Applicability of amino acid derivative reactivity assay for prediction of skin sensitization by combining multiple alternative methods to evaluate key events

Yusuke Yamamoto, Masaharu Fujita, Sayaka Wanibuchi, Ayako Sato, Miyuki Akimoto, Yasuhiro Katsuoka, Atsushi Ono, Toshihiko Kasahara

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2 Citations (Scopus)


Amino acid derivative reactivity assay (ADRA) has previously been developed as an alternative method to direct peptide reactivity assay (DPRA) to evaluate key event 1 in skin sensitization mechanisms. However, when using alternative methods for skin sensitization, integrated approaches to testing and assessment (IATA) that combine the results of multiple tests evaluating different key events are generally required. To verify whether ADRA can be used in IATA, we replaced DPRA with ADRA in five IATA methods combining DPRA, KeratinoSens, and h-CLAT: (i) the "2 out of 3" approach, (ii) the "3 out of 3" approach, (iii) sequential testing strategy (STS), (iv) integrated testing strategy by scoring approach (ITS-SA), and (v) the "ITS by two methods approach" (ITS-2MA). The prediction accuracy of the "2 out of 3" approach using ADRA (1 mM) and ADRA (0.5 mg/mL) was 90.0% and 91.1%, respectively, for human data, and was very similar to that obtained using DPRA (91.1%). The "3 out of 3" approach also showed good predictability (83.2%) using either ADRA (1 mM) or ADRA (0.5 mg/mL) compared to DPRA. Regarding the accuracy of the prediction of sensitization intensity for the human data by the third classification, prediction accuracy using ADRA was almost the same as STS, ITS-SA, or ITS-2MA using DPRA. As a result, this study showed that ADRA can be used as a test method for key event 1 in the evaluation of skin sensitization by combining multiple alternative methods.

Original languageEnglish
Pages (from-to)585-600
Number of pages16
JournalThe Journal of toxicological sciences
Issue number9
Publication statusPublished - Jan 1 2019



  • Amino acid derivative reactivity assay (ADRA)
  • Direct peptide reactivity assay (DPRA)
  • Integrated approaches to testing and assessment (IATA)
  • Integrated testing strategy (ITS)
  • Sequential testing strategy (STS)

ASJC Scopus subject areas

  • Toxicology

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