A direct peptide reactivity assay using a high-throughput mass spectrometry screening platform for detection of skin sensitizers
| Autor: | Yuhong Fang, Zhengxi Wei, Dingyin Tao, Anton Simeonov, Maya L. Gosztyla, Christopher A. LeClair, Andrew J. Li, Menghang Xia, Wenwei Huang |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Ethylene glycol dimethacrylate Lysine Peptide Toxicology Mass spectrometry Tandem mass spectrometry Animal Testing Alternatives High-performance liquid chromatography Risk Assessment Article Adduct 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Tandem Mass Spectrometry Toxicity Tests Humans Cysteine Chromatography High Pressure Liquid Skin chemistry.chemical_classification Chromatography Reproducibility of Results General Medicine Allergens Reference Standards High-Throughput Screening Assays 030104 developmental biology chemistry Dermatitis Allergic Contact Peptides 030217 neurology & neurosurgery |
| Zdroj: | Toxicol Lett |
| Popis: | Chemical-peptide conjugation is the molecular initiating event in skin sensitization. The OECD test guideline uses a high-performance liquid chromatography/ultraviolet (HPLC/UV) detection method to quantify chemical-peptide conjugation in a direct peptide reactivity assay (DPRA), which measures the depletion of two synthetic peptides containing lysine or cysteine residues. To improve assay throughput, sensitivity and accuracy, an automated 384-well plate-based RapidFire solid-phase extraction (SPE) system coupled with tandem mass spectrometry (MS/MS) DPRA was developed and validated in the presence of a newly designed internal standard. Compared to the HPLC/UV-based DPRA, the automated SPE-MS/MS-based DPRA improved throughput from 16 min to 10 s per sample, and substrate peptides usage was reduced from 100 mM to 5 μM. When implementing the SPE-MS/MS-based DPRA into a high-throughput platform, we found 10 compounds that depleted lysine peptide and 24 compounds that depleted cysteine peptide (including 7 unreported chemicals from 55 compounds we tested) in a concentration-response manner. The adduct formation between cysteine and cinnamic aldehyde and ethylene glycol dimethacrylate were further analyzed using high-performance liquid chromatography time-of-flight mass spectrometry (HPLC-TOF-MS) to confirm the conjugation. Overall, the automated SPE-MS/MS-based platform is an efficient, economic, and accurate way to detect skin sensitizers. |
| Databáze: | OpenAIRE |
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