Peroxidase-like Properties Research Articles

Nano-octahedral bimetallic Fe/Eu-MOF preparation and dual model sensing of serum alkaline phosphatase (ALP) based on its peroxidase-like property and fluorescence

Herein a nano-scale bimetallic Fe/Eu-MOF with a regular octahedral structure was synthesized for the first time. The synthesized Fe/Eu-MOF has both peroxidase-like activity and fluorescence properties. Fe/Eu-MOF can catalyze H2O2 to oxidize the chromogenic substrate TMB to produce blue oxTMB, which has ultraviolet absorption at 652 nm. Unexpectedly, the generated oxTMB can effectively quench the fluorescence of the catalyst Fe/Eu-MOF at 450 nm. The quenching mechanism is mainly the internal filtration effect (IFE), accompanied by static quenching (SQE), Förster resonance energy transfer (FRET) and photoelectron transfer (PET). Fe/Eu-MOF has a high affinity for sodium pyrophosphate (PPi). PPi can be adsorbed to the surface of Fe/Eu-MOF, destroying the structure of Fe/Eu-MOF and inhibiting its catalytic activity, resulting in a decrease in UV absorbance and the decline of fluorescence quenching. In contrast, phosphoric acid (Pi) has almost no effect on the reaction system. Alkaline phosphatase (ALP) can catalyze the hydrolysis of PPi to Pi, thereby reducing the inhibitory effect of PPi. Based on this, we successfully constructed a dual-mode ALP sensor with high selectivity. The linear ranges based on the 652 nm absorption or the fluorescence detection are from 1 to 200 U/L, and the detection limits are 0.6 for the absorption method and 0.9 U/L for the fluorescence method, respectively.

Sophisticated plasmon-enhanced photo-nanozyme for anti-angiogenic and tumor-microenvironment-responsive combinatorial photodynamic and photothermal cancer therapy

In the exploitation of nanozymes possessing intrinsic enzyme-like activities for cancer therapy, minor focus has been devoted to plasmonic nanostructures with localized surface plasmon resonance (LSPR)-driven properties. Here, we report the application of unique peroxidase-mimicking plasmonic photo-nanozymes coupling tumor-microenvironment-responsive reactive oxygen species generation with photothermal effect for effective combinatorial therapy. The well-defined anisotropic photo-nanozyme is synthesized by selectively depositing Pd nanoparticles on the tips of gold nanobypyramids. Intrinsic peroxidase-like properties with 1.5-fold-activity enhancement under photoexcitation are ascribed to a Pd-induced hot electrons/holes separation with efficient H2O2 decomposition. The LSPR-induced photocatalytic/photothermal combinatorial effects are remarkably enhanced upon H2O2 addition, critically suppressing the cell survival rate under near-infrared light. An effective decomposition of cell-signaling H2O2 additionally reveals prominent expression hindrance of vascular endothelial growth factor and hypoxia-inducible factor 1α. Our seminal findings uncover an interrelation between LSPR-induced phenomena and biomimetic fingerprints, valuable to overcome the shortcomings of conventional photodynamic therapy.

Hierarchical micro-nanostructures from polyoxometalates and polydopamine: Characterization, electrochemical and intrinsic peroxidase-like properties

Hybrid composite materials with hierarchical structures have attracted continuing attention for their enhanced features and various applications. The new hybrid composites XW9@PDA (XW9 means Na8H[α-PW9O34], Na10[α-SiW9O34] and Na10[α-GeW9O34]; PDA means polydopamine) have been synthesized for potential sensing applications. The effect of polyoxometalates on the formation of the hybrid composites was investigated. Scanning electron microscopy (SEM) supported the obtained XW9@PDA were hierarchical microsphere. By monitoring the reaction of classical peroxidase substrate o-phenylenediamine (OPD) to form yellow 2, 3-diaminophenazine (DAB) in the presence of H2O2, the peroxidase-like activity of XW9@PDA was calculated. The results showed that XW9@PDA still keep the peroxidase-like activity and have 505.98%, 208.58% and 582.78% enhancement in intrinsic catalytic activity for PW9@PDA, SiW9@PDA and GeW9@PDA. Among the three composites, PW9@PDA showed the highest peroxidase-like activity. Therefore, PW9@PDA was applicable to the colorimetric and electrochemical sensing for H2O2.

Glypican-3 electrochemical aptasensor based on CuO-rGO nanocomposite

Herein, a novel electrochemical immunosensor was fabricated based on copper oxide-reduction graphene oxide (CuO-rGO) nanocomposites for Glypican-3 (GPC3) detection. Firstly, CuO-rGO nanocomposite as nanoenzyme with good peroxidase-like properties is synthesized by One-step hydrothermal. Then CuO-rGO nanoenzyme captures GPC3 aptamer to form a signal probe, finally GPC3 antibody is used as the recognition probe to form an "antibody-antigen-aptamer" sandwich structure. Due to the peroxidase-like properties of the probe, this sandwich structure can effectively catalyze the reaction between hydrogen peroxide (H2O2) and silver nitrate (AgNO3) and AgNPs is deposited on the surface of the electrode to amplify the final measured current. The Differential Pulse Voltammetry (DPV) method was employed to record the response signal of the immunosensor. The current response presented a linear relationship to the GPC3 concentration in the range of 1 to 40 μg/mL with the detection limit of 0.45 μg/mL.

A visible and colorimetric nanobiosensor based on DNA-CuO nanoparticle for detection of single nucleotide polymorphism involved in sickle cell anemia disease

Sickle cell anemia (SCA) is a blood monogenic disease, which is caused by a single nucleotide mutation in the β-globin gene. In this work, to diagnose prenatal of the disease, a colorimetric nanobiosensor established using catalytic and peroxidase-like properties of copper oxide nanoparticle (CuO) and its interaction with DNA. It used to detect SNP in cell-free fetal DNA (cffDNA), as a promising diagnostic biomarker. The nanobiosensor detected the target sequence of the non-target, exactly even with one nucleotide polymorphism. The optimized method was capable of detecting the target sequence over its concentrations ranged from 2 nM to 12 nM (R2 = 0.96) with a detection limit of 0.64 nM within a time of 10 min. To conclude, the SNP detection based on this method shows a cheap, fast, visible, and versatile opportunity in sensitive and selective prenatal diagnosis of hereditary diseases with good reproducibility and stability.

Biocompatible Platinum Nanoclusters Prepared Using Bitter Gourd Polysaccharide for Colorimetric Detection of Ascorbic Acid.

Ascorbic acid is an organic compound with antioxidant properties that can protect the human body from the threat of free radicals. Therefore, it is important to detect the existence and measure the concentration of ascorbic acid to regulate its content in the human body. In this work, we prepared bitter gourd polysaccharide (BGP)-stabilized platinum nanoclusters (Pt-BGP NCs) by reacting BGP with K2PtCl4. Pt-BGP NCs and catalyzed the decomposition of H2O2 to generate •OH radicals, which could oxidize TMB to form oxidized TMB (oxTMB), indicating their peroxidase-like properties. The kinetics followed the Michaelis–Menten equation. Furthermore, the colorimetric detection of ascorbic acid using Pt-BGP NCs showed high selectivity and a low detection limit of 0.191 μM. The accuracy of real sample detection using Pt-BGP NCs was as high as 98.9%. More importantly, Pt-BGP NCs had high cell biocompatibility and extremely low hemolysis rate due to the component of BGP. In summary, the prepared Pt-BGP NCs with reductive activity and good biocompatibility have good application prospects in colorimetric detection of ascorbic acid.

Facile and selective colorimetric assay of choline based on AuNPs-WS2QDs as a peroxidase mimic

In the current work, we have fabricated a paper test strip for visual assay of choline by using choline oxidase enzyme with H2O2 based on the oxidation product of peroxidase substrate 3,3′,5,5′-tetramethylbenzidine (TMB) catalyzed by gold nanoparticles decorated WS2 quantum dots (AuNPs-WS2QDs) to appear blue color product. Under a large range of temperature, pH and H2O2 concentration, AuNps-WS2QDs exhibit prominent peroxidase-like property. In the presence of oxygen, choline oxidized by choline oxidase which produces H2O2 is colorimetrically measured, at 652 nm. The blue color appears due to biochemical reactions between TMB and AuNps-WS2QDs were used as an indication for estimation of H2O2. The proposed scheme is applied for choline detection in concentration range of 1–150 μM in PBS with the 0.086 µM limit of detection. Color variation can be observed visually and this assay is selective, simple and quick. Further portable test kit was fabricated for choline detection in real samples i.e. serum and milk which shows an excellent response towards choline detection. The filter paper test strip changed color intensity based on the concentration of choline.

Nanomaterial-enhanced 3D-printed sensor platform for simultaneous detection of atrazine and acetochlor

The widespread use of herbicides in agriculture and gardening causes environmental and safety issues such as water pollution. Thus, efficient and convenient analysis of the levels of herbicide residues is of significant importance. Here, we employed 3D-printing to design a multiplex immunosensor for simultaneous detection of two widely used herbicides, atrazine and acetochlor. Multiplexing was achieved through customization of a lateral flow immunoassay, and then integrated with an electrochemical analyzer for ultrasensitive detection. Quantification of herbicide residues was realized through the detection of a novel nanomaterial label, the mesoporous core-shell palladium@platium nanoparticle (Pd@Pt NP), for its outstanding peroxidase-like property. During the electrochemical analysis, the catalytic activity of Pd@Pt NPs on the redox reaction between thionin acetate and hydrogen peroxide provided an electrochemically driven signal that accurately indicated the level of herbicide residues. Using this Nanomaterial-enhanced multiplex electrochemical immunosensing (NEMEIS) system, simultaneous detection of atrazine and acetochlor was realized with a limit of detection of 0.24 ppb and 3.2 ppb, respectively. To further evaluate the feasibility, the optimized NEMEIS was employed for detection in atrazine and acetochlor residue-containing spiked samples, and an overall recovery with 90.8% – 117% range was obtained. The NEMEIS constructed with the aid of 3D-printing provides a rapid, precise, economical, and portable detection device for herbicides, and its success suggests potential broad applications in chemical analysis, biosensors and point-of-care monitoring.

Green synthesis of stable platinum nanoclusters with enhanced peroxidase-like activity for sensitive detection of glucose and glutathione

The sensitive and facile detection of the concentration of glucose and glutathione (GSH) is important for our health. In this work, lily polysaccharide (LP) stabilized platinum nanoclusters (Ptn-LP NCs) were prepared by a facile synthetic strategy where LP was used as a reducing agent and stabilizer. The successful preparation of 1–2 nm platinum nanoclusters (Pt NCs) inside of Ptn-LP NCs was confirmed using UV–Vis spectra, X-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectroscopy (FTIR) and Transmission electron microscopy (TEM), respectively. Furthermore, Ptn-LP NCs accelerated the catalytic oxidation of colorless 3,3′,5,5′-tetramethylbenzidine (TMB) to blue oxidized TMB (oxTMB) in the presence of hydrogen peroxide (H2O2), indicating their intrinsic peroxidase-like activity. The Vm values of TMB and H2O2 were 50.3 × 10−8 Ms−1 and 10.2 × 10−8 Ms−1, respectively, which were higher than those of other mimetic enzymes. The peroxidase-like property of Ptn-LP NCs was used to sensitively detect the concentration of glucose and glutathione (GSH). The detection limits of glucose and GSH were as low as 1.89 and 0.37 μM, the linear ranges of glucose and GSH were 2.5–1000 and 4–140 μM, respectively. The colorimetric method was successfully applied to detect GSH concentration in human serum. This method has great potential application in bio-related detection in the future.

Magnetic rod-based metal-organic framework metal composite as multifunctional nanostirrer with adsorptive, peroxidase-like and catalytic properties

Although magnetic stirring is frequently used to enhance the kinetics for adsorption, chemical and biochemical reactions, the introduction of stirrers inevitably leads to the adsorption of analytes and thus interferes with the efficiency of the chemical process or reaction. In this work, magnetic Fe3O4 nanorods with tunable length-to-diameter ratio were synthesized via a hydrothermal method and used as templates for the in-situ depositing of MIL-100(Fe) and gold nanoparticles. Such nanorod-based material can not only function as an adsorbent, nanozyme, and a heterogeneous catalyst for corresponding applications but also serve as a magnetic nanostirrer to enhance kinetics. As a proof-of-concept, the capture of bacteria pathogen, mimic-peroxidase-based colorimetric detection of hydrogen peroxide, and the catalytic reduction of selected organic pollutants were conducted using the as-synthesized Fe3O4@MIL-100(Fe)-Au nanostirrer with and without magnetic field. The results show that the rates of bacteria capture, mimetic enzyme reaction and catalysis were tremendously expedited. We believe this magnetic field-assisted approach holds great promise for future applications, because, not only does it eliminate the use of external magnetic stirrers and thereby decrease the risk of foreign pollution but also, is adaptable for nanoscale reaction systems where conventional stirring is not applicable due to size limitations.

Two-Dimensional MnO2 Nanozyme-Mediated Homogeneous Electrochemical Detection of Organophosphate Pesticides without the Interference of H2O2 and Color.

Traditional peroxidase-like nanozyme-based sensors suffer from self-decomposition and high toxicity of H2O2, as well as the interference of color from nanozymes themselves and testing samples. In this work, we adopt nanozymes (two-dimension (2D) MnO2 sheets, manganese dioxide nanosheets (MnNS)) with oxidase-like and peroxidase-like properties as advanced catalysts to develop a novel homogeneous electrochemical sensor for organophosphate pesticides (OPs) using dissolved O2 as a coreactant without the interference of H2O2 and color. Owing to the large surface area and unique catalytic activity of MnNS, a large amount of tetramethylbenzidine (TMB) is catalyzed oxidation, leading to a significantly declined differential pulse voltammetry (DPV) current. Obviously, MnNS display an excellent response to thiocholine, deriving from the catalyzing hydrolysis of acetylthiocholine (ATCh) by acetylcholinesterase (AChE), which switches a homogeneous electrochemical OP detection process based on the depressing AChE activity with a limit of detection (LOD) of 0.025 ng mL-1. The as-proposed strategy on using nanozymes with oxidase-like and peroxidase-like properties to develop a homogeneous electrochemical sensor will provide a new pathway for improving the performance of nanozyme-based sensors, and the established MnNS-based homogeneous electrochemical sensor will find more applications for OP residue determination in food samples.

Investigation of efficient synergistic and protective effects of chitosan on copper nanoclusters: Construction of highly active and stable nanozyme for colorimetric and fluorometric dual-signal biosensing

Copper nanoclusters (CuNCs) have advantages of low cost, excellent biocompatibility, and good fluorescence properties, but they are seldom used as nanozymes for biosensing owing to their low catalytic activity and poor stability. Herein, a natural polymer, chitosan (CS), was explored to effectively promote the peroxidase-like property of glutathione-protected copper nanoclusters (GSH-CuNCs). Due to the synergistic catalytic enhancement and protection effect provided by CS, both peroxidase-like activity and stability of CS@GSH-CuNCs were significantly improved compared with those of GSH-CuNCs. Moreover, upon the addition of H2O2, CS@GSH-CuNCs can catalyze the oxidation of colorless 3, 3′, 5, 5′-tetramethylbenzidine (TMB) to produce oxidized TMB (oxTMB) with blue color, meanwhile, H2O2 can efficiently cause the fluorescence quenching of CS@GSH-CuNCs. Based on that, a dual-signal approach was designed for the detection of H2O2, choline, and acetylcholinesterase (AChE) activity. For the colorimetric assay, this method exhibits detection limit (DL) of 6.7 μM for H2O2, 6.5 μM for choline, and 3.3 mU/mL for AChE activity. For the fluorometric assay, this approach presents DL of 0.6 μM for H2O2, 1.2 μM for choline, and 1.4 mU/mL for AChE activity. Furthermore, this dual-signal assay was applied to milk and human serum, indicating its great prospect for real sample analysis.

Green synthesis of platinum nanoclusters using lentinan for sensitively colorimetric detection of glucose

The sensitive colorimetric detection of glucose using nanomaterials has been attracting considerable attention. To improve the detection sensitivity, highly stable lentinan stabilized platinum nanoclusters (Pt-LNT NCs) were prepared, in which lentinan was employed as a mild reductant and stabilizer. The size of platinum nanoclusters (Pt NCs) was only 1.20 ± 0.29 nm. Pt-LNT NCs catalyzed the oxidation of substrate 3,3′,5,5′-tetramethylbenzidine (TMB) in the presence of hydrogen peroxide (H2O2) to produce a blue oxidation product with absorption peak at 652 nm, indicating their peroxidase-like properties. Their enzymatic kinetics followed typical Michaelis-Menten theory. In addition, fluorescence experiments confirmed their ability to efficiently catalyze the decomposition of H2O2 to generate •OH, which resulted in the peroxidase-like mechanism of Pt-LNT NCs. Moreover, a colorimetric method for highly selective and sensitive detection of glucose was established by using Pt-LNT NCs and glucose oxidase. The linear range of glucose detection was 5–1000 μM and the detection limit was 1.79 μM. Finally, this method was further used for detection of glucose in human serum and human urine. The established colorimetric method may promote the development of biological detection and environmental chemistry in the future.

(Z,Z)-Selanediylbis(2-propenamides): Novel Class of Organoselenium Compounds with High Glutathione Peroxidase-Like Activity. Regio- and Stereoselective Reaction of Sodium Selenide with 3-Trimethylsilyl-2-propynamides.

The efficient regio- and stereoselective synthesis of (Z,Z)-3,3′-selanediylbis(2-propenamides) in 76–93% yields was developed based on the reaction of sodium selenide with 3-trimethylsilyl-2-propynamides. (Z,Z)-3,3′-Selanediylbis(2-propenamides) are a novel class of organoselenium compounds. To date, not a single representative of 3,3′-selanediylbis(2-propenamides) has been described in the literature. Studying glutathione peroxidase-like properties by a model reaction showed that the activity of the obtained products significantly varies depending on the organic moieties in the amide group. Divinyl selenide, which contains two lipophilic cyclohexyl substituents in the amide group, exhibits very high glutathione peroxidase-like activity and this compound is considerably superior to other products in this respect.

Through quorum sensing, Pseudomonas aeruginosa resists noble metal-based nanomaterials toxicity.

Noble metal-based nanomaterials (NMNs), such as platinum nanoparticles (Pt@NPs) and palladium nanoparticles (Pd@NPs), are increasingly being used as antibacterial agents. However, little information is available on bacterial resistance to NMNs. In this study, owing to their oxidase-like and peroxidase-like properties, both Pt@NPs and Pd@NPs induce reactive oxygen species (ROS) and manifest antibacterial activities: 6.25μg/mL of either Pt@NPs or Pd@NPs killed >50% of Staphylococcus aureus strain ATCC29213. However, Pseudomonas aeruginosa strain PAO1 completely resisted 12.5μg/mL of Pt@NPs and 6.25μg/mL of Pd@NPs. Compared to the non-NMN groups, these NMNs promoted 2-3-fold upregulation of the quorum sensing (QS) gene lasR in strain PAO1. In fact, the lasR gene upregulation induced a 1.5-fold reduction in ROS production and increased biofilm formation by 11% (Pt@NPs) and 27% (Pd@NPs) in strain PAO1. The ΔlasR mutants (lasR gene knock out in strain PAO1), became sensitive to NMNs. The survival rates of ΔlasR mutants at 12.5μg/mL Pt@NPs and Pd@NPs treatments were only 77% and 58%, respectively. This is the first report indicating that bacteria can resist NMNs through QS. Based on these results, evaluation of the ecological risks of using NMNs as antibacterial agents is necessary.

Fluorometric Detection of Thiamine Based on Hemoglobin-Cu3(PO4)2 Nanoflowers (NFs) with Peroxidase Mimetic Activity.

Component analysis plays an important role in food production, pharmaceutics and agriculture. Nanozymes have attracted wide attention in analytical applications for their enzyme-like properties. In this work, a fluorometric method is described for the determination of thiamine (TH) (vitamin B1) based on hemoglobin–Cu3(PO4)2 nanoflowers (Hb–Cu3(PO4)2 NFs) with peroxidase-like properties. The Hb–Cu3(PO4)2 NFs catalyzed the decomposition of H2O2 into ·OH radicals in an alkaline solution that could efficiently react with nonfluorescent thiamine to fluoresce thiochrome. The fluorescence of thiochrome was further enhanced with a nonionic surfactant, Tween 80. Under optimal reaction conditions, the linear range for thiamine was from 5 × 10−8 to 5 × 10−5 mol/L. The correlation coefficient for the calibration curve and the limit of detection (LOD) were 0.9972 and 4.8 × 10−8 mol/L, respectively. The other vitamins did not bring about any obvious changes in fluorescence. The developed method based on hybrid nanoflowers is specific, pragmatically simple and sensitive, and has potential for application in thiamine detection.

A novel dual signal and label-free electrochemical aptasensor for mucin 1 based on hemin/graphene@PdPtNPs

A dual signal and label-free electrochemical aptasensor for mucin 1 was constructed based on hemin/graphene@PdPtNPs nanocomposite (H-Gr@PdPtNPs). Hemin attached on the graphene surface not only improves the solubility of graphene and acts as an in-situ electrochemical probe but also exhibits excellent peroxidase-like properties to electrocatalyze the reduction of H2O2. PdPtNPs also show outstanding catalytic capacity to the reduction of H2O2 and provide numerous binding sites for loading dDNA (mucin 1 aptamer and cDNA) to form the sensing interface. In the presence of mucin 1, due to the specific affinity between aptamer and mucin 1, double helix would be induced dissociation and the aptamer would be pulled off from the electrode. As a result, the electrochemical signals of hemin and H2O2 were recovered. Based on these properties, the label-free and sensitive dual signal electrochemical biosensor for mucin 1 detection has been developed. The one is differential pulse voltammetry (DPV) signal of hemin and the other is chronoamperometry signal arisen from the catalytic reduction of H2O2. The linear ranges for mucin 1 were 8.0 pg mL−1 to 80 ng mL−1 and 0.8 pg mL−1 to 80 ng mL−1 with the limit of detection 2.5 pg mL−1 and 0.25 pg mL−1 by DPV and chronoamperometry, respectively. The recovery of mucin 1 in human blood serum samples was from 95.0% to 104.2%. The detection platform does not need signal labeling which greatly reduced the sophisticated and expensive procedures. The aptasensor provide a promising strategy for the determination of mucin 1 in clinical diagnostics.

A nanocomposite hydrogel with catalytic properties for trace-element detection in real-world samples

A nanocomposite material characterized by peroxidase-like properties was developed through the dispersion of platinum nanoparticles (PtNPs) inside a hydrogelic matrix. The integration of the PtNP catalysts within the matrix resulted in their stabilization, preventing aggregation and precipitation in media of environmental interest, characterized by high ionic strength and by the presence of organic solutes. A thorough optimization of the matrix design was aimed at granting optimal diffusion of the reagents, in order to maintain the efficiency of the catalytic action. Such combined features allowed the implementation and prototyping of a colorimetric method for the detection of mercury in environmental water samples. The assay was based on a chromogenic reaction catalyzed by the peroxidase-like activity of PtNPs and its specific inhibition caused by trace amounts of mercury.

Immunoassay using dendritic Au-Pt nanoparticles as signal labels for detection of the biomarker of Burkholderia pseudomallei

In this work, the peroxidase-like properties of dendritic Au-Pt nanoparticles (30~60 nm) were investigated and these nanoparticles were applied in the immunoassay as signal labels for the detection of capsular polysaccharides (CPS, ~ 300 kDa, expressed on the cell envelopes of Burkholderia pseudomallei causing melioidosis). The immunoassay adopts monoclonal anti-CPS antibodies. The assay limit of detection (LOD), which is calculated through the 3σ method, is at 0.36 ng/mL for CPS spiked in PBS with 5% milk, and at 0.88 ng/mL for CPS spiked in human serum. The results from this work show that the dendritic Au-Pt nanoparticles can produce similar assay performance in detecting CPS as HRP did. Considering these particles can be easily prepared and stable in environments (thus avoiding cold-chain based storage/transportation), they have potential to be applicable for both in-house and in-field immunoassay development for the pathogen B. pseudomallei detection.

Platinum nanoflowers with peroxidase-like property in a dual immunoassay for dehydroepiandrosterone.

Platinum nanoflowers (PtNFs) were utilized in a competitive enzyme-linked immunosorbent assay (ELISA) and in a lateral flow immunoassay (LFIA) for superior peroxidase-like activity and intense brown color, respectively. PtNFs were linked to the polyclonal antibody (pAb) to form the pAb-PtNFs probes for the dual immunoassay. Based on optimized pAb-PtNF probes, both enzyme-linked immunosorbent assay (PtNFs-ELISA) and lateral flow immunoassay (PtNFs-LFIA) perform very well. The absorbance at 450nm decreases linearly in the DHEA concentration range 2.1 to 118.1ngmL-1, and the limit of detection is 1.3ngmL-1 and theIC50 value is 15.7ngmL-1 of PtNFs-ELISA. The visual cut-off value of PtNFs-LFIA is 10.0ngmL-1. The average recoveries from spiked samples range from 95.0 to 108.9% with a coefficient ofvariation below 12.2%. Excellent recoveries and correlation between the two methods were observed. Furthermore, the designed immunosensors exhibited good selectivity, confirming a broad development prospect in DHEA monitoring. Graphical Abstract.