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HTS assays for cytotoxic molecules

The identification of apoptotic drugs remains one of the highest priorities of the pharmaceutical industry, with over 100 products being listed in development in 2001. Most drugs in development for cancer will at some time in their development be tested for apoptotic activity and indeed HTS of proapoptotic molecules remains a strategy for identifying new cancer therapies. On the other hand, the inhibition of apoptosis is an option for the treatment of various ischemic diseases such as myocardial infarction and neuro-degeneration. In order to identify both pro- and anti-apoptotic molecules, reproducible, easy-to-handle, and rapid microplate-based assays that are amenable to HTS are required. Researchers from PROCREA BioSciences have recently developed such an HTS compatible assay based on the measurement of fluorescence of cells expressing enhanced green fluorescent protein (EGFP). Cells expressing EGFP exhibited a decrease in fluorescence when treated by various agents, such as chemotherapeutic drugs, UV irradiation, or caspase-independent cell death inducers. Kinetics and sensitivity of this EGFP-based assay were comparable to those of traditional apoptosis markers such as annexin-V binding, propidium iodide incorporation, or reactive oxygen species production. This assay may represent an improved strategy for identifying at high throughput, pro- and anti-apoptotic molecules.


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