Department of Pharmaceutical Sciences, University of California, Irvine, CA 92697, USA.
Department of Chemistry, University of California, Irvine, CA 92697, USA.
Sensors (Basel). 2021 Nov 11;21(22):7485. doi: 10.3390/s21227485.
Current assays for in nonhospital settings are outsourced and time-intensive, resulting in both delayed diagnosis and quarantining of infected individuals. We designed a more rapid point-of-care assay featuring a "turn-on" bioluminescent readout of a -specific protease, PPEP-1. NanoLuc, a bright and stable luciferase, was "caged" with a PPEP-1-responsive peptide tail that inhibited luminescence. Upon proteolytic cleavage, the peptide was released and NanoLuc activity was restored, providing a visible readout. The bioluminescent sensor detected PPEP-1 concentrations as low as 10 nM. Sensor uncaging was achieved within minutes, and signal was captured using a digital camera. Importantly, the sensor was also functional at ambient temperature and compatible with fecal material, suggesting that it can be readily deployed in a variety of settings.
目前，非医院环境中的检测是外包的，而且非常耗时，这导致了感染个体的诊断和隔离都被延误。我们设计了一种更快速的即时检测方法，其采用了针对 PPEP-1 的“开启”生物发光读出信号的特异性蛋白酶。NanoLuc 是一种明亮且稳定的荧光素酶，其被一个响应 PPEP-1 的肽尾巴“笼锁”，抑制了发光。一旦发生蛋白水解切割，肽就会被释放，NanoLuc 的活性得到恢复，提供可见的读出信号。这种生物发光传感器可以检测到低至 10 nM 的 PPEP-1 浓度。传感器在几分钟内完成“笼锁”释放，并且使用数码相机捕获信号。重要的是，该传感器在环境温度下也能正常工作，并且与粪便标本兼容，这表明它可以很容易地在各种环境中部署。
