Rapid and Wash-Free Time-Gated FRET Histamine Assays Using Antibodies and Aptamers

Abstract

Histamine (HA) is an indicator of food freshness and quality. However, high concentrations of HA can cause food poisoning. Simple, rapid, sensitive, and specific quantification can enable efficient screening of HA in food and beverages. However, conventional assays are complicated and time-consuming, as they require multiple incubation, washing, and separation steps. Here, we demonstrate that time-gated Förster resonance energy transfer (TG-FRET) between terbium (Tb) complexes and organic dyes can be implemented in both immunosensors and aptasensors for simple HA quantification using a rapid, single-step, mix-and-measure assay format. Both biosensors could quantify HA at concentrations relevant in food poisoning with limits of detection of 0.19 μg/mL and 0.03 μg/mL, respectively. Excellent specificity was documented against the structurally similar food components tryptamine and l-histidine. Direct applicability of the TG-FRET assays was demonstrated by quantifying HA in spiked fish and wine samples with both excellent concentration recovery and agreement with conventional multistep enzyme-linked immunosorbent assays (ELISAs). Our results show that the simplicity and rapidity of TG-FRET assays do not compromise sensitivity, specificity, and reliability, and both immunosensors and aptasensors have a strong potential for their implementation in advanced food safety screening.

Controllable Aggregation-Induced Emission and Förster Resonance Energy Transfer Behaviors of Bistable [c2] Daisy Chain Rotaxanes for White-Light Emission and Temperature-Sensing Applications

“…Since then, AIE luminogens (AIEgens) have obtained in-depth attention and shown wide applications in light emissions, chemosensors, biosensors, and so on. − Förster resonance energy transfer (FRET) processes are well-known photophysical properties that perform the energy transfer between donor and acceptor moieties by means of nonradiative dipole–dipole couplings . Taking advantage of FRET techniques, chemists have explored different combinations of donors and acceptors in various platforms, such as small molecules, dendrimers, organo gels, polymers, biomaterials, metal–organic framework motifs, and supramolecules. − In fact, AIE properties can be incorporated with FRET processes to serve as simple and effective approaches for the development of tunable and multiple fluorescent materials . By tuning environments, AIEgens undergo the restrictions of intramolecular motions and variations of conformational structures, enabling significant changes in various excited states, so the FRET behaviors can be adjusted through the differentiation of AIE donors and/or acceptors subsequently.…”

Recent Advances in the Development and Characterization of Electrochemical and Electrical Biosensors for Small Molecule Neurotransmitters

“…Sample and reference cells are kept at the same temperature while a small amount of one partner in the interaction is repeatedly injected into the sample cell containing the other interaction partner. The required flow of heat into or out of the sample cell relative to the reference cell to maintain a steady temperature can be used to find the change in enthalpy and free energy as well as the dissociation constant for the interaction. ,,, This technique is highly sensitive but requires very accurate concentrations of the interactants. Micro-ITC instruments are also now commercially available, which allow for much lower sample consumption and required concentrations than traditional ITC.…”

Lanthanide‐FRET Molecular Beacons for microRNA Biosensing, Logic Operations, and Physical Unclonable Functions

“…[23] Many lanthanide-based TG-FRET assays have been developed, [4] including commercial tests, such as LANCE, HTRF, THUNDER, TRACE, or TruPoint. [24][25][26][27][28][29] One recent focus of TG-FRET has been its implementation into DNA biosensing, including the application of DNA aptamers for histamine quantification, [30] DNA-DNA hybridization for multiplexed sensing and the development of molecular logic devices, [19,[31][32][33] or DNA amplification for the quantification of extremely low concentrations of DNA and RNA targets. [34][35][36][37][38][39] Molecular beacons (MBs) are specific DNA FRET probes that form hairpin structures via intramolecular base-pairing, such that the 3' and 5' termini of the DNA are adjacent.…”