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CAN MultiFlow™
Screening assay to determine Mode of Action of Genotoxicants
This non-GLP assay uses TK6 cells to screen for Clastogens, Aneugens and Non-Genotoxicants using Flow Cytometry. All types of substances, compounds and product formulations can be tested for the presence of genotoxic modes of action that may yield positive findings and inhibit entry into further product development.
CAN MultiFlow™ is a high-throughput 96-well assay using human TK6 cells. BioReliance has validated a proprietary method and design using Litron’s MultiFlow™ DNA Damage Kit. The assay uses a logistical regression model to predict the clastogenic, aneugenic, or non-genotoxic properties of test articles based on test article-induced changes of: p53, γH2AX, Phospho-Histone H3, and polyploidy. Test articles can be loaded on to plates in multiple formats allowing for customization and optimization to fit specific needs and specific compounds. Flow cytometric analysis is employed allowing for high-throughput detection.
This assay is used to predict genotoxic as well as MOA properties of test articles in one assay, which cannot be done in other screening assays. Moreover this assay provides concurrent analysis of multiple biomarkers (p53, γH2AX, Phospho-Histone H3) and thus avoids the limitations of a single assay or biomarker. A test compound can be classified as a clastogen, aneugen, or non-genotoxicant. Identification of a clastogenic MOA means that the compound is likely DNA reactive with a linear dose response and only very low or no safe exposure is possible; and thus problematic for product development. On the other hand, an aneugen MOA means that the compound is non-DNA reactive and that there may be a threshold response and a margin of safety that allows for the compound to continue in development. The third classification is non-genotoxicant which could therefore be safe for further development. The high through-put design has short turn-around times using minimal amounts of test article. The MOA information can also be used for designing GLP assays in a smarter way to avoid costly and time consuming follow-up assays.
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