Development of ultrafast colorimetric microplate assay method for ubiquinone utilizing the redox cycle of the quinone

Fukuda, Mizuho; Liu, Qianjun; Kishikawa, Naoya; Ohyama, Kaname; Kuroda, Naotaka

doi:10.1016/j.microc.2019.104104

Cited by 10 publications

(7 citation statements)

References 21 publications

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“…In this project, ultrahigh-performance liquid chromatography-diode array detection-quadrupole time of flight (UHPLC-DAD-QTOF) MS was used to quantify phoenicin production; however, this technique is both expensive and time-consuming compared to light absorption, which can be done on a small scale in microtiter plates with a spectrophotometer. Another microtiter plate assay for the detection of quinones is a colorimetric assay that utilizes the quinone redox flow cycle (51). Using such detection strategies could enable the possibility of screening a vast number of strains under different growth conditions.…”

Section: Discussionmentioning

confidence: 99%

Phoenicin Switch: Discovering the Trigger for Radical Phoenicin Production in Multiple Wild-Type Penicillium Species

Christiansen

Isbrandt

Asferg

et al. 2022

Appl Environ Microbiol

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Section: Discussionmentioning

confidence: 99%

Phoenicin Switch: Discovering the Trigger for Radical Phoenicin Production in Multiple Wild-Type Penicillium Species

Christiansen

Isbrandt

Asferg

et al. 2022

Appl Environ Microbiol

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“…An aliquot of NaBH 4 (50 µL; 400.0 mM, in 60 mM NaOH) was added after adding INT (100 µL; 400.0 µM) and 50 µL of Biotin-DexDox from 1.0 to 200.0 nM to the microplate’s wells, respectively. After shaking for 5 s in the microplate reader, the microplate was inserted, and the absorbance was measured at 510 nm after 5 min [ 37 ].…”

Section: Methodsmentioning

confidence: 99%

“…As indicated in Scheme 1 , the antibody-labeled quinone generates a superoxide anion radical when it reacts with a reductant such as dithiothreitol (DTT) via the redox cycle [ 32 ]. Additionally, quinone labels can react with sodium borohydride and INT, as shown in Scheme 2 for colorimetric assays [ 37 ]. The antibody can be determined by detecting this superoxide using luminol or tetrazolium dyes.…”

Section: Introductionmentioning

confidence: 99%

Anthracycline-Functionalized Dextran as a New Signal Multiplication Tagging Approach for Immunoassay

et al. 2023

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The most used kind of immunoassay is enzyme-linked immunosorbent assay (ELISA); however, enzymes suffer from steric effects, low stability, and high cost. Our research group has been developing quinone-linked immunosorbent assay (QuLISA) as a new promising approach for stable and cost-efficient immunoassay. However, the developed QuLISA suffered from low water-solubility of synthesized quinone labels and their moderate sensitivity. Herein, we developed a new approach for signal multiplication of QuLISA utilizing the water-soluble quinone anthracycline, doxorubicin, coupled with dextran for signal multiplication. A new compound, Biotin-DexDox, was prepared in which doxorubicin was assembled on oxidized dextran 40, and then it was biotinylated. The redox-cycle-based chemiluminescence and the colorimetric reaction of Biotin-DexDox were optimized and evaluated, and they showed very good sensitivity down to 0.25 and 0.23 nM, respectively. Then, Biotin-DexDox was employed for the detection of biotinylated antibodies utilizing avidin as a binder and a colorimetric assay of the formed complex through its contained doxorubicin redox reaction with NaBH4 and imidazolium salt yielding strong absorbance at 510 nm. The method could detect the plate-fixed antibody down to 0.55 nM. Hence, the application of Biotin-DexDox in QuLISA was successfully demonstrated and showed a significant improvement in its sensitivity and applicability to aqueous assays.

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“…[22][23][24] The semiquinone radicals convert dissolved oxygen to the superoxide anion radicals that can react with luminol to produce intense CL. 25,26) Although the HPLC-CL method allows sensitive detection of PQQ, there is a limitation that the CL detector is a specific device and is not widely distributed.…”

Section: Introductionmentioning

confidence: 99%

Determination Method for Pyrroloquinoline Quinone in Food Products by HPLC-UV Detection Using a Redox-Based Colorimetric Reaction

Fukuda

Kishikawa

Samemoto

et al. 2022

CHEMICAL & PHARMACEUTICAL BULLETIN

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We have developed an HPLC-UV method for the determination of pyrroloquinoline quinone (PQQ), which utilizes a redox-based colorimetric reaction. In the proposed colorimetric reaction, the redox reaction between PQQ and dithiothreitol generates superoxide anion radicals that can convert 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-phenyl-2H-tetrazolium chloride (INT) to formazan dye. After PQQ separation on an octadecyl silica column, it was mixed online with dithiothreitol and INT, and the formed formazan dye was monitored by absorbance at 490 nm. The detection limit (S/N 3) of the proposed method was 7.6 nM (152 fmol/injection). The proposed method could selectively detect PQQ in food products without any clean-up procedures.

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Development of ultrafast colorimetric microplate assay method for ubiquinone utilizing the redox cycle of the quinone

Cited by 10 publications

References 21 publications

Phoenicin Switch: Discovering the Trigger for Radical Phoenicin Production in Multiple Wild-Type Penicillium Species

Phoenicin Switch: Discovering the Trigger for Radical Phoenicin Production in Multiple Wild-Type Penicillium Species

Anthracycline-Functionalized Dextran as a New Signal Multiplication Tagging Approach for Immunoassay

Determination Method for Pyrroloquinoline Quinone in Food Products by HPLC-UV Detection Using a Redox-Based Colorimetric Reaction

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