FIG. 2.

IgE binding of synthetic peptides of Asp f 2 with sera from ABPA patients. Linear decapeptides were synthesized on derivatized cellulose membranes with an offset of 1 aa. On each spot on the membrane marked for peptide synthesis, a free amino functional group was available for coupling with the carboxyl group of the N-terminal amino acid of the peptide. Ten cycles of amino acid addition were carried out for synthesis of decapeptides. After completion of the peptide synthesis, the protecting groups on the side chains of the amino acids were removed. The IgE binding epitopes were identified at various regions of Asp f 2 based on the antibody peptide reaction demonstrated by enzyme immunostaining (for details, see Materials and Methods).