Department of Chemistry and Biochemistry, University of California, Los Angeles, Los Angeles, California 90095, United States.
California NanoSystems Institute, University of California, Los Angeles, Los Angeles, California 90095, United States.
ACS Appl Mater Interfaces. 2021 Mar 3;13(8):9425-9435. doi: 10.1021/acsami.0c17535. Epub 2021 Jan 7.
Oligonucleotide receptors (aptamers), which change conformation upon target recognition, enable electronic biosensing under high ionic-strength conditions when coupled to field-effect transistors (FETs). Because highly negatively charged aptamer backbones are influenced by ion content and concentration, biosensor performance and target sensitivities were evaluated under application conditions. For a recently identified dopamine aptamer, physiological concentrations of Mg and Ca in artificial cerebrospinal fluid produced marked potentiation of dopamine FET-sensor responses. By comparison, divalent cation-associated signal amplification was not observed for FET sensors functionalized with a recently identified serotonin aptamer or a previously reported dopamine aptamer. Circular dichroism spectroscopy revealed Mg- and Ca-induced changes in target-associated secondary structure for the new dopamine aptamer, but not the serotonin aptamer nor the old dopamine aptamer. Thioflavin T displacement corroborated the Mg dependence of the new dopamine aptamer for target detection. These findings imply allosteric binding interactions between divalent cations and dopamine for the new dopamine aptamer. Developing and testing sensors in ionic environments that reflect intended applications are best practices for identifying aptamer candidates with favorable attributes and elucidating sensing mechanisms.
寡核苷酸受体(适体)在与场效应晶体管 (FET) 结合后,在高离子强度条件下,通过构象变化来实现电子生物传感。由于高度带负电荷的适体骨架受离子含量和浓度的影响,因此在应用条件下评估了生物传感器性能和目标灵敏度。对于最近鉴定的多巴胺适体,人工脑脊液中生理浓度的 Mg 和 Ca 会显著增强多巴胺 FET 传感器的响应。相比之下,与最近鉴定的血清素适体或之前报道的多巴胺适体功能化的 FET 传感器未观察到二价阳离子相关的信号放大。圆二色光谱揭示了新的多巴胺适体与靶标相关的二级结构在 Mg 和 Ca 诱导下发生了变化,而血清素适体和旧的多巴胺适体则没有。硫代黄素 T 置换实验证实了新的多巴胺适体对靶标的检测具有 Mg 依赖性。这些发现表明,新的多巴胺适体与二价阳离子之间存在变构结合相互作用。在反映预期应用的离子环境中开发和测试传感器是识别具有有利属性的适体候选物和阐明传感机制的最佳实践。