A new conjugated polymer-based ratiometric combination probe was constructed for adenosine triphosphate detection by taking advantage of a multisite-binding and fluorescence resonance energy transfer strategy. The method is rapid and highly selective, which can clearly discriminate ATP from persistent interferents such as ADP, AMP, other nucleoside polyphosphates and nucleobases.

We designed and synthesized a novel oligo(thiophene ethynylene) (OTE) to investigate the antibacterial activities against Gram-positive (Staphylococcus aureus and Staphylococcus epidermidis) and Gram-negative (Ralstonia solanacearum and Escherichia coli) bacteria in vitro by photodynamic therapy (PDT). Notably, OTE presents broad-spectrum and greatly high antibacterial activities after white light irradiation at nanogram per milliliter concentrations. The half inhibitory concentrations (IC50) values obtained for S. aureus, S. epidermidis, E. coli, and R. solanacearum are 8, 13, 24, and 52 ng/mL after illumination for 30 min, respectively, which are lower than that of other PDT agents. Interestingly, OTE shows the specific and very strong dark killing capability against S. aureus at the concentration of 180 ng/mL for 30 min, which is the highest efficiency biocide against S. aureus without the need of irradiation to date. The antibacterial mechanism investigated demonstrated that reactive oxygen species or singlet-oxygen generated by OTE kills bacteria irreversibly upon white light irradiation, and OTE as a v-type oligomer exerts its toxicity directly on destroying bacterial cytoplasmic membrane in the dark. Importantly, the OTE shows no cell cytotoxicity and excellent biocompatibility. The results indicate that it is potential to provide versatile applications in the efficient control of pathogenic organisms and specific application for killing S. aureus.Keywords: antibiotics; conjugated oligomer; photodynamic therapy; photosensitizer; singlet oxygen

In this paper, we described a new DNAzyme-based fluorescent biosensor for the detection of Pb2+. In the biosensor, the bulged structure is formed between the substrate labeled with fluorescein amidite (FAM) and DNAzyme after being annealed. Ethidium bromide (EB), the DNA intercalator, then intercalates into the double-stranded DNA section. Once FAM is excited, the FRET takes place from FAM to EB, which leads to the fluorescence of FAM decreasing greatly. In the presence of Pb2+, the substrate is cleaved by DNAzyme, which breaks the bulged structure. Then EB is released and the FRET from FAM to EB is inhibited. In this case, the fluorescence of FAM increases dramatically. Thus, the Pb2+ ions can be detected by measuring the fluorescence enhancement of FAM. Under optimal conditions, the increased fluorescence intensity ratio of FAM is dependent on the lead level in the sample, and exhibits a linear response over a Pb2+ concentration range of 0–100 nM with a detection limit of 530 pM. The sensor showed high selectivity in the presence of a number of interference ions. The river water samples were also tested with satisfying results by using the new method. This sensor is highly sensitive and simple without any additional treatments, which provides a platform for other biosensors based on DNAzyme.

A label-free and fluorescence turn-on aptamer biosensor has been developed for the detection of adenosine deaminase (ADA) activity with simplicity and selectivity. Adenosine aptamer will form a tight stem-loop structure upon binding with adenosine. In the absence of ADA, only a small quantity of picagreen intercalates into the stem section of aptamer, resulting in a low fluorescence of picagreen when excited at 490 nm. Interestingly, after the addition of ADA, adenosine is hydrolyzed to inosine, and the released aptamer forms double-stranded DNA (dsDNA) with its complementary single-stranded DNAc, followed by the intercalation of picagreen to dsDNA. When the solution is excited, picagreen emits strong green fluorescence. The increased fluorescence intensity of picagreen is dependent on the concentration of ADA. The detection limit of the ADA is determined to be 2 U L−1, which is lower than ADA cutoff value (4 U L−1) in the clinical requirement and more sensitive than most of the reported methods. Compared to other previous ADA sensors, the assay is not only label-free but also a turn-on signal, and possesses properties of lower cost and simpler detection system. Furthermore, this label-free strategy is also applicable to the assay of other enzymes and screening of corresponding inhibitors.

We demonstrated a sensitive and selective adenosine deaminase (ADA) detection by modulating the fluorescence resonance energy transfer (FRET) between cationic conjugated poly(9,9-bis(6′-N,N,N-trimethylammonium) hexyl)fluorine phenylene) (PFP) and the deoxyguanosine-tailored hairpin aptamer. The hairpin aptamer was labeled with a fluorophore FAM at one end and three deoxyguanosines (Gs) at the other end as a quencher. In the absence of ADA, aptamer forms hairpin-like conformation with adenosines making close affinity of Gs and FAM, which results in the weak FRET from PFP to FAM because of FAM fluorescence being quenched by Gs via photoinduced electron transfer (PET). After addition of ADA, adenosine was hydrolyzed by ADA, followed by the release of free aptamer. In this case, FAM being far away from Gs, the strong FRET thus was obtained due to the quenching process being blocked. Therefore, the new strategy based on the FRET ratio enhancement is reasonably used to detect the ADA sensitively, combining the fluorescence signal amplification of conjugated polymers with the initiative signal decreasing by Gs. The detection limit of the ADA assay is 0.3 U/L in both buffer solution and human serum, which is more sensitive than most of those previously documented methods. Importantly, the assay is rapid, homogeneous, and simple without a complicated treating process. The ADA inhibitor, erythro-9-(2-hydroxy-3-nonyl) adenine hydrochloride (EHNA), was also studied based on this assay, and the detection limit of EHNA is 10 pM. This strategy provides a new platform for the detection of other biomolecules and enzymes.Keywords: adenosine deaminase; conjugated polymers; deoxyguanosine; fluorescence resonance energy transfer; photoinduced electron transfer

We report a new biosensor for adenosine deaminase (ADA) sensing based on water-soluble conjugated poly(9,9-bis(6′-N,N,N-trimethylammonium)hexyl)fluorine phenylene (PFP) and fluorescence resonance energy transfer technique. In this biosensor, PFP, DNAc-FI labeled with fluorescein (FAM), and ethidium bromide (EB) were used as the fluorescence energy donor, resonance gate, and the final fluorescence energy acceptor, respectively. In the absence of ADA, the adenosine aptamer forms a hairpin-like conformation with adenosine, which is far from its complementary single-stranded DNA (DNAc-FI). When PFP is excited at 380 nm, fluorescein emits strong green fluorescence via one-step FRET while EB has no fluorescence. After addition of ADA, adenosine is hydrolyzed to inosine and then double-stranded DNA (dsDNA) is formed between the aptamer and DNAc-FI, followed by EB intercalating into dsDNA. Once PFP is excited, EB will emit strong yellow fluorescence after two-step FRET from PFP to fluorescein and from fluorescein to EB. The sensitive ADA detection then is realized with a low detection limit of 0.5 U/L by measuring the FRET ratio of EB to fluorescein. Most importantly, the assay is accomplished homogeneously in 25 min without further treatments, which is much more simple and rapid than that reported in literature. Hence, this method demonstrates the sensitive, cost-effective, and rapid detection of ADA activity. It also opens an opportunity for designing promising sensors for other enzymes.

New fluorescent oligo(phenylene ethynylene)–DNA hydrogels have been prepared and used for the controllable biocidal activity driven by DNase. This study opens a new way of controllable drug release and antimicrobial regulation.

A new strategy was developed and applied in monitoring pH response and enzyme activity based on fluorescence emission red shift (FERS) of the conjugated polymer PPP-OR10 induced by the inner filter effect (IFE) of nitrobenzene derivatives. Neutral poly(p-phenylenes) functionalized with oligo(oxyethylene) side chains (PPP-OR10) was designed and synthesized by the Suzuki cross-coupling reaction. Nitrobenzene derivatives display different light absorption activities in the acidic or basic form due to adopting different electron-transition types. When environmental pH is higher than their pKa values, nitrobenzene derivatives exhibit strong absorbance around 400 nm, which is close to the maximal emission of polymer PPP-OR10. As a result, the maximal emission wavelength of PPP-OR10/nitrobenzene derivatives red shifts with the pH value increasing. Apparently, the IFE plays a very important role in this case. A new method has been designed that takes advantage of this pH-sensitive platform to sensor α-chymotrypsin (ChT) based on the IFE of p-nitroaniline, since the absorption spectrum of p-nitroaniline, the ChT-hydrolyzed product of N-benzoyl-l-tyrosine-p-nitroaniline (BTNA), overlaps with the emission spectrum of PPP-OR10. In addition, the present approach can detect α-chymotrypsin with a detection limit of 0.1 μM, which is lower than that of the corresponding absorption spectroscopy method. Furthermore, the pH response and enzyme detections can be carried out in 10% serum, which makes this new FERS-based strategy promising in applications in more complex conditions and a broader field.

Reversible logic gates were designed and prepared by modulating the logic inputs based on the original system combining new cationic polyfluorene PFPB with labelled dsDNA. The NOT, YES, and NOR logic gates were built with T4 ligase and restriction endonucleases such as HaeIII and PvuII as the inputs and the emission intensity of dyes (FAM or TAMRA) as the output. Most importantly, the logic gates YES and NOT can switch back and forth by the addition of HaeIII and T4 ligase. By a similar method, the logic gates NOR and YES also realized this recycling. This study is a promising starting point for constructing reusable systems for biosensors in the multiplex detection of nucleases.

A new fluorescent biosensor has been designed to screen α-glucosidase inhibitors (AGIs) sensitively by utilizing signal amplification effect of conjugated polymers. The fluorescence of cationic poly(fluorenylene phenylene) (PFP) was quenched in the presence of para-nitrophenyl-α-d-glucopyranoside and α-glucosidase, and turned on upon addition of AGIs. Thus, a new method was developed for AGIs screening based on the fluorescence turn-off/turn-on. The IC50 values obtained for inhibitors were compared with that reported using absorption spectroscopy. All results present the new method is more sensitive and promising in screening AGIs and inhibitors of other enzymes whose hydrolysis product is 4-nitrophenol.Keywords: biosensors; conjugated polymers; diabetes; fluorescence spectroscopy; fluorescence turn-on; α-glucosidase inhibitor;

New fluorescent oligo(phenylene ethynylene)–DNA hydrogels have been prepared and used for the controllable biocidal activity driven by DNase. This study opens a new way of controllable drug release and antimicrobial regulation.