BackgroundRegulations in many countries worldwide prescribe that peanut must be listed on food labels as a cause of food allergies. Re-evaluated voluntary incidental trace allergen labelling (VITAL) values for peanut revealed the eliciting dose (ED01) value—at which 99% of all peanut-allergic individuals will not react—is 0.2 mg peanut protein.ObjectiveValidation of a sandwich ELISA based on monoclonal antibodies to detect peanut proteins.MethodsNon-processed and processed samples are extracted by an easy procedure at 60°C within 10 min. The measurement range is between 0.75 and 6 mg/kg peanut using a national institute of standards and technology (NIST) reference material as calibrator.ResultsThe system shows no cross-reactivity against 91 different food commodities. The LOD was 0.15 mg/kg for food matrixes such as cookies, milk chocolate, ice cream, trail mix, puffed rice cereal, and granola bar. LOQ was verified at a level of 0.75 mg/kg. Recovery studies with incurred milk chocolate and ice cream revealed consistent recoveries between 67 and 85%. Mean recoveries for incurred cookies depend on the baking temperature and time and ranged from 60 to 109%. Repeatability was between 5.2 and 12.3%, whereas relative intermediate precision was between 6.4 and 13.0%. The results for incurred cookies and milk chocolate in the independent laboratory study showed mean recoveries between 99 and 104% with RSDs between 3.56 and 19.5% under repeatability conditions.ConclusionThe results from the in-house validation study and the independent lab confirmed that the method is accurate and in accordance with requirements laid down in Standard Method Performance Requirement 2017.020.HighlightsRIDASCREEN® Peanut quantifies proteins from peanut in a wide range of food categories.