作者:Timothy L. Foley、Adam Yasgar、Christopher J. Garcia、Ajit Jadhav、Anton Simeonov、Michael D. Burkart
DOI:10.1039/c0ob00322k
日期:——
In high throughput screening (HTS) campaigns, the quality and cost of commercial reagents suitable for pilot studies often create obstacles upon scale-up to a full screen. We faced such challenges in our efforts to implement an HTS for inhibitors of the phosphopantetheinyl transferase Sfp using an assay that had been validated using commercially available reagents. Here we demonstrate a facile route to the synthetic preparation of reactive tetraethylrhodamine and quencher probes, and their application to economically produce fluorescent and quencher-modified substrates. These probes were prepared on a scale that would allow a full, quantitative HTS of more than 350,000 compounds.
在高通量筛选(HTS)活动中,适用于初步研究的商业试剂的质量和成本往往在扩大到全面筛选时形成障碍。我们在实施针对磷酸泛妥西转移酶Sfp抑制剂的HTS时,就面临着这样的挑战,该筛选所用的检测方法已通过商业可用试剂的验证。在此,我们展示了一种简便的合成方法,用于制备反应性四乙基罗丹明和淬灭剂探针,并将其应用于经济地生产荧光标记和淬灭剂修饰的底物。这些探针的制备规模足以支持对超过350,000个化合物的全面定量HTS。