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Development of a fluvoxamine detection system using a Quenchbody, a novel fluorescent biosensor.
Sasao, Ako; Takaki, Michiyo; Jeong, Hee-Jin; Yonemitsu, Kosei; Ohtsu, Yuki; Tsutsumi, Hiroshi; Furukawa, Shota; Morioka, Hiroshi; Ueda, Hiroshi; Nishitani, Yoko.
Afiliación
  • Sasao A; Department of Forensic Medicine, Faculty of Life Sciences, Kumamoto University, Kumamoto, Japan.
  • Takaki M; Department of Forensic Medicine, Faculty of Life Sciences, Kumamoto University, Kumamoto, Japan.
  • Jeong HJ; Department of Biological and Chemical Engineering, Hongik University, Sejong-si, South Korea.
  • Yonemitsu K; Department of Forensic Medicine, Faculty of Life Sciences, Kumamoto University, Kumamoto, Japan.
  • Ohtsu Y; Department of Forensic Medicine, Faculty of Life Sciences, Kumamoto University, Kumamoto, Japan.
  • Tsutsumi H; Department of Forensic Medicine, Faculty of Life Sciences, Kumamoto University, Kumamoto, Japan.
  • Furukawa S; Department of Forensic Medicine, Faculty of Life Sciences, Kumamoto University, Kumamoto, Japan.
  • Morioka H; Department of Analytical and Biophysical Chemistry, Faculty of Life Sciences, Kumamoto University, Kumamoto, Japan.
  • Ueda H; Laboratory for Chemistry and Life Science, Tokyo Institute of Technology, Yokohama, Kanagawa, Japan.
  • Nishitani Y; Department of Forensic Medicine, Faculty of Life Sciences, Kumamoto University, Kumamoto, Japan.
Drug Test Anal ; 11(4): 601-609, 2019 Apr.
Article en En | MEDLINE | ID: mdl-30328685
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.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Rodaminas / Técnicas Biosensibles / Fluvoxamina / Inmunoconjugados / Antidepresivos de Segunda Generación / Colorantes Fluorescentes Tipo de estudio: Diagnostic_studies Límite: Humans Idioma: En Revista: Drug Test Anal Asunto de la revista: FARMACOLOGIA Año: 2019 Tipo del documento: Article País de afiliación: Japón Pais de publicación: Reino Unido
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Rodaminas / Técnicas Biosensibles / Fluvoxamina / Inmunoconjugados / Antidepresivos de Segunda Generación / Colorantes Fluorescentes Tipo de estudio: Diagnostic_studies Límite: Humans Idioma: En Revista: Drug Test Anal Asunto de la revista: FARMACOLOGIA Año: 2019 Tipo del documento: Article País de afiliación: Japón Pais de publicación: Reino Unido