Synthesis, characterization, and application of gold nanostars for the identification of 2-Arachidonoyl glycerol

Fereshteh Kohansal, Ahmad Mobed, M. Hasanzadeh, A. Ahmadalipour, N. Shadjou
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Abstract

Background. The endocannabinoid system (ECS) is a complex of neurotransmitters in the central nervous system (CNS) and plays a key role in regulating cognitive and physiological processes. Also, 2-Arachidonoylglycerol (2-AG) is one of the imperative endocannabinoids with key roles in the CNS. It acts as a signaling lipid and activates the cannabinoid CB1 receptor. In addition, 2-AG is involved in a variety of physiological functions such as energy balance, emotion, pain sensation, cognition, and neuroinflammation. Hence, rapid and specific diagnosis of 2-AG is of great importance in medical neuroscience. Routine methods used in detection of 2-AG are not sensitive and specific. Therefore, the development of new methods in this area has been one of the most important research areas in recent years. Methods. Herein, an innovative immunosensor was developed for quantification of 2-AG. To this end, gold nanostars (GNSs) were synthesized and conjugated with a specific biotinylated antibody activated by EDC/NHS against 2-AG. The resultant bio-conjugate was immobilized on the gold electrode surface and used to detect and analyze 2-arachidonylglycerol antigen by cyclic voltammetry (CV), square wave voltammetry (SWV), differential pulse voltammetry (DPV), and electrochemical impedance spectroscopy techniques. Results. For the first time, 2-AG protein was measured with an excellent linear range of 0.00048-1 ng/L and lower limit of quantification of 0.00048 ng/L by electroanalysis method. The designed immunosensor showed high sensitivity and specificity in the presence of interfering antigens, which proved its appropriate use in the diagnosis of neurological disorders. Conclusion. The remarkable achievement of the present study is the development of a new immunosensor for the sensitive and accurate detection of 2-AG. The fabricated sensor substrate showed good analytical results for the target analyte. Regardless of the appropriate sensitivity and selectivity of the designed biosensor, the stability of the sensor is low and it is recommended to use it immediately after preparation. Also, the immunosensor engineered in this research was very economical and had a very fast and good performance compared to the usual and traditional methods for detecting 2-AG. Practical Implications. For the first time, 2-AG protein was measured with an excellent linear range of 0.00048-1 ng/L and lower limit of quantification of 0.00048 ng/L by electro analysis method. The designed immunosensor showed high sensitivity and specificity in the presence of interfering antigens, which proved its appropriate use in the diagnosis of neurological disorders.
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金纳米星的合成、表征及在鉴定2-花生四烯醇甘油中的应用
背景。内源性大麻素系统(ECS)是中枢神经系统(CNS)中的一种神经递质复合物,在调节认知和生理过程中起关键作用。此外,2-花生四烯醇甘油(2-AG)是一种重要的内源性大麻素,在中枢神经系统中起关键作用。它作为一种信号脂质,激活大麻素CB1受体。此外,2-AG还参与能量平衡、情绪、痛觉、认知、神经炎症等多种生理功能。因此,快速、特异诊断2-AG在医学神经科学中具有重要意义。常规方法检测2-AG不敏感,特异性差。因此,在这一领域开发新方法已成为近年来最重要的研究领域之一。方法。本文开发了一种用于定量2-AG的创新免疫传感器。为此,我们合成了金纳米星(GNSs),并将其与EDC/NHS激活的特异性生物素化抗体偶联,以对抗2-AG。将合成的生物偶联物固定在金电极表面,利用循环伏安法(CV)、方波伏安法(SWV)、差分脉冲伏安法(DPV)和电化学阻抗谱技术对2-花生四烯基甘油抗原进行检测和分析。结果。2-AG蛋白首次在0.00048 ~ 1 ng/L的良好线性范围内通过电分析法测定,定量下限为0.00048 ng/L。所设计的免疫传感器对干扰抗原具有较高的敏感性和特异性,可用于神经系统疾病的诊断。结论。本研究的显著成果是开发了一种灵敏、准确检测2-AG的新型免疫传感器。所制备的传感器衬底对目标分析物具有良好的分析效果。无论所设计的生物传感器的灵敏度和选择性是否合适,传感器的稳定性都很低,建议在制备后立即使用。此外,与传统的检测2-AG的方法相比,本研究设计的免疫传感器具有非常经济、快速和良好的性能。实际意义。2-AG蛋白首次在0.00048 ~ 1 ng/L的良好线性范围内通过电分析法测定,定量下限为0.00048 ng/L。所设计的免疫传感器对干扰抗原具有较高的敏感性和特异性,可用于神经系统疾病的诊断。
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