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A pre-validation transferability study of the GreenScreen HC GADD45a-GFP assay with a metabolic activation system (S9)

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Title: A pre-validation transferability study of the GreenScreen HC GADD45a-GFP assay with a metabolic activation system (S9)
Authors: Billinton, Nicholas1 nick.billinton@gentronix.co.uk, Bruce, Shannon2 Shannon.Bruce@bioreliance.com, Hansen, Julia Rytter3 julia.hansen@leo-pharma.com, Hastwell, Paul W.4 paul.w.hastwell@gsk.com, Jagger, Christopher1 christopher.jagger@gentronix.co.uk, McComb, Catriona4 catriona.x.mccomb@gsk.com, Klug, Michelle L.2 Michelle.Klug@bioreliance.com, Pant, Kamala2 Kamala.Pant@bioreliance.com, Rabinowitz, Adam1 adamrabs@hotmail.com, Rees, Robert4 Robert.W.Rees@gsk.com, Tate, Matthew1,5 matthew.tate@gentronix.co.uk, Vinggaard, Anne Marie3 rie.vinggaard@leo-pharma.com, Walmsley, Richard M.1,5 richard.walmsley@manchester.ac.uk
Source: Mutation Research - Genetic Toxicology & Environmental Mutagenesis. Jul2010, Vol. 700 Issue 1/2, p44-50. 7p.
Subject Terms: *Genetic toxicology, *Heterocyclic compounds, Biotransformation (Metabolism), Medical protocols, Flow cytometry, Fluorescence, Reporter genes
Abstract: Abstract: A new protocol has recently been developed and validated for the GreenScreen HC GADD45a-GFP genotoxicity reporter assay, enabling the incorporation of an S9 metabolic activation system into the assay. The S9 protocol employs flow-cytometric methodology for the detection of both reporter GFP fluorescence and propidium iodide fluorescence for the estimation of cellular viability. In the spirit of assay validation by bodies such as the European Centre for the Validation of Alternative Methods (ECVAM), the adapted metabolic activation protocol for the GADD45a-GFP assay has been undergoing ‘pre-validation’. Results of phases I and II of this pre-validation, namely protocol refinement and protocol transfer, respectively, are presented here. In phase I the protocol was transferred to a second laboratory for initial assessment of method portability and subsequent refinement of the protocol. In phase II, the protocol was then transferred to two further laboratories along with the elaborated standard operating-procedure (SOP) for further assessment of transferability. The three transfer sites then undertook an assessment of the method''s reproducibility by testing eight compounds. The outcome of the study was a refined protocol that was found to be highly transferable. It yielded 100% agreement in results between all four laboratories. [Copyright &y& Elsevier]
Copyright of Mutation Research - Genetic Toxicology & Environmental Mutagenesis is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
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DbLabel: GreenFILE
An: 51848022
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PubTypeId: academicJournal
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  Data: A pre-validation transferability study of the GreenScreen HC GADD45a-GFP assay with a metabolic activation system (S9)
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  Data: <searchLink fieldCode="AR" term="%22Billinton%2C+Nicholas%22">Billinton, Nicholas</searchLink><relatesTo>1</relatesTo><i> nick.billinton@gentronix.co.uk</i><br /><searchLink fieldCode="AR" term="%22Bruce%2C+Shannon%22">Bruce, Shannon</searchLink><relatesTo>2</relatesTo><i> Shannon.Bruce@bioreliance.com</i><br /><searchLink fieldCode="AR" term="%22Hansen%2C+Julia+Rytter%22">Hansen, Julia Rytter</searchLink><relatesTo>3</relatesTo><i> julia.hansen@leo-pharma.com</i><br /><searchLink fieldCode="AR" term="%22Hastwell%2C+Paul+W%2E%22">Hastwell, Paul W.</searchLink><relatesTo>4</relatesTo><i> paul.w.hastwell@gsk.com</i><br /><searchLink fieldCode="AR" term="%22Jagger%2C+Christopher%22">Jagger, Christopher</searchLink><relatesTo>1</relatesTo><i> christopher.jagger@gentronix.co.uk</i><br /><searchLink fieldCode="AR" term="%22McComb%2C+Catriona%22">McComb, Catriona</searchLink><relatesTo>4</relatesTo><i> catriona.x.mccomb@gsk.com</i><br /><searchLink fieldCode="AR" term="%22Klug%2C+Michelle+L%2E%22">Klug, Michelle L.</searchLink><relatesTo>2</relatesTo><i> Michelle.Klug@bioreliance.com</i><br /><searchLink fieldCode="AR" term="%22Pant%2C+Kamala%22">Pant, Kamala</searchLink><relatesTo>2</relatesTo><i> Kamala.Pant@bioreliance.com</i><br /><searchLink fieldCode="AR" term="%22Rabinowitz%2C+Adam%22">Rabinowitz, Adam</searchLink><relatesTo>1</relatesTo><i> adamrabs@hotmail.com</i><br /><searchLink fieldCode="AR" term="%22Rees%2C+Robert%22">Rees, Robert</searchLink><relatesTo>4</relatesTo><i> Robert.W.Rees@gsk.com</i><br /><searchLink fieldCode="AR" term="%22Tate%2C+Matthew%22">Tate, Matthew</searchLink><relatesTo>1,5</relatesTo><i> matthew.tate@gentronix.co.uk</i><br /><searchLink fieldCode="AR" term="%22Vinggaard%2C+Anne+Marie%22">Vinggaard, Anne Marie</searchLink><relatesTo>3</relatesTo><i> rie.vinggaard@leo-pharma.com</i><br /><searchLink fieldCode="AR" term="%22Walmsley%2C+Richard+M%2E%22">Walmsley, Richard M.</searchLink><relatesTo>1,5</relatesTo><i> richard.walmsley@manchester.ac.uk</i>
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  Data: <searchLink fieldCode="JN" term="%22Mutation+Research+-+Genetic+Toxicology+%26+Environmental+Mutagenesis%22">Mutation Research - Genetic Toxicology & Environmental Mutagenesis</searchLink>. Jul2010, Vol. 700 Issue 1/2, p44-50. 7p.
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  Data: *<searchLink fieldCode="DE" term="%22Genetic+toxicology%22">Genetic toxicology</searchLink><br />*<searchLink fieldCode="DE" term="%22Heterocyclic+compounds%22">Heterocyclic compounds</searchLink><br /><searchLink fieldCode="DE" term="%22Biotransformation+%28Metabolism%29%22">Biotransformation (Metabolism)</searchLink><br /><searchLink fieldCode="DE" term="%22Medical+protocols%22">Medical protocols</searchLink><br /><searchLink fieldCode="DE" term="%22Flow+cytometry%22">Flow cytometry</searchLink><br /><searchLink fieldCode="DE" term="%22Fluorescence%22">Fluorescence</searchLink><br /><searchLink fieldCode="DE" term="%22Reporter+genes%22">Reporter genes</searchLink>
– Name: Abstract
  Label: Abstract
  Group: Ab
  Data: Abstract: A new protocol has recently been developed and validated for the GreenScreen HC GADD45a-GFP genotoxicity reporter assay, enabling the incorporation of an S9 metabolic activation system into the assay. The S9 protocol employs flow-cytometric methodology for the detection of both reporter GFP fluorescence and propidium iodide fluorescence for the estimation of cellular viability. In the spirit of assay validation by bodies such as the European Centre for the Validation of Alternative Methods (ECVAM), the adapted metabolic activation protocol for the GADD45a-GFP assay has been undergoing ‘pre-validation’. Results of phases I and II of this pre-validation, namely protocol refinement and protocol transfer, respectively, are presented here. In phase I the protocol was transferred to a second laboratory for initial assessment of method portability and subsequent refinement of the protocol. In phase II, the protocol was then transferred to two further laboratories along with the elaborated standard operating-procedure (SOP) for further assessment of transferability. The three transfer sites then undertook an assessment of the method''s reproducibility by testing eight compounds. The outcome of the study was a refined protocol that was found to be highly transferable. It yielded 100% agreement in results between all four laboratories. [Copyright &y& Elsevier]
– Name: AbstractSuppliedCopyright
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  Data: <i>Copyright of Mutation Research - Genetic Toxicology & Environmental Mutagenesis is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.)
PLink https://search.ebscohost.com/login.aspx?direct=true&site=eds-live&db=8gh&AN=51848022
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        Value: 10.1016/j.mrgentox.2010.05.001
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      – SubjectFull: Biotransformation (Metabolism)
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