Development of a fluvoxamine detection system using a Quenchbody, a novel fluorescent biosensor
- Authors
- Sasao, Ako; Takaki, Michiyo; Jeong, Hee-Jin; Yonemitsu, Kosei; Ohtsu, Yuki; Tsutsumi, Hiroshi; Furukawa, Shota; Morioka, Hiroshi; Ueda, Hiroshi; Nishitani, Yoko
- Issue Date
- Apr-2019
- Publisher
- WILEY
- Keywords
- drug screening; fluorescent biosensor; fluvoxamine; immunoassay; recombinant antibody
- Citation
- DRUG TESTING AND ANALYSIS, v.11, no.4, pp.601 - 609
- Journal Title
- DRUG TESTING AND ANALYSIS
- Volume
- 11
- Number
- 4
- Start Page
- 601
- End Page
- 609
- URI
- https://scholarworks.bwise.kr/hongik/handle/2020.sw.hongik/1806
- DOI
- 10.1002/dta.2520
- ISSN
- 1942-7603
- Abstract
- 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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Collections - College of Science and Technology > Department of Biological and Chemical Engineering > 1. Journal Articles
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