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J Immunol Methods. 1989 Jul 26;121(2):275-80.

A method for the detection of IgE binding sequences of allergens based on a modification of epitope mapping.

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  • 1School of Chemistry, Macquarie University, N.S.W., Australia.


An ELISA method for the rapid determination of IgE binding sites (allergenic determinants) of proteins is reported. The method utilizes the epitope mapping kit (Geysen et al., 1984) to synthesize hexapeptides of an allergen of interest, followed by a biotin-avidin system to detect peptide-bound IgE. The technique allows rapid localisation of determinants from allergens of known sequence without the need to purify large amounts of allergen nor to generate peptides by cleavage of it. Using the results of the epitope mapping experiments a putative allergenic peptide containing 18 amino acid residues from the sequence of a wheat allergen was identified and synthesised on polyamide resin. Testing of this peptide by radioallergosorbent test (RAST) inhibition showed that it bound specific IgE in the sera of patients allergic to wheat.

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