The recently reported aptamer kissing complex (AKC) strategy has allowed for the development of a new kind of sandwich-like sensing tools. Currently AKC assays have been only applied to low molecular weight molecules and their functionality in complex matrices remains challenging. The objective of the present study broken down into two sub-aims; exploring the propensity to broaden the scope of detectable analytes and designing a more robust system for potential applications to realistic samples. An all L-configuration aptaswitch module derived from a hairpin spiegelmer specific to a larger target, i.e. the arginine-vasopressin (AVP) hormone, was elaborated. The target-induced AKC formation in presence of a specific mirror-image RNA hairpin (L-aptakiss) probe were analyzed by using fluorescence anisotropy. The mirror-image kissing complex was successfully formed when the L-AVP target bound to the engineered L-aptaswitch element. It was also established that the use of methanol as cosolvent significantly improved the assay sensitivity through the stabilization of the ternary complex. Finally, the capability of the mirror-image method to operate in 10-fold diluted, untreated human serum was illustrated. The current work revealed that the AKC concept can be expanded to a wider range of targets and converted to a L-configuration sensing platform especially suitable for bioanalysis purposes.

中文翻译：

---

用于精氨酸-加压素传感的镜像适体接吻复合物

最近报道的适体接吻复合物 (AKC) 策略允许开发一种新型的类似三明治的传感工具。目前 AKC 检测仅适用于低分子量分子，它们在复杂基质中的功能仍然具有挑战性。本研究的目标分为两个子目标；探索拓宽可检测分析物范围的倾向，并设计更强大的系统以用于实际样品的潜在应用。详细阐述了源自特定于更大目标，即精氨酸加压素 (AVP) 激素的发夹镜像异构体的全 L 构型适体开关模块。通过使用荧光各向异性分析在特定镜像 RNA 发夹 (L-aptakiss) 探针存在下靶诱导的 AKC 形成。当 L-AVP 目标与工程化的 L-aptaswitch 元件结合时，成功形成了镜像接吻复合物。还确定使用甲醇作为共溶剂通过稳定三元复合物显着提高了测定灵敏度。最后，说明了镜像方法在稀释 10 倍、未经处理的人血清中操作的能力。目前的工作表明，AKC 概念可以扩展到更广泛的目标，并转换为 L 配置传感平台，特别适用于生物分析目的。说明了镜像方法在稀释 10 倍、未经处理的人血清中操作的能力。目前的工作表明，AKC 概念可以扩展到更广泛的目标，并转换为 L 配置传感平台，特别适用于生物分析目的。说明了镜像方法在稀释 10 倍、未经处理的人血清中操作的能力。目前的工作表明，AKC 概念可以扩展到更广泛的目标，并转换为 L 配置传感平台，特别适用于生物分析目的。