Project Description

OECD 442D: KeratinoSensTM


The KeratinoSensTM assay is used to assess the second key event of the skin sensitisation adverse outcome pathway – activation of keratinocytes.  It does this by measuring the expression of a luciferase gene reporter gene for the ARE-dependent pathway.

Gentronix have validated their KeratinoSensTM protocols to demonstrate proficiency for the OECD guideline, and now conduct these routinely to GLP and the OECD guideline 442D.

OECD GuidelineOECD 442D
Adverse Outcome Pathway Key EventNumber 2 – activation of keratinocytes
What it measuresActivation of the Keap1-Nrf2-antioxidant/electrophile response element (ARE)-dependent pathway – using a luciferase gene reporter assay.
What is the relevance of that?Covalent binding of a substance to skin proteins is considered the first step in skin allergenicity
Cell lineKeratinoSensTM – modified HaCaT human keratinocytes
Incubation time with test item48 hours
Can be conducted to GLP?Yes
Protocol performedEURL ECVAM DB-ALM Protocol No 154

KeratinoSensTM OECD 442D Proficiency Data

  Gentronix results within OECD 442D reference range?Gentronix Results
Test chemicalIn vivo classificationEC1.5 IC50KeratinoSensTM classificationClassification correct
2,4-DinitrochlorobenzeneSensitiser (extreme)Sensitiser
4-methylamino sulfateSensitiser (strong)Sensitiser
Methyldibromo glutaronitrileSensitiser (strong)Sensitiser
2-mercapto benzothiazoleSensitiser (moderate)Sensitiser
Ethylene glycol dimethacrylateSensitiser (weak)Sensitiser
Cinnamyl alcoholSensitiser (weak)Sensitiser
Lactic acidNon-sensitiserNon-sensitiser
Salicylic acidNon-sensitiserNon-sensitiser

Predictive Toxicology, Expertly Delivered