Picomolar-Level Sensing of Cannabidiol by Metal Nanoparticles Functionalized with Chemically Induced Dimerization Binders.
Picomolar-Level Sensing of Cannabidiol by Metal Nanoparticles Functionalized with Chemically Induced Dimerization Binders.
复制标题
通过化学诱导二聚化粘合剂功能化的金属纳米颗粒对大麻二酚进行皮摩尔水平传感。
DOI：
10.1101/2023.09.13.557660
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Wang,Chao
中科院分区：
文献类型：
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作者：
Ikbal,MdAshif;Kang,Shoukai;Chen,Xiahui;Gu,Liangcai;Wang,Chao
小分子的简单快速检测对于健康和环境监测至关重要。化学检测方法通常使用质谱仪或酶;前者依赖于昂贵的设备，后者仅限于那些可以作为酶底物的设备。亲和试剂（如抗体）可以靶向各种小分子分析物，但检测需要成功设计用于竞争性结合测定的化学缀合靶标或类似物。在这里，我们开发了一种可推广的方法，用于高灵敏度和特异性的小分子溶液检测，以大麻二酚（CBD）为例。我们的传感平台使用金纳米粒子（AuNP）功能化与一对化学诱导二聚化（CID）纳米抗体粘合剂（nanobinders），其中CID触发AuNP聚集和沉降在CBD的存在下。尽管两种纳米粘合剂对CBD的结合亲和力适中，（平衡解离常数KD为1.6和1.56 μM），设计了由CBD-AuNP预分析孵育、离心和电子检测（ICED）组成的方案，以证明高灵敏度（检测限为100皮摩尔），相对短的感测时间（102小时），大的动态范围（5个对数），以及足够高的特异性以区分CBD与其类似物四氢大麻酚。高传感性能实现了多价的金纳米粒子传感，ICED方案，增加了分析物浓度在一个小的测定体积，和便携式电子检测器。该传感系统易于应用于广泛的分子诊断应用。
Simple and fast detection of small molecules is critical for health and environmental monitoring. Methods for chemical detection often use mass spectrometers or enzymes; the former relies on expensive equipment, and the latter is limited to those that can act as enzyme substrates. Affinity reagents like antibodies can target a variety of small-molecule analytes, but the detection requires the successful design of chemically conjugated targets or analogs for competitive binding assays. Here, we developed a generalizable method for the highly sensitive and specific in-solution detection of small molecules, using cannabidiol (CBD) as an example. Our sensing platform uses gold nanoparticles (AuNPs) functionalized with a pair of chemically induced dimerization (CID) nanobody binders (nanobinders), where CID triggers AuNP aggregation and sedimentation in the presence of CBD. Despite moderate binding affinities of the two nanobinders to CBD (equilibrium dissociation constantsKDof ∼6 and ∼56 μM), a scheme consisting of CBD–AuNP preanalytical incubation, centrifugation, and electronic detection (ICED) was devised to demonstrate a high sensitivity (limit of detection of ∼100 picomolar) in urine and saliva, a relatively short sensing time (∼2 h), a large dynamic range (5 logs), and a sufficiently high specificity to differentiate CBD from its analog, tetrahydrocannabinol. The high sensing performance was achieved with the multivalency of AuNP sensing, the ICED scheme that increases analyte concentrations in a small assay volume, and a portable electronic detector. This sensing system is readily applicable for wide molecular diagnostic applications.