H2AX ICW method, gH2AX In cell western

Topic: Genotoxicity/Mutagenicity

Test Method Number:

TM2014-02 (EU)

Short Name of TM:

H2AX ICW

Year received:

2014

Responsible Organisation:

EURL ECVAM - European Union

General Comments:

Presubmission assessment finalised. Method Not prioritised for validation

Method Description

The method is an in vitro genotoxic 96-well plate format assay for the measurement of histone gH2AX phosphorylation, normally occurring after DSBs induction in different cell types. The use of the gH2AX global genotoxic biomarker coupled with the in cell western technique permit to determined simultaneously the cytotoxicity and the genotoxicity of any tested compound.

The in cell western technique is an immunohistochemistry in situ (directly into the well) used in combination with infra-red fluorescence. The assay is proposed in early discovery phase of drug development for genotoxicity screening or as a substitute of the in vitro micronucleus test.

Track Approval Status

Submission
Validation
Peer-review
Recommendation
Regulatory acceptance/Standards

Submission

Show status

Date	Stage	Comment
Jun 2014	Receipt of presubmission	Presubmission(TPF)received
Jul 2014	Assessment of presubmission	Presubmission assessment finalised. A revised pre-submission requested from test submitter.
Nov 2014	Not considered for validation	In the context of the current regulatory testing strategy for genotoxicity assessment, the method would not appear to offer a clear added value. There is also a lack of mechanistic information

Validation

Peer-review

Recommendation

Regulatory acceptance/Standards