Uni. Grenoble Alpes, CEA, CNRS, INAC-SyMMES, 17 Rue des Martyrs, 38000 Grenoble, France.
INRA, CNRS, Université de Bourgogne - Franche Comté, Centre des Sciences du Goût et de l'Alimentation, 17 rue Sully, 21000 Dijon, France.
Biosens Bioelectron. 2019 Jan 1;123:230-236. doi: 10.1016/j.bios.2018.08.072. Epub 2018 Aug 31.
Nowadays, monitoring of volatile organic compounds (VOCs) is very important in various domains. In this work, we aimed to develop sensitive olfactory biosensors using odorant binding proteins (OBPs) as sensing materials. Three rat OBP3 derivatives with customized binding properties were designed and immobilized on the same chip for the detection of VOCs in solution by surface plasmon resonance imaging (SPRi). We demonstrated that the proteins kept their binding properties after the immobilization under optimized conditions. The obtained olfactory biosensors exhibited very low limits of detection in both concentration (200 pM of β-ionone) and in molecular weight of VOCs (100 g/mol for hexanal). Such a performance obtained with SPRi in solution is especially remarkable. We hypothesized that the binding of VOCs to the active sites of OBPs induced a local conformational change in the proteins. This change would give rise to a variation of refractive index, to which SPRi is extremely sensitive. In addition, the olfactory biosensors showed a high selectivity especially at relatively low VOC concentrations. With optimized regeneration procedures, they also showed very good repeatability not only from measurement to measurement but also from chip to chip with a lifespan up to almost two months. These olfactory biosensors are particularly interesting for trace detection of VOCs in solution.
目前,挥发性有机化合物(VOCs)的监测在各个领域都非常重要。在这项工作中,我们旨在开发使用气味结合蛋白(OBP)作为传感材料的灵敏嗅觉生物传感器。设计了三种具有定制结合特性的大鼠 OBP3 衍生物,并将其固定在同一芯片上,通过表面等离子体共振成像(SPRi)检测溶液中的 VOCs。我们证明了在优化条件下固定化后,蛋白质保持其结合特性。所获得的嗅觉生物传感器在浓度(β-依兰油精 200pM)和 VOC 分子量(正己醛 100g/mol)方面均表现出非常低的检测限。在溶液中通过 SPRi 获得的这种性能尤其显著。我们假设 VOC 与 OBPs 的活性位点结合会导致蛋白质发生局部构象变化。这种变化会导致折射率发生变化,而 SPRi 对折射率的变化非常敏感。此外,嗅觉生物传感器在相对较低的 VOC 浓度下表现出很高的选择性。通过优化的再生程序,它们不仅在测量之间,而且在芯片之间也表现出非常好的重复性,使用寿命长达近两个月。这些嗅觉生物传感器特别适用于溶液中 VOC 的痕量检测。