Abstract
For the first time, a paper-based fluorescence resonance energy transfer (FRET) determination with cyclic AMP (cAMP)-specific phosphodiesterase 4B (PDE4B) inhibitory assay using an inkjet-printing technique is proposed. Non-fabricated parchment paper is found to constitute a unique substrate to measure fluorescent energy transfer, due to its insignificant self-absorption, and enables efficient sample interaction. Here, we report the responsive FRET signals generated on paper, upon sequentially printing reaction components on parchment paper using a conventional inkjet printer equipped with four cartridges. After printing, the energy emitted by Eu chelate was transferred by FRET to ULight molecule on paper, detected at 665 nm. In the absence of free cAMP, a maximum FRET signal was achieved on paper, while a decrease in FRET signals was recorded when free cAMP produced by PDE4B inhibitors compete with Eu-cAMP, binding with ULight-mAb. The IM50 value was determined as 2.46 × 10−13 mole for roliparm and 1.86 × 10−13 mole for roflumilast, to effectively inhibit PDE4B activity. Inkjet printing-based FRET signal determination utilizes components that are less than the femtomole range, which was four-orders less than the standard assay method. The methodology reported here constitutes an innovative approach towards the determination of FRET signals generated on paper.
Highlights
In the past few years, paper-based inkjet printing biosensor and bioanalytical tools have been extensively used for the rapid detection of biomolecule interactions
In the absence of free cyclic AMP (cAMP) maximum, the fluorescence resonance energy transfer (FRET) signal was achieved on paper, while a decrease in the FRET signal was recorded when free cAMP produced by phosphodiesterase 4B (PDE4B) inhibitors compete with Eu-cAMP, binding with ULight-monoclonal antibodies (mAb)
PDE4B is a member of the phosphodiesterase family of proteins, which plays a critical role in regulating intracellular levels of cyclic AMP28. cAMP-specific 3′,5′-cyclic phosphodiesterase 4B is a therapeutic target for the treatment of several inflammatory disorders[29]
Summary
In the past few years, paper-based inkjet printing biosensor and bioanalytical tools have been extensively used for the rapid detection of biomolecule interactions. In the absence of free cAMP maximum, the FRET signal was achieved on paper, while a decrease in the FRET signal was recorded when free cAMP produced by PDE4B inhibitors compete with Eu-cAMP, binding with ULight-mAb. Parchment papers are found to be a unique substrate to measure fluorescent energy transfer, due to their insignificant self-absorption, which facilitates efficient interaction of reaction components. Parchment papers are found to be a unique substrate to measure fluorescent energy transfer, due to their insignificant self-absorption, which facilitates efficient interaction of reaction components This new parchment paper-based enzyme-inhibitor interaction/FRET assay offers several major advantages, including low reagent consumption, cost-effectiveness, no need of a fabrication process, stability, and biodegradability. The methodology reported here constitutes an innovative approach towards the quantitative determination of FRET signals generated on paper
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
