A dual-signal fluorescent probe for detection of acid phosphatase.

Acid phosphatase has become a significant indicator of prognostic and medical diagnosis, and its dysfunction may lead to a series of diseases. A novel dual-signal fluorescence method for acid phosphatase detection based on europium polymer (europium-pyridine dicarboxylicacid-adenine) and pyridoxal phosphate (PLP) was proposed. PLP coordinated with europium polymer via Eu3+ and P-O bonds, and the fluorescence of europium polymer was quenched due to the photoinduced electron transfer (PET) effect between aldehyde and europium polymer. Upon addition of acid phosphatase, the PLP was transformed to phosphate (Pi) and pyridoxal (PL). The PL was released from the surface of europium polymer, and the blue emission was enhanced due to the formation of internal hemiacetal, while the fluorescence of europium polymer recovered. The blue (PL) and red emission (Eu3+) were positively correlated with acid phosphatase activity; thus the sensitive assay of acid phosphatase was effectively achieved. The two signals were applied to determine the acid phosphatase with limits of detection (LOD) of 0.04 mU/mL and 0.38 mU/mL, and the linear ranges were 0.13-5.00 mU/mL and 1.25-20.00 mU/mL, respectively. The probe can be used to trace the acid phosphatase in biological systems and holds promise for use in clinical diagnosis and early prevention.

A fluorescent and colorimetric dual-channel sensor based on acid phosphatase-triggered blocking of internal filtration effect.

A colorimetric and fluorescent dual-channel detection method for acid phosphatase (ACP) activity has been constructed, based on the internal filtering effect between oxidized 3,3',5,5'-tetramethylbenzidine (oxTMB) and rhodamine B (RB). Au3+, which in situ form gold nanoparticles (AuNPs), can oxidize colorless 3,3',5,5'-tetramethylbenzidine (TMB) to oxTMB (blue color). The fluorescence of RB can be quenched by oxTMB due to the spectral overlap of emission of RB and absorption of oxTMB. By means of the above process, ACP can be determined because ACP promotes the hydrolysis of 2-phospho-L-ascorbic acid trisodium salt (AAP) to generate ascorbic acid (AA), which can inhibit the internal filtering effect between RB and oxTMB. No material preparation was needed for the determination of ACP. The colorimetric and fluorimetric methods can quantify ACP in the range 0.06-5.0 mU/mL and 0.03-5.0 mU/mL, respectively. Furthermore, a smartphone-assisted sensing platform has been constructed for on-site monitoring of ACP in the range 0.75-50 mU/mL, and the detection limit is 0.3 mU/mL. The methods developed can measure ACP in human serum successfully.

One-step self-assembly of magnetic supramolecular metal-organic coordination functionalized MoS2 complex as nanoenzyme-reactor.

In the present study, a kind of magnetic supramolecular metal-organic coordination complex (SMOCC) functionalized MoS2 was prepared with one-step in aqueous solution for enzyme immobilization. As possessing a protective nanocoating of PDA/PEI/Cu2+ (polydopamine: PDA, polyethyleneimine: PEI), the proposed material can provide biocompatible microenvironment and flexible adhesion force on particle interface, which is conductive to loading laccase (170.0 ± 1.8 mg/g) with high activity (93.0 ± 1.1 %). Compared with the free laccase, the immobilized laccase has higher stability in a broader range of pH (3-10), temperature (20-80 °C), storage time (1-18 days) and reusability (1-16 cycles). The removal of carcinogenic persistent organic pollutant malachite green in the water with the immobilized laccase shows a higher efficiency (89.4 ± 1.2 %) than free laccase (16.2 ± 0.2 %). The Fe3O4@MoS2@(PDA/PEI/Cu2+) nanocomposites can also be used successfully to immobilize trypsin, lipase and catalase respectively, showing a satisfactory enzyme loading (157.0 ± 0.1 mg/g, 151.6 ± 1.4 mg/g, 162.6 ± 1.6 mg/g, respectively) and activity (95.0 ± 0.5 %, 90.0 ± 0.8 %, 91.0 ± 0.9 %, respectively). The MoS2 can be replaced by carbon material and similar results can be obtained.

Ratiometric fluorescent detection and imaging of microRNA in living cells with manganese dioxide nanosheet-active DNAzyme.

MicroRNAs (miRNAs) play an important role in multiple biological processes and can be used as biomarkers for clinical disease diagnosis, so their detection is of great importance. Here, manganese dioxide (MnO2) nanosheet acts as carrier to deliver DNAzyme probes into cells through endocytosis, where intracellular glutathione (GSH) reduces the MnO2 nanosheet to manganese ions (Mn2+) and releases the probes. The generated Mn2+ can be further used as an effective cofactor to activate the DNAzyme probe, and cleave the DNA strand into two fragments. Then, the miRNA-155 in the cells can hybridize with the cleaved fragment to cause the fluorescence signal change of the probe. The proposed proportional fluorescent method has been applied to the imaging of miRNA-155 in HeLa cells and HepG2 cells with the estimated detection limit (LOD) as 1.6 × 10-12 M. The new method can provide great help for cancer diagnosis and biological research related to miRNA.