Shum David, Radu Constantin, Kim Earl, Cajuste Muriel, Shao Yufang, Seshan Venkatraman E, Djaballah Hakim
High Throughput Screening Core Facility, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, NY 10021, USA.
J Enzyme Inhib Med Chem. 2008 Dec;23(6):931-45. doi: 10.1080/14756360701810082.
In response to the need for inexpensive high throughput assays for anti-cancer drug screening, a 1536-well microtiter plate based assay utilizing the Alamar Blue fluorescent dye as a measure of cellular growth was validated in 10 microL assay volume. Its robustness was assessed in a screen against a library of 2000 known bioactives; with an overall Z' value of 0.89 for assay robustness, several known cytotoxic agents were identified including and not limited to anthracyclines, cardiac glycosides, gamboges, and quinones. To further test the sensitivity of the assay, IC50 determinations were performed in both 384-well and 1536-well formats and the obtained results show a very good correlation between the two density formats. These findings demonstrate that this newly developed assay is simple to set up, robust, highly sensitive and inexpensive. It could potentially provide a rapid way to screen established and primary tumor cell lines against large chemical libraries.
为满足对抗癌药物筛选的廉价高通量检测的需求，一种以1536孔微量滴定板为基础、利用阿拉玛蓝荧光染料作为细胞生长指标的检测方法在10微升检测体积下得到验证。在针对2000种已知生物活性物质的文库筛选中评估了其稳健性；该检测方法稳健性的总体Z'值为0.89，鉴定出了几种已知的细胞毒性剂，包括但不限于蒽环类药物、强心苷、藤黄和醌类。为进一步测试该检测方法的灵敏度，以384孔和1536孔两种形式进行了半数抑制浓度（IC50）测定，所得结果表明两种密度形式之间具有很好的相关性。这些发现表明，这种新开发的检测方法易于设置、稳健、高度灵敏且成本低廉。它有可能为针对大型化学文库筛选已建立的和原发性肿瘤细胞系提供一种快速方法。
