Can the direct peptide reactivity assay be used for the identification of respiratory sensitization potential of chemicals?

Abstract

Prospective identification of low molecular weight respiratory sensitizers is difficult due to the current lack of adequate test methods. The direct peptide reactivity assay (DPRA) seems to be a promising method to determine the sensitization potential of chemicals because it determines the intrinsic characteristic of sensitizers to bind to proteins. It is already applied in the field of skin sensitization, and adaptation to respiratory sensitization has started recently. This article further evaluates the ability of the DPRA to predict the respiratory sensitization potential of chemicals. In addition, the added value of applying High Performance Liquid Chromatography (HPLC)-MS and measurements after 20 minutes and 24 hours of incubation was evaluated. Eighteen respiratory sensitizers (10 haptens, 3 prehaptens, and 5 prohaptens) and 14 nonsensitizers were tested with 2-model peptides. Based on peptide depletion, a prediction model was proposed for the identification of (respiratory) sensitizers. Application of mass spectrometry and measurements at 2 time-points increased prediction accuracy of the assay by resolving discordant results. The prediction model correctly identified all haptens and prehaptens as sensitizers. The 5 prohaptens were not identified as sensitizers, most likely due to lack of metabolic activity in the DPRA. All but 1 nonsensitizer was correctly predicted. The model, therefore, shows an accuracy of 78% for the tested dataset. Unfortunately, this assay cannot be used to distinguish respiratory from skin sensitizers. To make this distinction, the DPRA needs to be combined with other test methods that are able to identify respiratory sensitizers.