta c 3 epitopes and their application as biomarkers to detect specific IgE

Sta c 3 was determined to be one of the major allergens from Stachybotrys chartarum sensu lato. This is among the most common fungi on wet building materials. Various cognizant authorities have found that mold and dampness are associated with exacerbation of asthma and increased upper respiratory disease. The primary amino-acid sequence of this allergen was reported, however the IgE-binding epitopes were unknown. Using SPOT-synthesis techniques (SPOTs), one main linear epitope located between V91 and G105 was discovered by Western blot with atopic human sera. This was confirmed by inhibition ELISA assays with both synthesized epitopes and natural Sta c 3. Alanine scanning also revealed R100 and K101 are the critical amino acids for IgE binding. Using the binding-enhanced and alanine substituted epitopes as potential biomarkers, a reverse ELISA method was developed to detect the Sta c 3 sIgE in atopic human sera. This method employed streptavidin-biotinylated mutated epitopes as the capture matrix, probing the possibility of using small peptides for allergen/species specific IgE assays.