Prostate Specific Antigen Aptamer Research Articles

A self-powered photoelectrochemical aptasensor based on target response and Ag-CdIn2S4 photosensitive material for ultrasensitive detection of prostate-specific antigen

In this study, Ag-CdIn2S4 was selected as a photosensitive material and combined with the zeolite imidazole frame-90 encapsulated methylene blue (ZIF-90@MB) to establish a controlled release system. On that basis, a self-powered photoelectrochemical (PEC) aptasensor was constructed based on target responses to realize the highly sensitive detection of prostate-specific antigen (PSA). The self-powered sensing method was employed to effectively avoid interference from other oxidizing/reducing substances in the system and improve the accuracy of the detection method. The band gap of CdIn2S4 was adjusted by Ag doping, and the visible light absorption capacity and photoelectric conversion efficiency of the material were improved by the local surface plasmon resonance (LSPR) effect of metal Ag. The ZIF-90 porous material was used as a carrier, the MB photosensitizer as a signal molecule was loaded in the ZIF-90 pore channel. The PSA aptamer (PApt) was attached to the ZIF-90@MB surface to form a simple biological gate. In the presence of the PSA, the recognition interaction between PApt and PSA made the biological gate open, then many MB molecules were released from ZIF-90 and enhanced the photocurrent signal. Under the optimal experimental conditions, the linear range of the sensor was 0.005 ∼ 50 ng mL−1, and the detection limit was 2.35 pg mL−1. The accurate determination of PSA in human serum was achieved, which provided a novel sensing strategy for the early diagnosis of clinical biomarkers.

Ultrasensitive detection of PSA in human serum using Label-Free electrochemical biosensor with magnetically induced Self-Assembly based on α-Fe2O3/Fe3O4@Au nanocomposites

To achieve the goals of early diagnosis, screening, and precision medicine for prostate cancer, this research designed a label-free electrochemical biosensor by combining magnetic probes (PSAA/α-Fe2O3/Fe3O4@Au) with magnetically induced self-assembly (MISA) technology to explore the application of prostate-specific antigen (PSA) in precise detection. Magnetic α-Fe2O3/Fe3O4 nanoparticles (NPs) were constructed using a hydrothermal-calcination reduction method. Subsequently, α-Fe2O3/Fe3O4@Au nanocomposites were prepared by reducing HAuCl4 with NaBH4 at a low temperature. The self-assembly method was adopted in two steps: firstly, the sulfhydryl-modified PSA aptamer is immobilized onto the material surface using Au–S bonds; secondly, the magnetic NPs and magnetic glassy carbon electrode (MGCE) undergo self-assembly. The biosensor showed good linear response in the range of 100 fg/mL to 100 ng/mL, with a detection limit of 0.78 pg/mL (S/N = 3) under optimal conditions. Repeatability, stability, selectivity, reproducibility, and the capability to analyze real samples were also investigated. With its cost-effectiveness, operational simplicity, and speed of analysis, the biosensing system platform holds great potential as a valuable tool in clinical PSA detection for point-of-care testing (POCT).

Ratio fluorescence/colorimetric dual mode aptasensor and smartphones-assisted miniaturized device for early diagnosis of prostate cancer

Prostate specific antigen (PSA) is the most common tumor biomarker for prostate cancer (PCa), and its precise determination is essential to improving survival rate and reducing treatment cost of cancer patient. Here, we have proposed a novel ratio fluorescence/colorimetric dual mode aptasensor based on CdTe-apt + B-CDs + ZrFe MOF system for PSA determination. Specifically, as-prepared ZrFe MOF with good peroxidase like activity can quench the fluorescence of B-CDs + CdTe-apt system via inner filter effect (IFE), while the existence of PSA can restore fluorescence of CdTe-apt but virtually unchanged fluorescence of B-CDs due to specific identification between PSA and aptamer. Similarly, the addition of PSA can recover the peroxidase like activity of ZrFe MOF which is significantly suppressed by PSA aptamer. Based on this, the proposed ratio fluorescence/colorimetric dual mode aptasensor has achieved satisfactory results in PSA analysis of serum specimens. Valuably, an inexpensive, portable but credible smartphones-assisted miniaturized device is expected to be exploited for the early diagnosis of PCa, inspiring the design and construction of advanced portable devices for self-monitoring and home screening of various cancers.

Highly Efficient Signal On/Off Electrochemiluminescence Gel Aptasensor Based on a Controlled Release Strategy for the Sensitive Detection of Prostate Specific Antigen.

The controlled release strategy can make the constructed sensor have the function of self-on/off, which has an obvious effect on improving the sensitivity in immunoassays. Metal organic gels (MOGs) are the most noteworthy. They are materials with ultrahigh surface area, highly dispersed atomical metal sites, and well-defined porosity and can be used as an efficient luminophore to cause the developed sensor to have good hydrophilicity and adjustability, thus further improving the detection sensitivity. In this work, a novel on/off electrochemiluminescence (ECL) gel aptasensor was constructed using the Cys-[Ru(dcbpy)3]2+ gel as a luminophore, ZnS quantum dots (QDs) as quenchers, and aminated mesoporous silica nanocontainers (SiO2-NH2) as carriers of controlled release for prostate specific antigen (PSA) detection. Specifically, the ssDNA and PSA aptamer made up clamp-like molecules to block holes of the SiO2-NH2 after encapsulating the quencher ZnS QDs. Because of the specific binding between the PSA antigen and aptamer, the clamp-like molecules of ssDNA and the PSA aptamer were disassembled. Finally, the release of ZnS QDs was triggered, thereby realizing a self-off mode of the ECL signals under a co-reactant-free environment by ECL resonance energy transfer (ECL-RET) between the Cys-[Ru(dcbpy)3]2+ and ZnS QDs. In addition, the quenching mechanism was confirmed by molecular orbitals from the theoretical calculation level. The detection limit of the gel aptasensor for PSA was as low as 1.01 fg/mL, showing excellent sensitivity and accuracy. These strategies provided a feasible idea for PSA and even other tumor marker immunoassays.

Sensitive determination of prostate-specific antigen with graphene quantum dot-based fluorescence aptasensor using few-layer V2CTx MXene as quencher

A novel fluorescence aptasensor of prostate-specific antigen (PSA) was established using few-layer vanadium carbide (FL-V2CTx) nanosheet as a quencher. First, FL-V2CTx was prepared by the delamination of multi-layer V2CTx (ML-V2CTx) with tetramethylammonium hydroxide. The aptamer-carboxyl graphene quantum dots (CGQDs) probe was prepared by combining the aminated PSA aptamer and CGQDs. Then, the aptamer-CGQDs were absorbed onto the surface of FL-V2CTx by hydrogen bond interaction, which led to the decrease in fluorescence of aptamer-CGQDs due to photoinduced energy transfer. After addition of PSA, PSA-aptamer-CGQDs complex was released from FL-V2CTx. The fluorescence intensity of aptamer-CGQDs-FL-V2CTx with PSA was higher than that without PSA. The FL-V2CTx-based fluorescence aptasensor provided a PSA detection linear range from 0.1 to 20 ng mL−1 with detection limit of 0.03 ng mL−1. The ΔF value of fluorescence intensities for aptamer-CGQDs-FL-V2CTx with and without PSA was 5.6, 3.7, 7.7, and 5.4 times of ML-V2CTx, few-layer titanium carbide (FL-Ti3C2Tx), ML-Ti3C2Tx and graphene oxide aptasensors, respectively, indicating the advantage of FL-V2CTx. The aptasensor had high selectivity for PSA detection compared with some proteins and tumor markers. This proposed method had convenience and high sensitivity for PSA determination. The determination results of PSA in human serum samples using the aptasensor were consistent with those by chemiluminescent immunoanalysis. The fluorescence aptasensor can be successfully applied for PSA determination in serum samples of prostate cancer patients.

A novel fluorescence aptasensor based on PCN-223 as an efficient quencher for sensitive determination of prostate-specific antigen.

A novel fluorescence aptasensor based on PCN-223 as an efficient quencher was developed to sensitively detect prostate-specific antigen (PSA). The 5-carboxytetramethylrhodamine (TAMRA)-labeled PSA aptamer was adsorbed on PCN-223 by π-π stacking and hydrogen-bonding interactions, which contributed to fluorescence quenching because of the photoinduced electron transfer from TAMRA to PCN-223. In addition, the amount ofquenched fluorescence of the PSA-binding aptamer complex-PCN-223 was lower than that of TAMRA aptamer-PCN-223 without PSA (at excitation/emission peaks of 545/582nm), which can be explained by the factthat the PSA-binding aptamer complexes contributed to the separation of the aptamer from PCN-223. ∆F value of fluorescence intensities for TAMRA aptamer-PCN-223 with and without PSA showed a good linear relationship with PSA concentration over a range of 0.1 to 24ngmL-1, with a detection limit of 0.05ngmL-1. Compared with three metal-organic frameworks (MOFs) of UiO-66-NH2, ZIF-67, and Ni3(HITP)2 as quenchers, PCN-223 as a Zr-MOF exhibited the highest ∆F value for PSA detection. The advantage of PCN-223 could be attributed to its carboxyl, benzene, and porphyrin groups, the large specific surface area and good biocompatibility. This proposed aptasensor can be successfully used to detect PSA in sera of prostate cancer patients. The PSA detection results of this aptasensor were consistent with those which were obtained from hospital by Archtecti2000sr automatic chemiluminescence immunoanalyzer. The proposed aptasensor has potential clinical detection application.

Gold Nanoparticles-MWCNT Based Aptasensor for Early Diagnosis of Prostate Cancer.

Prostate cancer is one of the most frequently diagnosed male malignancies and can be detected by prostate-specific antigen (PSA) as a biomarker. To detect PSA, several studies have proposed using antibodies, which are not economical and require a long reaction time. In this study, we propose to use self-assembled thiolated single-strand DNA on electrodes functionalized by multi-walled carbon nanotubes (MWCNT) modified with gold nanoparticles (AuNPs) to realize a low-cost label-free electrochemical biosensor. In this regard, the PSA aptamer was immobilized via electrostatic adsorption on the surface of a screen-printed MWCNT/AuNPs electrode. The immobilization process was enhanced due to the presence of Au nanoparticles on the surface of the electrode. Surface characterization of the electrode at different stages of modification was performed by electrochemical impedance spectroscopy (EIS), atomic force microscopy (AFM) and Fourier transform infrared spectroscopy (FTIR) and contact angle for surface tension properties. The results showed an increase in surface roughness due to the absorbance of the aptamer on the electrode surfaces. The developed sensor has an extended linear range of 1-100 ng/mL, and a very low limit of detection down to 1 pg/mL. In addition, the reaction has a binding time of only five minutes on the developed electrodes. Investigations of the biosensor selectivity against several substances revealed an efficient selectivity for PSA detection. With this approach, low-cost biosensors with high sensitivity can be realized which have a wide linearity range and a low limit of detection, which are necessary for the early detection of prostate cancer.

Detection of prostate cancer biomarkers via a SERS-based aptasensor

The overexpression of specific biomarkers in serum is closely related to diseases, and accurate and sensitive detection of them is beneficial for the early diagnosis and treatment of cancer. In this study, we developed a novel surface-enhanced Raman spectroscopy (SERS)-based aptasensor to detect the prostate-specific antigen biomarkers, consisting of total prostate-specific antigen (PSA) and free prostate-specific antigen (f-PSA). A composite structure containing arrays of polystyrene colloidal sphere @Ag shell (PS@Ag) was fabricated as a SERS-active chip. A complementary DNA probe (SH-DNA) and PSA aptamer (Apt) were immobilised stepwise on the chip, followed by the binding of a Raman reporter methylene blue (MB) to the guanine base of the aptamer. PSA-Apt recognition causes the release of MB-Apt and a decrease in the SERS intensity of MB on the chip, which correlates with the PSA concentration. The proposed biosensor has high spectral reproducibility, selectivity, and sensitivity and successfully determines the PSA levels in serum samples collected from prostate cancer patients, demonstrating great potential for clinical diagnosis.

Study of the prostate-specific antigen–aptamer stability in the PSA–aptamer-single wall carbon nanotube assembly by docking and molecular dynamics simulation

ABSTRACT Prostate-specific antigen (PSA) is used to detect prostate cancer. RNA aptamers against this protein have been reported previously. In this work, the stability and interaction mechanism of aptamer with PSA and single-wall carbon nanotube (SWCNT) was studied by computational techniques. Molecular dynamics simulations were used to investigate the interaction of aptamer segments with the surfaces of SWCNT and prostate-specific antigen (PSA) protein in an aqueous solution by two steps. Firstly, the interaction of aptamer and SWCNT was studied by the molecular dynamic simulation method. Our results showed that an aptamer segment can attach to the surface of SWCNT with its helix axis perpendicularly. Our analysis shows that the π–π stacking interactions play an important role in the binding of the aptamer to carbon rings. Secondly, we analysed the interaction of PSA protein with aptamer in the aptamer–SWCNT assemble. We found that the 3D structure of aptamer in the presence of PSA protein and SWCNT was stable. Finally, this study provides evidence about the stability of aptamer as a sensor on the surface of SWCNT (as a fixed surface) for PSA protein as a ligand. This finding can help to improve new biosensors for detecting cancer protein markers.

Rapid and sensitive detection of prostate-specific antigen via label-free frequency shift Raman of sensing graphene

The sensitive and accurate detection of cancer biomarkers is critically important to early clinical diagnosis, disease monitoring, and successful cancer treatment. Here, we first demonstrate an aptamer-based frequency shift Raman approach via sensing of graphene. This biosensor allows the rapid, sensitive, and label-free detection of the acknowledged protein cancer biomarker, prostate-specific antigen (PSA). Monolayer graphene is employed as the Raman substrate, which is highly sensitive to its electronic structure and interface properties. The PSA aptamer can be adsorbed strongly on the surface of substrates through π-π stacking interactions. The vibrational frequency of the G peak of graphene shifted upon the specific binding between the PSA and its aptamer. The corresponding frequency shifts of the G peak are directly correlated with PSA concentrations. The limit of detection is as low as 0.01 ng/mL, with a wide linear range from 0.05 ng/mL to 25 ng/mL. The analytic samples can be detected directly without any extensive preparation and label process. The whole detection is completed in only 30 min. Furthermore, excellent recoveries are acquired to validate the feasibility of this assay in human serum samples. The proposed technology could provide a selective, versatile, and user-friendly strategy for the early detection of cancer biomarkers.

A cost-effective and rapid aptasensor with chemiluminescence detection for the early diagnosis of prostate cancer

A rapid aptasensor capable of early diagnosing prostate cancer was developed using a dual aptamer, which is composed of a prostate specific antigen (PSA) aptamer and hemin aptamer connected by a linker composed of 5 adenines. The dual aptamers immobilized on the surface of paramagnetic beads were able to competitively bind with PSA or hemin in human serum for 20 min at room temperature. After the incubation, the dual aptamers were separated from human serum using a magnetic separator. After the procedure, the mixture of Amplex Red and H2O2 in Tris–HCl (pH 8) was added in a microcentrifuge tube containing the dual aptamers. Resorufin was formed from the reaction of Amplex Red and H2O2 in the presence of horseradish peroxidase (HRP) mimicking DNAzyme formed from the reaction of hemin and the dual aptamer. The concentration of resorufin formed from the reaction was proportional to the concentration of HRP mimicking DNAzyme. Resorufin emitted bright red light with the addition of 1,1′-oxalyldiimidazole (ODI) chemiluminescence reagents such as ODI and H2O2. The brightness of the emission light was proportionally reduced with the increase of PSA in human serum due to the reduction of DNAzyme concentration with the binding of PSA and PSA aptamer linked to hemin aptamer. Using the calibration curve of the new aptasensor, trace levels of PSA in human serum was rapidly quantified for the early diagnosis of prostate cancer. The limit of detection (LOD = 3 s/slope) of the aptasensor was as low as 0.1 ng/ml. The accuracy, precision, and recovery of the aptasensor were studied with statistically acceptable methods.

TiO2 nanotubes loaded with CdS nanocrystals as enhanced emitters of electrochemiluminescence: application to an assay for prostate-specific antigen.

An enhanced cathodic electrochemiluminescence (ECL) assay for prostate-specific antigen (PSA) is developed based on the in situ activation of a semiconductor nanomaterial. An excellent ECL emitter (CdS/TiO2 nanotubes) was fabricated by the combination of TiO2 nanotubes (NTs) and thioglycolic acid-capped CdS nanocrystals (NCs). After the activation of the hydrogen peroxide-citric acid solution, the ECL signal was enhanced 265 times compared with that of the original TiO2 NT with H2O2 as co-reactant. For the ECL assay, activated CdS/TiO2 NTs were assembled with complementary DNA, PSA aptamer and probe DNA-functionalized SiO2@Pt nanoparticles (NPs) via DNA hybridization to form the detection platform. The SiO2@Pt NPs acted as ECL quencher of CdS/TiO2 NTs. In the presence of PSA, ECL increased after the release of pDNA-SiO2@Pt NPs because of the binding of PSA to the aptamer. An "off-on" ECL phenomenon appeared. The enhanced ECL signals were used for sensitive determination of PSA. The dynamic range was 0.001 to 50ngmL-1 with a detection limit of 0.4pgmL-1 (S/N = 3). This new approach conceivably paves the way for fabricating various other enhanced ECL emitter systems, with good application prospects in clinical practice. Graphical abstract The activated CdS/TiO2 nanotubes and SiO2@Pt nanoparticles were synthesized and used to develop an energy-transfer electrochemiluminescence analysis method with high sensitivity and anti-interference performance.

Construction of efficient “on-off-on” fluorescence aptasensor for ultrasensitive detection of prostate specific antigen via covalent energy transfer between g-C3N4 quantum dots and palladium triangular plates

Development of ultra-sensitive and high specific aptasensors is important for early diagnosis of prostate cancer. Herein, ultrasensitive detection of prostate specific antigen (PSA) aptasensor was realized based on the “on-off-on” model via fluorescence (FL) covalent energy transfer between g-C3N4 quantum dot (g-CNQDs) and palladium triangular plates (Pd TPs). Specifically, the Pd TPs were primarily linked with PSA aptamer (PA) as the reporter probe, followed by attaching them onto the g-CNQDs surfaces, causing the highly enlarged FL quenching rate (ca. 75%). After the introduction of PSA, the FL intensities recovered again because of the distinctively stronger affinity of PA to PSA than that of g-CNQDs. The bond of pyridine N with Pd was identified as efficient energy transfer pathway through the X-ray photoelectron spectroscopy (XPS) and FL measurements. The surface plasmon resonance (SPR) experiments certified the remarkably different affinity of PA towards g-CNQDs and PSA. The as-constructed FL aptasensor can accurately quantify PSA with wide linear range of 10 pg mL−1-50 ng mL−1 and ultra-low limit of detection (LOD, 4.2 pg mL−1), indicating the promising applications in clinical assay and biological detection.

Signal-switchable lab-on-paper photoelectrochemical aptasensing system integrated triple-helix molecular switch with charge separation and recombination regime of type-II CdTe@CdSe core-shell quantum dots

Herein, a new “on-off-on” signal switch system combined triple helix molecular switch with efficient charge separation and transfer between different sensitization units was designed for the ultrasensitive photoelectrochemical (PEC) determination of prostate-specific antigen (PSA). Concretely, the initial “signal-on” state was obtained via the cascaded sensitization structure consisting of type-II CdTe@CdSe core-shell quantum dots (QDs), CdS QDs, and ZnO nanotubes, which were assembled on Au nanoparticles modified paper fibers with the aid of signal transduction probe (STP). Thereinto, the type-II CdTe@CdSe QDs with hole-localizing core and electron-localizing shell could enable the ultrafast charge transfer and retard the charge recombination, magnifying the initial photocurrent response and preserving the high efficiency of signal-switchable PEC aptasensing system. Subsequently, the PSA aptamer (PSA-Apt) modified with gold nanoparticles (GNPs) was introduced by the hybridization of PSA-Apt with STP and the hairpin configuration of STP changed from closed to open state, forming a triple-helix structure. Hence, the CdTe@CdSe QDs labeled on the terminal of STP moved away from the electrode surface while the GNPs kept attached close to it. The proposed aptasensor turned to “signal-off” state because of the dual inhibition of vanished cosensitization effect and signal quenching effect of GNPs. Upon the target recognition, the triple-helix structure was perturbed with the formation of DNA-protein complex and the recovery of STP hairpin structure, resulting in the second “switch-on” state. Based on the target-induced photocurrent enhancement, the proposed PEC aptasensor was utilized for the determination of PSA with high sensitivity, persuasive selectivity, and excellent stability.

A label-free fluorescence method based on terminal deoxynucleotidyl transferase and thioflavin T for detecting prostate-specific antigen.

Prostate-specific antigen (PSA) is the only biomarker for the diagnosis of prostate cancer. So the PSA screening test is very important due to the high occurrence of prostate cancer in men. In this work, a label-free fluorescent method was developed based on terminal deoxynucleotidyl transferase (TdT) and G-quadruplex-thioflavin T complex for detecting PSA. In the absence of PSA, the PSA aptamer can be used as the primer for TdT extension reactions, resulting in the formation of G-quadruplexes and generation of strong fluorescent signals. After the addition of PSA, the PSA-aptamer complex prevented the TdT extension reaction due to steric hindrance, thus resulting in a poor fluorescent signal. The assay showed a wide linear range (0.1 to 80pg/mL) and a detection limit of 0.086pg/mL (S/N = 3). It also has good specificity for PSA determination and gives satisfactory results when applied to biological samples. Conceivably, its merits such as good selectivity and high sensitivity indicate that the proposed method has a promising application potential in the clinical diagnosis and treatment of prostate cancer. Graphical abstract.

Multicolor and photothermal dual-readout biosensor for visual detection of prostate specific antigen

Developing a facile and reliable approach for tumor marker detection is crucial in early diagnosis of cancer. Here, a multicolor and photothermal dual signal readout immunosensor was proposed based on a nanoparticle-mediated transformation strategy and used for sensitive detection of prostate specific antigen (PSA). To construct such a platform, Fe3O4 nanoparticles (NPs) -functionalized graphene oxide was modified with partially complementary DNA and served as the signal probes. In the absence of PSA, the signal probes were captured via hybridization reaction between DNA and PSA aptamer. Subsequently, Fe3O4 NPs anchored on the probes were transformed into a photothermal agent, Prussian blue NPs, which converted biological signal into heat via the near-infrared laser irradiation, and thus sensitive quantitative detection of PSA was realized by using a thermometer with a lower detection limit of 0.31 ng/mL. Meanwhile, Prussian blue NPs displayed multiply colors by mixing with potassium ferricyanide, and rapid qualitative detection by naked eyes was realized. The two sets of data mutually validate each other, which greatly improved the accuracy and reliability of PSA detection. More strikingly, graphene oxide as the enrichment carrier of Fe3O4 NPs significantly enhanced the accumulation of Prussian blue NPs in situ, and thus the signal amplification was effectively accomplished. Besides, the sensing strategy avoided the use of enzyme and simplified analysis process. Such a cost-effective and sensitive dual-readout visual protocol opens up new perspectives for personalized diagnosis and on-site detection.

The construction of electrochemical aptasensor based on coral-like poly-aniline and Au nano-particles for the sensitive detection of prostate specific antigen

Prostate specific antigen (PSA), as the significant biomarker of prostate cancer, has aroused widespread concern. However, constructing the PSA aptasensor with high sensitivity and low detection limit is still facing huge challenges. In this study, a sensitive electrochemical aptasensor for detecting PSA based on coral-like poly-aniline (PANI)/gold nano-particles (AuNPs) and the peptides was constructed. Coral-like PANI and AuNPs with the diameter of 50–80 nm, as the substrate materials, were directly deposited on glass carbon electrode (GCE) by electrodeposited method. Then, the peptides, as the antifouling materials, were anchored on the modified-electrode, which not only can form antifouling surface to reduce nonspecific adsorption, but also can be as a medium between the modified-electrode and PSA aptamer. Finally, the PSA aptamer was immobilized on the electrode. Based on the synergistic effect of excellent conductivity and special morphology of the substrate materials, and the antifouling ability of peptides, the aptasensor exhibited a high sensitivity of 462.7 μA(ng mL−1)−1 cm−2, wide linear range from 0.1 pg mL-1 to 100 ng mL-1, low limit of detection (LOD) of 0.085 pg mL-1 and good antifouling performance. The aptasensor also possessed outstanding selectivity, reproducibility and stability. Moreover, the aptasensor can directly detect PSA in 10% human serum solutions.

Emissions of terbium metal-organic frameworks modulated by dispersive/agglomerated gold nanoparticles for the construction of prostate-specific antigen biosensor.

Herein, a universal and multifunctional fluorescence sensor platform is designed by the interaction of aggregation/dispersion gold nanoparticles (AuNPs) with Tb-metal-organic frameworks (Tb-MOFs). It is found that the dispersed AuNPs rather than the aggregated ones can quench effectively the fluorescence of Tb-MOFs, and the quenching process presumably involves the mechanism of inner filter effect (IFE), dynamic quenching effect (DQE), and fluorescence resonance energy transfer (FRET). The different affinities of aptamer and aptamer-target complex toward AuNPs are employed to modulate the fluorescence signal change of Tb-MOFs. As the proof of concept, prostate-specific antigen (PSA), an efficient tumor indicator for prostate cancer, is selected as the target. At first, the PSA aptamer can protect AuNPs against salt-induced aggregation, leading to the fluorescence of Tb-MOFs quenching. Subsequently, upon PSA introduction, the rigid aptamer-PSA complex is formed and cannot stabilize AuNPs in high salt conditions, so the AuNPs aggregate significantly and the fluorescence of Tb-MOFs is restored. The linear range of PSA is achieved from 1 to 100ng/mL with a detection limit of 0.36ng/mL. Finally, this method has been validated to be sensitive and specific for PSA in human urine samples. Graphical abstract.

Hybridization induced fluorescence enhanced DNA-Ag nanocluster/aptamer probe for detection of prostate-specific antigen.

In this work, a label-free Ag nanocluster (AgNC)-based fluorescent probe is proposed to detect tumor marker, prostate-specific antigen (PSA). In the experiments, DNA sequences containing segments complemented to different parts of PSA aptamer were used to synthesize DNA-Ag nanoclusters (DNA-AgNC). Some of the obtained specific DNA-AgNC exhibited significant fluorescence increase after hybridization with PSA aptamer. Based on this, a simple DNA-AgNC/aptamer hybridization probe was fabricated for PSA detection using fluorescence quenching, because competitively specific binding between PSA and its aptamer inhibited the fluorescence enhancement effect of PSA aptamer on DNA-AgNC. The sequence of template DNA, pH and salt concentration of binding buffer, and the concentration of aptamer were optimized. Under optimum conditions, the concentration of PSA within the range of 2-150 ng mL-1 with the detection limit of 1.14 ng mL-1 was detected (3σ; n = 7). This approach was also successfully applied to determine PSA in spiked serum samples. As is well known, this was the first report to realize PSA detection using fluorescent AgNC-based probe. This work would provide reference for construction of AgNC-based probes for detecting other proteins.

Double Photosystems-Based 'Z-Scheme' Photoelectrochemical Sensing Mode for Ultrasensitive Detection of Disease Biomarker Accompanying Three-Dimensional DNA Walker.

A new double photosystems-based 'Z-scheme' photoelectrochemical (PEC) sensing platform is designed for ultrasensitive detection of prostate-specific antigen (PSA) by coupling with a three-dimensional (3D) DNA walker. Two photosystems consist of CdS quantum dots (photosystem I; PS I) and BiVO4 photoactive materials (photosystem II; PS II), whereas gold nanoparticles (AuNPs) photodeposited on high-active {010} facets of BiVO4 are used as the electron mediators to promote electron transfer from conduction band of PS II to valence band of PS I. 3D DNA walker-based amplification strategy is carried out between hairpin DNA1 conjugated onto the AuNP, hairpin DNA2 labeled with CdS quantum dot (QD-H2), and DNA walker complementary with the PSA aptamer modified to a magnetic bead (Apt-MB). Upon addition of target, DNA walker strand is displaced from DNA walker/Apt-MB to open hairpin DNA1 on AuNP@BiVO4. In the presence of QD-H2, DNA walker induces the hybridization of DNA1 with DNA2 on the gold nanoparticles step by step, thereby resulting in the assembly of CdS QDs on the AuNP@BiVO4 to form Z-scheme double photosystems with strong photocurrent. Under optimum conditions, the Z-scheme PEC sensing system exhibits good photocurrent responses toward target PSA within the working range of 0.01-50 ng mL-1 at a low detection limit of 1.5 pg mL-1. Good reproducibility and accuracy are acquired for analysis of target PSA and human serum specimens relative to the commercial PSA ELISA kit. Importantly, our strategy provides a new horizon for photoelectrochemical in vitro diagnostics.