Development of a fluvoxamine detection system using a Quenchbody, a novel fluorescent biosensor.
Development of a fluvoxamine detection system using a Quenchbody, a novel fluorescent biosensor.
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DOI：
10.1002/dta.2520
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发表时间：
2018-11
影响因子：
2.9
通讯作者：
Ako Sasao;Michiyo Takaki;Hee-Jin Jeong;K. Yonemitsu;Y. Ohtsu;Hiroshi Tsutsumi;Shota Furukawa;H. Morioka;H. Ueda;Y. Nishitani
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文献类型：
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作者：
Ako Sasao;Michiyo Takaki;Hee-Jin Jeong;K. Yonemitsu;Y. Ohtsu;Hiroshi Tsutsumi;Shota Furukawa;H. Morioka;H. Ueda;Y. Nishitani
滥用用于医疗的精神药物是最近全世界公共卫生问题。猝灭体（Q-body）是一种新型的荧光免疫传感器，其可以立即检测抗原而无需额外的试剂或洗涤步骤。在这里，我们描述了创建Q体的抗抑郁药氟伏沙明（FLV）的检测和确定最佳条件，以实现最高的荧光强度（FI）。我们制备了五个Q体，其中荧光团在N-或C-末端标记，并且具有不同的接头长度。荧光在几分钟内可测量，表明Q体与FLV的相互作用。添加FLV后，N末端标记的Q体的标准化FI（FI比）增加约1.5倍; C末端标记的Q体未显著增加FI。在本研究中使用的荧光染料中，罗丹明6 G标记的Q体显示出最佳的FI比率。无论接头长度或标记染料如何，N末端标记的Q体的EC 50值相似（23.2- 224 nM）。我们检查了Q体是否可以适用于血清基质而不是磷酸盐缓冲盐水。完整的血清强烈干扰Q体荧光。然而，通过过滤去除蛋白质的FLV加标血清滤液的Q体FI比显示出检测FLV水平的剂量依赖性。去蛋白不干扰Q体荧光测量，可能是高灵敏度检测血清FLV所必需的。这项研究证明了Q体探针作为开发创新免疫分析应用的工具的潜力。
The misuse of psychotropic drugs intended for medical treatment represents a recent worldwide public health concern. Quenchbody (Q-body) is a novel fluoroimmunosensor that can detect an antigen immediately without additional reagents or washing steps. Here, we describe creating Q-bodies for the detection of the antidepressant fluvoxamine (FLV) and determining optimal conditions to achieve the highest fluorescence intensity (FI). We prepared five Q-bodies with the fluorophore labeled at either the N- or C- terminus and with different linker lengths. Fluorescence was measurable within minutes, indicating the interaction of Q-bodies with FLV. The normalized FI (FI ratio) of the N-terminus labeled Q-body increased approximately 1.5-fold upon FLV addition; Q-bodies labeled at the C-terminus did not significantly increase FI. Among the fluorescence dyes used in this study, Rhodamine 6G labeled Q-body showed the best FI ratio. EC50 values of the N-terminus labeled Q-bodies were similar (23.2-224nM) regardless of linker length or labeling dye. We examined whether the Q-body could be applicable to serum matrix instead of phosphate-buffered saline. The intact serum interfered strongly with the Q-body fluorescence. However, the FI ratios of the Q-body for FLV-spiked serum filtrate, for which proteins were removed by filtration, showed a dose-dependency for detecting FLV levels. Deproteinization, which does not interfere with Q-body fluorescence measurements, is likely necessary to detect serum FLV with high sensitivity. This study demonstrates the potential of Q-body probes as a tool towards developing creative immunoassay applications.