Mass Spectrometry-Based Solid Phase Peptide Reaction Assay for Detecting Allergenicity Using an Immobilized Peptide-Conjugating Photo-Cleavable Linker

International Journal of Molecular Sciences

Yoshio Hamada¹  Kazuya Iwata¹  Shinya Hikida¹  Yasutaka Samejima¹  Yuuki Minamino¹  Kenji Usui¹  Hideto Ariumi²  Kunihiko Yamashita³  Hiroshi Miyazaki³  Hidefumi Ikeda⁴ 

[1] Faculty of Frontiers of Innovative Research in Science and Technology (FIRST), Konan University, Chuo-ku, Kobe 650-0047, Japan;Faculty of Pharmaceutical Sciences, Sanyo-Onoda City University, Sanyo-Onoda, Yamaguchi 756-0884, Japan;Medical Device Division, Innovation and Business Development Headquarters, Daicel Corporation, Minato-ku, Tokyo 108-8230, Japan;Product Assurance Division, Mandom Corporation, Chuo-ku, Osaka 540-8530, Japan;

关键词: alternative to animal testing;    immobilization;    peptides;    skin sensitizers;    mass spectrometry;    photo-labile linker;   

DOI  :  10.3390/ijms21218332

来源: DOAJ

【 摘 要 】

The biological process of skin sensitization depends on the ability of a sensitizer to modify endogenous proteins. A direct peptide reactivity assay (DPRA), based on the biological process of skin sensitization, was developed as an alternative to controversial animal experiments. Although DPRA has been endorsed by industries and is internationally accepted as promising, it has several drawbacks, such as incompatibility with hydrophobic chemicals, inability to perform detailed reaction analysis, and ability to evaluate only single components. Here, we demonstrated that sensitizers and peptide adducts can be easily identified using a mass spectrometry-based solid-phase peptide reaction assay (M-SPRA). We synthesized peptides with a photo-cleavable linker immobilized on resins. We showed the potential of M-SPRA in predicting skin sensitization by measuring the peptide adducts that were selectively eluted from the resin after cleaving the linker post-reaction. M-SPRA provides more detailed information regarding chemical reactivity and accurate assessment of test samples, including mixtures. M-SPRA may be helpful for understanding the binding mechanism of sensitizers (toxicology), which may assist in further refining reactivity assays and aiding in the interpretation of reactivity data.

【 授权许可】

Unknown