Toward an Aptasensor for Monitoring of Tacrolimus

Bang Hyun Lee, A. Aroche, S. Menegatti, Michael A. Daniele
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Abstract

Developing a simple and accurate method for monitoring the levels of circulating tacrolimus (TAC) can inform dosing schedule and reduce the risk of rejection in solid organ transplantation. Thus, an aptasensor was designed, fabricated, and characterized for electrochemical quantification of TAC concentration. The TAC-aptasensor comprised a gold electrode functionalized with a thiolated TAC-binding aptamer (APT122) derivatized with methylene blue. Surface plasmon resonance (SPR) was used to characterize the binding strength and specificity of the TAC:APT122 complex. Square wave voltammetry (SWV) was used to quantify the aptasensor response to varying concentrations of TAC. A dose-response curve was observed at concentrations of TAC < 7 µM, when operated at 50 and 80 Hz. Typical sensitivity of 0.07 µA· µM-1 was observed at 50 Hz. Limited specificity for the TAC aptasensors was observed with SWV against cyclosporine, amoxicillin, and tetracycline. Future efforts will explore a multi-aptamer system to extend the dynamic range and improve surface functionality to limit non-specific binding.
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一种监测他克莫司的适体传感器的研制
开发一种简单而准确的方法来监测循环他克莫司(TAC)的水平,可以为给药计划提供信息,并降低实体器官移植的排斥风险。因此,设计、制作并表征了一种用于TAC浓度电化学定量的感应传感器。tac -适配体传感器包括一个金电极,该电极被亚甲基蓝衍生的硫代tac结合适配体(APT122)功能化。利用表面等离子体共振(SPR)表征TAC:APT122复合物的结合强度和特异性。方波伏安法(SWV)用于量化感应传感器对不同浓度TAC的响应。在50hz和80hz工作时,TAC < 7µM浓度下观察到剂量-响应曲线。在50 Hz时,典型灵敏度为0.07µA·µM-1。SWV对环孢素、阿莫西林和四环素的TAC适体传感器特异性有限。未来的努力将探索一个多适体系统,以扩大动态范围和改善表面功能,以限制非特异性结合。
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