Koehler Jeffrey, Vajjhala Surekha, Coyne Courtney, Flynn Terence, Pezzuto Marci, Williams Manda, Levine Leanna
Nanostream, Pasadena, CA 91107, USA.
Assay Drug Dev Technol. 2002 Nov;1(1 Pt 1):91-6. doi: 10.1089/154065802761001347.
A novel microfluidic device was designed and developed to miniaturize, multiplex, and automate serial dilution and three-reagent dose-response assays using submicroliter quantities of reagents. This prototype microfluidic device can be used to measure enzyme kinetics and to test a chemical lead's response to a target by fluorescent readout using common plate readers and detection systems. The prototype microfluidic system yielded serial dilution and dose-response assay data comparable to results obtained from manual titrations and reagent additions performed using a microwell plate. Enzyme kinetics were highly reproducible using these devices, although Michaelis-Menten kinetics results differed from those obtained in the microwell plate. In all cases reported here, assays performed on the microfluidic format required lower volumes of reagents compared with the microwell plate. In addition to savings in reagent consumption, the microfluidic devices and bench-top instruments offer additional advantages over conventional solutions, including a small footprint and compatibility with commercially available fluorescence detectors. Future directions for the prototype technology are discussed.
设计并开发了一种新型微流控装置，以实现使用亚微升量的试剂进行串行稀释和三试剂剂量反应测定的小型化、多重化和自动化。这种原型微流控装置可用于测量酶动力学，并通过使用普通酶标仪和检测系统进行荧光读数来测试化学先导物对靶标的反应。该原型微流控系统产生的串行稀释和剂量反应测定数据与使用微孔板进行手动滴定和试剂添加所获得的结果相当。使用这些装置，酶动力学具有高度的可重复性，尽管米氏动力学结果与在微孔板中获得的结果不同。在这里报道的所有情况下，与微孔板相比，在微流控形式上进行的测定需要更少的试剂量。除了节省试剂消耗外，微流控装置和台式仪器相对于传统解决方案还具有其他优势，包括占地面积小以及与市售荧光检测器兼容。讨论了该原型技术的未来发展方向。
