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BIOLUMINESCENT SCREENING TEST FOR DETECTING CELL CYTOLYSIS

机译:生物发光筛选试验检测细胞的细胞溶解

摘要

The present invention relates to an in vitro method for testing the cytotoxicity of a candidate compound for living target cells, said method relying on the detection of the massive release of a cytosolic luminescent enzyme in the supernatant of the cells once they have been voluntarily permeabilized. The detection of this release is rapid and reproducible. More precisely, the method of the invention comprises the steps: a) contacting, with a candidate compound,living target cells that constitutively express in their cytosol a luminescent enzyme, b) isolating the supernatant of the coculture of step a) in a second recipient, c) adding in said second recipient the substrate of said luminescent enzyme, and d) measuring the luminescence emitted in said second recipient. The cytotoxicity of said candidate compound is then proportional to the increase in luminescence measured in step d). This method can be advantageously automated so as to assess the cytotoxicity of candidate compounds with high throughput screening devices.
机译:本发明涉及一种体外方法,用于测试候选化合物对活靶细胞的细胞毒性,所述方法依赖于检测细胞上清液中胞质发光酶的大量释放。一旦他们被自愿渗透。这种释放的检测是快速且可重复的。更准确地,本发明的方法包括以下步骤:a)使候选化合物与在其细胞质中组成性表达发光酶的活靶细胞接触,b)在第二个受体中分离步骤a)的共培养上清液; c)在所述第二接收器中添加所述发光酶的底物,和d)测量在所述第二接收器中发射的发光。然后,所述候选化合物的细胞毒性与在步骤d)中测量的发光增加成比例。该方法可以有利地自动化,以便通过高通量筛选装置评估候选化合物的细胞毒性。

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