Roelant C H, Burns D A, Scheirer W
Packard Instrument B. V., Groningen, The Netherlands.
Biotechniques. 1996 May;20(5):914-7. doi: 10.2144/96205pf01.
Luciferase reporter gene assays have gained more importance because of their easy readout, high sensitivity and lack of environmental waste disposal problems. However, several obstacles remain that have prohibited a wider use and the implementation of this type of assay in high-throughput screening programs: (i) Measurements need to be carried out within an active enzyme reaction, and the assessment of such reactions are time-dependent; (ii) the signal produced has a "flash" type characteristic and therefore requires specialized equipment for measurement; and (iii) side-reactions can occur that interact with the signal readout of the assay in a non-reproducible way. These hurdles make an otherwise convenient assay principle troublesome for larger-scale screening use. We have attempted to overcome these problems by different means, leading to the development of LucLite, a stable signal homogeneous reagent system. This system allows use in a higher throughput screening capacity and enables the use of standard scintillation/luminescence instruments.
荧光素酶报告基因检测因其读数简便、灵敏度高且不存在环境废物处理问题而变得更加重要。然而，仍然存在一些障碍，这些障碍阻碍了这种检测方法在高通量筛选项目中的更广泛应用和实施：（i）测量需要在活性酶反应中进行，并且此类反应的评估具有时间依赖性；（ii）产生的信号具有“闪光”型特征，因此需要专门的测量设备；（iii）可能会发生副反应，这些副反应会以不可重复的方式与检测的信号读数相互作用。这些障碍使得原本方便的检测原理在大规模筛选应用中变得麻烦。我们试图通过不同方法克服这些问题，从而开发出了LucLite，一种稳定信号的均相试剂系统。该系统允许以更高的通量进行筛选，并能够使用标准的闪烁/发光仪器。
