Gentronix Launches New Genotoxic Metabolite Detection Assay
10 Mar 2008Now available with an S9 protocol to detect genotoxic metabolites, GreenScreen HC from Gentronix is a human cell-based genotoxicity screening assay used in drug discovery programmes during lead compound optimisation and candidate selection.
Potential pharmaceutical compounds which only present a genotoxicity hazard after metabolism, usually by the liver, are known as pro-genotoxins. The utility of GreenScreen HC has now been extended to detect pro-genotoxins as well as all common mechanistic classes of genotoxin including mutagens, clastogens, aneugens, as well as both topoisomerase and polymerase inhibitors.
Using a standardized 96-well microplate format, the test compound and patented Green Fluorescent Protein (GFP) reporter cells are combined with 1% S9 for a 3 hour exposure period. After exposure, cells are washed to remove the S9 and test compound and then allowed a further 45 hour recovery incubation. Cell viability and GFP fluorescence data are assessed using flow cytometry and simple software generates quantitative results and a graphical output.
Minimal investment in early genotoxicity screening delivers major time savings, avoids unnecessary animal testing and prevents costly late stage failures of lead drug compounds through early detection of genotoxic potential. Combining exceptional levels of specificity and sensitivity, GreenScreen HC also correctly identifies non-carcinogenic compounds known to give misleading positive results in other in vitro genotoxicity assays.
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GreenScreen HC
Gentronix Ltd.Gentronix has launched a new, accurate and fast in vitro mammalian cell genotoxicity assay - GreenScreen HC. The TK6 host cells are p53 competent and familiar to most genetic toxicology laboratories. A patented GFP reporter system exploits the proper regulation of the GADD45a gene. The assay delivers both high specificity and high sensitivity and detects all common mechanistic classes of genotoxin. The 96-well microplate format takes about 20 minutes to prepare and delivers results after a 48 hour incubation. A single microplate is sufficient for the simultaneous testing of 4 compounds over 9 serial dilutions. The protocol is also readily automated using standard laboratory liquid-handling equipment. The Biology GADD45a mediates the adaptive response to genotoxic stress. The patented GFP fluorescence reporter includes complex regulatory elements. The assay gives positive results for direct acting agents, as well as aneugens, and topoisomerase and polymerase inhibitors. Importantly the assay gives correct negative results for non-carcinogens, including many which give misleading positive results in other in vitro tests. The Assay Protocol Nine, 2-fold dilutions of each compound together with positive controls are set out in the microplate and growing cells are added to each well. After incubation and measurement in a microplate reader, simple software gives automated decisions and a clear graphical output. Validation A unique combination of both high-specificity and high-sensitivity genotoxicity assessment in a human cell line has been demonstrated. See: High-specificity and high-sensitivity genotoxicity assessment in a human cell line: Validation of the GreenScreen HC GADD45a-GFP genotoxicity assay. Hastwell P.W. et al (2006) Mutation Research 607: 160-175.