title: A novel approach for the detection of potentially hazardous pepsin stable hazelnut proteins as contaminants in chocolate-based food.
abstract: Contamination of food products with pepsin resistant allergens is generally believed to be a serious threat to patients with severe food allergy. A sandwich type enzyme-linked immunosorbent assay (ELISA) was developed to measure pepsin resistant hazelnut protein in food products. Capturing and detecting rabbit antibodies were raised against pepsin-digested hazelnut and untreated hazelnut protein, respectively. The assay showed a detection limit of 0.7 ng/mL hazelnut protein or <1 microg hazelnut in 1 g food matrix and a maximum of 0.034% cross-reactivity (peanut). Chocolate samples spiked with 0.5-100 microg hazelnut/g chocolate showed a mean recovery of 97.3%. In 9/12 food products labeled "may contain nuts", hazelnut was detected between 1.2 and 417 microg hazelnut/g food. It can be concluded that the application of antibodies directed to pepsin-digested food extracts in ELISA can facilitate specific detection of stable proteins that have the highest potential of inducing severe food anaphylaxis.
authors.affiliation: Sanquin Research, Department of Immunopathology and Laboratory for Experimental and Clinical Immunology, Academic Medical Centre, University of Amsterdam, 1066 CX Amsterdam, The Netherlands.
authors.forename: Jaap H
authors.initials: JH
authors.lastname: Akkerdaas
authors.forename: Marjolein
authors.initials: M
authors.lastname: Wensing
authors.forename: André C
authors.initials: AC
authors.lastname: Knulst
authors.forename: Oliver
authors.initials: O
authors.lastname: Stephan
authors.forename: Susan L
authors.initials: SL
authors.lastname: Hefle
authors.forename: Rob C
authors.initials: RC
authors.lastname: Aalberse
authors.forename: Ronald
authors.initials: R
authors.lastname: van Ree
mesh_terms: D002099:Cacao / Q000737:chemistry*; D031211:Corylus / Q000737:chemistry*; D004355:Drug Stability; D004797:Enzyme-Linked Immunosorbent Assay; D005506:Food Contamination / Q000032:analysis*; D010434:Pepsin A / Q000378:metabolism*; D010940:Plant Proteins / Q000032:analysis* / Q000378:metabolism
chemical_list: D010940:Plant Proteins; D010434:Pepsin A