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A direct peptide reactivity assay using a high-throughput mass spectrometry screening platform for detection of skin sensitizers.

Abstract：

: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.

journal_name

Toxicol Lett

journal_title

Toxicology letters

authors

Wei Z,Fang Y,Gosztyla ML,Li AJ,Huang W,LeClair CA,Simeonov A,Tao D,Xia M

doi

10.1016/j.toxlet.2020.12.002

subject

Has Abstract

pub_date

2021-03-01 00:00:00

pages

67-77

eissn

0378-4274

issn

1879-3169

pii

S0378-4274(20)30495-1

journal_volume

338

pub_type

杂志文章

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