AbstractA graphene oxide (GO)-based cost-effective, automatted strip test has developed for screening of inhibitors of endonuclease EcoRV. The method involves the use of GO and a DNA substrate for EcoRV that contains both an ssDNA region for binding of GO and a fluorescein amidite (FAM)-labelled dsDNA. All the components were inkjet printed on a piece of parchment paper. The ssDNA region binds to the surface of GO and anchors so that the fluorescence of FAM is quenched. The parchment paper strip is then incubated with a sample containing EcoRV which causes enzymatic hydrolysis, and dsDNA was separated from the GO. As a result, green fluorescence is generated at the reaction spot. Enzyme activity can be measured in the presence and absence of aurintricarboxy acid acting as an EcoRV inhibitor. This method excels by its need for 2–3 orders less reagents compared to the standard well plate assay. Thus, it is an efficient platform for GO-based screening of EcoRV enzyme inhibitors. Graphical abstractA graphene oxide (GO)-based endonuclease EcoRV inhibition FRET assay using inkjet printing was developed. Printing of GO along with assay reagents has a beneficial effect on the enzymatic reaction on paper. This method was successfully applied to evaluate EcoRV inhibitor activity.

中文翻译：

---

使用喷墨印刷氧化石墨烯和 FAM 标记的 DNA 的羊皮纸对外切核酸酶 EcoRV 抑制剂进行定量的 FRET 测定

摘要已开发出一种基于氧化石墨烯 (GO) 的经济高效的自动化条带测试，用于筛选内切核酸酶 EcoRV 的抑制剂。该方法涉及使用 GO 和 EcoRV 的 DNA 底物，其中包含用于结合 GO 的 ssDNA 区域和荧光素亚酰胺 (FAM) 标记的 dsDNA。所有组件都喷墨打印在一张羊皮纸上。ssDNA 区域与 GO 表面结合并锚定，从而淬灭 FAM 的荧光。然后将羊皮纸条与含有引起酶水解的 EcoRV 的样品一起孵育，然后将 dsDNA 从 GO 中分离出来。结果，在反应点处产生绿色荧光。可以在作为 EcoRV 抑制剂的金三羧酸存在和不存在的情况下测量酶活性。与标准孔板测定相比，该方法的优势在于它需要的试剂减少 2-3 个订单。因此，它是基于 GO 筛选 EcoRV 酶抑制剂的有效平台。图形摘要开发了使用喷墨打印的基于氧化石墨烯 (GO) 的核酸内切酶 EcoRV 抑制 FRET 测定。将 GO 与检测试剂一起打印对纸上的酶促反应具有有益影响。该方法已成功应用于评估 EcoRV 抑制剂的活性。