Institute for Molecular Bioscience and Australian Institute for Bioengineering and Nanotechnology, The University of Queensland , Brisbane, QLD 4072, Australia.
Molecular Warehouse Ltd. , 40 Occam Road, Guildford GU2 7YG, U.K.
J Am Chem Soc. 2016 Aug 17;138(32):10108-11. doi: 10.1021/jacs.6b06342. Epub 2016 Aug 5.
Biosensors with direct electron output hold promise for nearly seamless integration with portable electronic devices. However, so far, they have been based on naturally occurring enzymes that significantly limit the spectrum of detectable analytes. Here, we present a novel biosensor architecture based on analyte-driven intermolecular recombination and activity reconstitution of a re-engineered component of glucometers: PQQ-glucose dehydrogenase. We demonstrate that this sensor architecture can be rapidly adopted for the detection of immunosuppressant drugs, α-amylase protein, or protease activity of thrombin and Factor Xa. The biosensors could be stored in dried form without appreciable loss of activity. We further show that ligand-induced activity of the developed biosensors could be directly monitored by chronoamperometry, enabling construction of disposable sensory electrodes. We expect that this architecture could be expanded to the detection of other biochemical activities, post-translational modifications, nucleic acids, and inorganic molecules.
具有直接电子输出的生物传感器有望与便携式电子设备实现近乎无缝的集成。然而，到目前为止，它们基于天然存在的酶，这极大地限制了可检测分析物的范围。在这里，我们提出了一种基于分析物驱动的分子间重组和再工程血糖仪组件活性重建的新型生物传感器架构：PQQ-葡萄糖脱氢酶。我们证明，这种传感器架构可以快速用于检测免疫抑制剂药物、α-淀粉酶蛋白或凝血酶和因子 Xa 的蛋白酶活性。生物传感器可以以干燥形式储存，而不会明显损失活性。我们进一步表明，可以通过计时安培法直接监测配体诱导的生物传感器的活性，从而构建一次性感应电极。我们预计这种架构可以扩展到检测其他生化活性、翻译后修饰、核酸和无机分子。
