Biothiols In Serum Research Articles

A Competitive "On-Off-Enhanced On" AIE Fluorescence Switch for Detecting Biothiols Based on Hg2+ Ions and Gold Nanoclusters.

In this study, a novel "on-off-enhanced on" approach to highly sensitive rapid sensing of biothiols was developed, based on competitive modulation of gold nanoclusters (AuNCs) and Hg2+ ions. In our approach, the AuNCs were encapsulated into a zeolite imidazole framework (ZIF) for predesigned competitive aggregation-induced luminescence (AIE) emission. To readily operate this approach, the Hg2+ ions were selected as mediators to quench the fluorescence of AuNCs. Then, due to the stronger affinities between the interactions of Hg2+ ions with -SH groups in comparison to the AuNCs with -SH groups, the quenched probe of AuNCs@ZIF-8/Hg2+ displayed enhanced fluorescence after the Hg2+ ions were competitively interacted with -SH groups. Based on enhanced fluorescence, the probe for AuNCs@ZIF-8/Hg2+ had a sensitive and specific response to trace amounts of biothiols. The developed fluorescence strategy had limit of quantification (LOQ) values of 1.0 μM and 1.5 μM for Cys and GSH molecules in serum, respectively. This competitive AIE strategy provided a new direction for developing biological probes and a promising method for quantifying trace amounts of biothiols in serum. It could promote progress in disease diagnosis.

Gold alloy-based nanozyme sensor arrays for biothiol detection.

Biothiols play an important role in living cells and are associated with many diseases. Thus, it is necessary to develop a facile, cost-effective, and convenient analytical method for the detection of biothiols. Nanozymes are functional nanomaterials with enzymatic activities. Due to their unique advantages (e.g., low cost, high stability, and multifunctionality), nanozymes have been extensively used to construct sensing systems. Previous studies demonstrated colorimetric assays for biothiol detection because they could competitively inhibit the peroxidase-like activities of nanozymes. However, few studies were able to differentiate biothiols from each other. To address these challenges, herein, we first synthesized Au alloy nanozymes with better peroxidase-like activities than gold nanoparticles (AuNPs). Then, cross-reactive sensor arrays were constructed with three alloy nanozymes. Six typical biothiols (i.e., glutathione, cysteine, dithiothreitol, mercaptoacetic acid, mercaptoethanol, and mercaptosuccinic acid) were successfully detected and discriminated by the as-prepared nanozyme sensor arrays. Moreover, the practical application of the nanozyme sensor arrays was demonstrated by discriminating biothiols in serum successfully.

A single probe-based sensor array for fingerprinting biothiols in serum and urine via surfactant modulation strategy

Enormous efforts have been paid to detection of various biothiols due to their involvements in many physiological processes. In the present work, we reported a simple fluorescent sensor array for fingerprinting four different biothiols based on the same probe. The probe was a dual-fluorophore containing pyrene-pyronine FRET pair and its fluorescence emission could be easily modulated by CTAB surfactant assemblies. The three-element sensor array was built by simply varying CTAB concentration in the sensor elements that contained the same probe. The array can produce unique fluorescence fingerprints for four different biothiols and be applied to discriminate them even in biofluids like serum and urine. This study demonstrates the surfactant modulation strategy associated with using a FRET dual-fluorophore can offer an opportunity to build simple and effective fingerprinting sensing platform for multiple biothiols.

Nanozyme Sensor Arrays for Detecting Versatile Analytes from Small Molecules to Proteins and Cells.

Nanozymes have emerged as promising alternatives to overcome the high cost and low stability of natural enzymes. Nanozymes with peroxidase-like activities have been studied to construct versatile biosensors by using specific biorecognition ligands (such as enzymes, antibodies, and aptamers) or molecularly imprinted polymers (MIPs). However, the use of bioligands compromises the high stability and low cost promise of nanozymes, while the MIPs may not be applicable to multiplex detection. To address these limitations, here we constructed the nanozyme sensor arrays based on peroxidase-like Pt, Ru, and Ir nanozymes. The cross-reactive nanozyme sensor arrays were successfully used for the detection of biothiols and proteins as well as the discrimination of cancer cells because of the differential nonspecific interactions between the components of the sensor arrays and the analytes. The usefulness of the nanozyme sensor arrays was further validated by the detection of blind unknown samples, where 28 of 30 biothiols and 42 of 45 proteins were correctly identified. Moreover, the practical application of the nanozyme sensor arrays was demonstrated by the successful discrimination of biothiols in serum and proteins in human urine.

Cyclodextrin supramolecular inclusion-enhanced pyrene excimer switching for time-resolved fluorescence detection of biothiols in serum

We report here an efficient pyrene excimer signaling-based time-resolved fluorescent sensor for the measurement of biothiols (cysteine (Cys), homocysteine (Hcy), glutathione (GSH)) in human serum based on thymine–Hg2+–thymine (T–Hg2+–T) coordination chemistry and the inclusion interaction of cyclodextrin. The sensing mechanism of the approach is based on the competitive ligation of Hg2+ ions by Hcy/Cys/GSH and T–T mismatches in a bis-pyrene-labeled DNA strand with the self-complementary 5′ and 3′ ends. The introduction of γ-cyclodextrin can provide cooperation for the molecular level space proximity of the two labeled pyrene molecules, moreover the hydrophobic cavity of γ-cyclodextrin can also offer protection for the pyrene dimer's emission from the quenching effect of environmental conditions and enhance the fluorescence intensity of the pyrene excimer. When the biothiols are not presented, the sensing ensemble is in the “off” state due to the long distance between the two labeled pyrene molecules resulted from the formation of a more stable T–Hg2+–T structure. While in the presence of biothiols, Hg2+ interacts very strongly with thiol groups and the T–Hg2+–T structure is dehybridized, and then the pyrene excimer will be formed due to the self-complementary 5′ and 3′ ends of the DNA probe and the cooperation interaction of γ-cyclodextrin to pyrene dimer, thus resulting in switching the sensing ensemble to the “on” state. In the optimum conditions described, the linear concentration range of 1.0–100μM with the limit of detection (LOD) of 0.36μM for GSH was obtained. Moreover, due to the much longer lifetime of the pyrene excimer fluorescence than those of the ubiquitous endogenous fluorescent components, the time-resolved fluorescence technique has been successfully used for application in complicated biological samples.

Graphene oxide and metal-mediated base pairs based "molecular beacon" integrating with exonuclease I for fluorescence turn-on detection of biothiols.

A novel fluorescence turn-on strategy, based on the resistance of metal-mediated molecular-beacons (MBs) toward nuclease digestion and the remarkable difference in the affinity of graphene oxide (GO) with MBs and the mononucleotides, is designed for the biothiols assay. Specifically, the metal-mediated base pairs facilitate the dye labeled MBs to fold into a hairpin structure preventing the digestion by exonuclease I, and thus allow the fluorescence quenching. The competition binding by biothiols removes metal ions from the base pairs, causing the nuclease reaction, and less decrease in the fluorescence is obtained after incubating with GO due to the weak affinity of the product-mononucleotides to GO. Hg(2+)-mediated MBs were firstly designed for the biothiols detection, and glutathione (GSH) was applied as the model target. Under the optimal conditions, the approach exhibits high sensitivity to GSH with a detection limit of 1.53 nM. Ag(+)-mediated MBs based sensor was also constructed to demonstrate its versatility, and cysteine was studied as the model target. The satisfactory results in the determination of biothiols in serum demonstrate that the method possesses great potential for detecting thiols in biological fluids. This new approach is expected to promote the exploitation of metal-mediated base pairs-based biosensors in biochemical and biomedical studies.