Label-free Electrochemical Biosensor Research Articles

A novel label-free impedance biosensor for KRAS G12C mutations detection based on PET-RAFT and ROP synergistic signal amplification

The KRAS G12C mutations, as crucial biomarkers, are closely associated with non-small cell lung cancer. Here, a novel label-free electrochemical biosensor with synergistic signal amplification of photocell energy transfer-reversible addition fragmentation chain transfer (PET-RAFT) and ring-opening polymerization (ROP) was developed for the first time for sensitive detection of KRAS G12C mutations. Specifically, hairpin DNA (hDNA), which act as biomolecular probe, was self-assembled on Au electrode surface by Au-S bond. 4-cyano-4-[(dodecylsulfanylthiocarbonyl) sulfanyl] pentanoic acid (CDTPA), the chain transfer agent of PET-RAFT reaction, was then attached to hDNA via amide bond. After that, the target DNA (tDNA) was captured on the electrode surface by complementary base pairing with hDNA. Subsequently, large numbers of electro-active monomers N-acryloxysuccinimide (NAS) were successfully grafted to the electrode surface via PET-RAFT reaction, which provided plenty of junction sites for doxorubicin-polycaprolactone (Dox-PCL) synthesized by ROP. Finally, the Dox-PCL was connected to the electrode surface by ester bond, significantly amplifying the electrochemical signal. Under optimized conditions, the biosensor has a wide linear detection range of 0.1 pM to 1 μM, with a detection limit of 86.9 fM. Attribute to its high sensitivity, specificity, reproducibility and stability, this biosensor possesses considerable potential in early diagnosis of disease and biomedical research.

Bioreceptors’ immobilization by hydrogen bonding interactions and differential pulse voltammetry for completely label-free electrochemical biosensors

Label-free electrochemical biosensors show great potential for the development of point-of-care devices (POCDs) for environmental and clinical applications. These sensors operate with shorter analysis times and are more economic than the labelled ones. Here, four completely label-free biosensors without electron transfer mediators were developed for hepatitis B virus (HBV) detection. The approach consisted in (i) the modification of gold surfaces with cysteamine (CT) or cysteine (CS) linkers, (ii) the subsequent antibody (Ab) immobilization, either directly by hydrogen bonding (HB) interactions or by covalent bonds (CB) using additional reagents, and (iii) measuring the biosensor response by electrochemical impedance spectroscopy (EIS) and differential pulse voltammetry (DPV). The electrode surfaces at each stage of the modification process were characterised by X-ray photo-electron spectroscopy (XPS) and atomic force microscopy (AFM). The combination of Ab immobilization by HB with the DPV analysis displayed improved repeatability, lower interference to serum matrix and similar limits of detection and quantification than the traditional biosensors that immobilize the Ab via CB and use EIS as readout technique. The Ab immobilization by HB is shown as a simple, efficient and low-cost alternative to CB ones, while DPV was faster and showed better performance than EIS. The CT-HB biosensor displayed the lowest limits of detection and quantification of 0.14 and 0.46 ng/mL, respectively, a 0.46–12.5 ng/mL linear analytical range, and 100% of recovery for 1/10 human serum media during HBV surface antigen detection by DPV. Even, it preserved the initial sensing capability after 7 days of its fabrication.Graphical abstract

Impedimetric Gram-Positive Bacteria Biosensor Using Vancomycin-Coated Silica Nanoparticles with a Gold Nanocluster-Deposited Electrode.

We introduce a swift, label-free electrochemical biosensor designed for the precise on-site detection of Gram-positive bacteria via electrochemical impedance spectroscopy. The biosensor was prepared by electroplating the electrode surface with gold nanoclusters (AuNCs) on the gold-interdigitated wave-shaped electrode with a printed circuit board (Au-PCB) electrode, which plays a role in cost-effective and promising lab-on-a-chip microsystems and integrated biosensing systems. This was followed by the application of silica nanoparticle-modified vancomycin (SiNPs-VAN) that binds to Gram-positive bacteria and facilitates their detection on the AuNC-coated surface. The biosensor demonstrated remarkable sensitivity and specificity. It could detect as few as 102 colony-forming units (CFU)/mL of Staphylococcus aureus, 101 CFU/mL of Bacillus cereus, and 102 CFU/mL of Micrococcus luteus within 20 min. Additionally, SiNPs-VAN is also known for its high stability, low cost, and ease of preparation. It is effective in identifying Gram-positive bacteria in water samples across a concentration range of 102-105 CFU/mL and shows selective identification of Gram-positive bacteria with minimal interference from Gram-negative bacteria like Escherichia coli. The ability of the biosensor to quantify Gram-positive bacteria aligns well with the results obtained from the quantitative real-time polymerase chain reaction (qRT-PCR). These findings highlight the potential of electrochemical biosensors for the detection of pathogens and other biological entities, marking a significant advancement in this field.

Diagnosis of viral infectious diseases through sensitive detection of human serum antibodies using a modified label-free electrochemical biosensor.

Rapid virus identification is crucial for preventing outbreaks. The COVID-19 pandemic has highlighted the critical nature of rapid virus detection. Here, we designed a label-free electrochemical biosensor modified with gold nanoparticles (AuNPs) to detect IgG antibodies from human serum, enabling rapid point-of-care diagnostics. AuNPs were synthesized and characterized. A multivariate optimization was carried out to determine the optimal condition for functionalizing AuNPs with anti-IgG. Subsequently, using a glassy carbon electrode (GCE), a modified AuNPs/GCE electrochemical biosensor was developed for IgG detection. The results indicated thatAuNPs displayed a spherical morphology with a size distribution of 19.54nm. Additionally, the zeta potential was recorded at -7.84mV. Central composite design (CCD) analysis determined the optimal conditions for functionalizing AuNPs to be an anti-IgG concentration of 320µgmL-1, atemperature of 25°C, and pH of 7.4. The characterization study confirmed the successful synthesis and functionalization of AuNPs. Through electrochemical impedance spectroscopy measurement, the biosensor demonstrated a limit of detection (LOD) of 0.2ngmL-1 and limit of quantification (LOQ) of 0.8ngmL-1. Furthermore, tests in real samples showed the interaction between IgG antibodies in serum samples and AuNPs/GCE, confirming the biosensor's ability to detect and quantify IgG in clinical samples.

Novel facile fabrication of a label-free femto-molar serotonin electrochemical biosensor based on functional MoS2 nano flakes

Our bodies contain a neurotransmitter called serotonin, which regulates several activities connected to our temporary state of mind or temper. Serious disorders linked to mood disorders, anxiety, and depression are diagnosed due to serotonin disturbance. If diagnosed early, serotonin fluctuations could be controlled through proper medication. We present the electrochemical detection of serotonin molecules at the femtomolar level using carboxylate molybdenum disulfide (MoS2-COOH) functionalized on the gold electrode surface. X-ray diffraction spectroscopy explores the synthesized MoS2-COOH conjugates structural properties, and their functional behavior is confirmed by FT-IR study. The square wave voltammetry confirmed that the carboxylated MoS2 nanocomposite-modified electrode offered excellent electrocatalytic activity towards serotonin oxidation. The sensor excels in sensing serotonin over a wide dynamic range from 1 µM to 1 fM, with a limit of detection of 12.038 fM, excellent reproducibility, and high selectivity towards serotonin in interfering medium containing dopamine. This sensitive, selective, and cost-effective electrochemical sensor shows significant potential in direct serotonin analysis for clinical applications.

A single-use electrochemical biosensor system for ultrasensitive detection of Aflatoxin B1 in rice, corn, milk, peanut, chili pepper samples

Aflatoxin B1, a common food contaminant in peanuts and corn and a genotoxic carcinogen in humans poses a significant risk for hepatocellular carcinoma, making its detection crucial; this study aims to develop a label-free electrochemical biosensor using a disposable indium tin oxide polyethylene terephthalate (ITO-PET) electrode modified with 3-Aminopropyltriethoxysilane for detecting Aflatoxin B1 in real food samples. Initially, optimization steps for the proposed biosensor were conducted using electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) techniques. Characterization steps such as storage capacity, regeneration, and single frequency impedance (SFI) were completed for the proposed disposable biosensor after the optimization steps. The electrochemical biosensor, based on AFB1, exhibited excellent repeatability and reproducibility. It had a broad dynamic detection range from 0.1 fg/mL to 500 fg/mL, with low limits of detection (LOD) and quantitation (LOQ) at 0.19 fg/mL and 0.65 fg/mL, respectively. Finally, the proposed AFB1-based biosensor system was applied to real food samples (rice, chili pepper, milk, corn, and peanuts) for testing and validation.

A bulged-type enzyme-free recognition strategy designed for single nucleotide polymorphisms integrating with label-free electrochemical biosensor

Compared to conventional nucleic acid detection methods, label-free single nucleotide polymorphism (SNP) detection presents challenging due to the necessity of discerning single base mismatches, especially in the field of enzyme-free detection. In this study, we introduce a novel bulged-type DNA duplex probe designed to significantly amplify single-base differences. This probe is integrated with programmable DNA-based nanostructures to develop a sensitive, label-free biosensor for nonenzymatic SNP detection. The duplex probe with one bulge could selectively identify wild-typed DNA (WT) and mutant-type DNA (MT) based on a competitive strand displacement reaction mechanism. The hyperbranched HCR (HHCR) by incorporating of hairpin DNA into the DNA tetrahedron and surface-tethering on the portable screen printing electrode (SPCE) significantly favor the formation of negatively charged DNA nanostructure. We harnessed strong repulsion of DNA nanostructure towards the electroactive [Fe(CN)₆]³⁻/⁴⁻ in combination with electrochemical technique to create a label-free biosensor. This simple, enzyme-free and label-free biosensor could detect MT with a detection limit of 56 aM, even in multiple sequence backgrounds. The study served as the proof-of-concept for the integration of enzyme-free competitive mechanism and label-free strategy, which can be extended as a powerful tool to various fields.

Ultrasensitive detection of CEA in human serum using label-free electrochemical biosensor with magnetic self-assembly based on α-Fe2O3/Fe3O4 nanorods

Early-stage colorectal cancer (CRC) is often asymptomatic, leading to late diagnoses and impacting treatment outcomes. Timely detection and diagnosis are crucial, with serum carcinoembryonic antigen (CEA) levels playing a key role in CRC diagnosis and prognosis. To enhance precision, a label-free electrochemical nano-biosensor utilizing magnetic self-assembly (MSA) technology was developed for sensitive CEA detection. The biosensor's core was the α-Fe2O3/Fe3O4 nanorods prepared by the hydrolysis-calcination reduction process. Its unique magnetism was the cornerstone of MSA technology. The biosensor exhibited excellent mechanical strength, ensuring stability during storage at 4 °C for at least one month. The aptamer probe (CEAA) self-assembled on the nanorods' surface via an Au–S bond, creating the magnetic probe (CEAA/α-Fe2O3/Fe3O4@Au). Upon capturing CEA, the current response decreased, indicating a “turn-off" nano-biosensor type as observed through differential pulse voltammetry (DPV) monitoring. In this research, the biosensor not only exhibited robust specificity and superior anti-interference ability but also demonstrated successful regeneration. The limit of detection (LOD) was 0.197 pg/mL, the recovery rate for real serum samples was between 94.15 % and 105.08 %, and the relative standard deviation (RSD) ranged from 1.40 % to 2.09 %. This biosensor showed promise in enhancing the accuracy and effectiveness of CRC diagnostic and prognostic protocols, offering a new direction for point-of-care testing.

Label-free electrochemical biosensor based on dual amplification of gold nanoparticles and polycaprolactones for CEA detection

Carcinoembryonic Antigen (CEA), an acidic glycoprotein with human embryonic antigen properties, is found on the surface of cancer cells that have differentiated from endodermal cells. This paper presents a label-free electrochemical immunoassay for the dual amplification detection of CEA using gold nanoparticles loaded with polypyrrole polydopamine (Au/PPy-PDA) and polymerized polycaprolactone (Ng-PCL) prepared by ring-opening polymerization (ROP). First, the composite Au/PPy-PDA was adhered to the electrode surface. Then, gold nanoparticles form a Au–S bond with the sulfhydryl group in Apt1 to secure it on the electrode surface. Subsequently, the non-specific binding sites on the electrodes surface are closed by bovine serum albumin (BSA). Next, CEA is dropped onto the electrode surface, which is immobilized by antigen-antibody specific recognition, and the carboxyl-functionalized Apt2 forms a "sandwich structure" of antibody-antigen-antibody by specific recognition. Polymeric Ng-PCL is adhered to the electrode surface, leading to an increase in the electrochemical impedance signal, resulting in a complete chain of signal analysis. Finally, the response signal is detected by electrochemical impedance spectroscopy (EIS). Under optimal experimental conditions, the method has the advantages of high sensitivity and wide linear range (1 pg mL−1∼100 ng mL−1), and the lower limit of detection (LOD) is 0.234 pg mL−1. And it has the same high sensitivity, selectivity and interference resistance for the real samples detection. Thus, it provides a new way of thinking about biomedical and clinical diagnosis.

Detection of Ovarian Cancer Biomarker Lysophosphatidic Acid Using a Label-Free Electrochemical Biosensor

Electrochemical biosensors are valued for their sensitivity and selectivity in detecting biological molecules. Having the advantage of generating signals that can be directly or indirectly proportional to the concentration of the target analyte, these biosensors can achieve specificity by utilizing a specific biorecognition surface designed to recognize the target molecule. Electrochemical biosensors have garnered substantial attention, as they can be used to fabricate compact, cost-effective devices, making them promising candidates for point-of-care testing (POCT) devices. This study introduces a label-free electrochemical biosensor employing a gold screen-printed electrode (SPE) to detect lysophosphatidic acid (LPA), a potential early ovarian cancer biomarker. We employed the gelsolin–actin system, previously introduced by our group, in combination with fluorescence spectrometry, as a biorecognition element to detect LPA. By immobilizing a gelsolin–actin complex on an SPE, we were able to quantify changes in current intensity using cyclic voltammetry and differential pulse voltammetry, which was directly proportional to the LPA concentration in the solution. Our results demonstrate the high sensitivity of the developed biosensor for detecting LPA in goat serum, with a limit of detection (LOD) and a limit of quantification (LOQ) of 0.9 µM and 2.76 µM, respectively, highlighting its potential as a promising tool for early-stage diagnosis of ovarian cancer.

Target-promoted autocatalytic hairpin assembly of bivalent DNAzymes for sensitive and label-free electrochemical metallothionein assay

Metallothionein (MT) has shown to be an important biomarker for environmental monitoring and various diseases, due to its significant binding ability to heavy metal ions. On the basis of such a characteristic and the Hg2+-stabilized DNA duplex (Hg2+-dsDNA) probe, as well as a new autocatalytic hairpin assembly (aCHA)/DNAzyme cascaded signal enhancement strategy, the construction of a highly sensitive and label-free electrochemical MT biosensor is described. Target MT molecules bind Hg2+ in Hg2+-dsDNA to disrupt the duplex structure and to release ssDNA sequences, which trigger subsequent aCHA for efficient production of mimic aCHA triggering strands and many bivalent DNAzymes. The signal hairpins on the electrode are then cyclically cleaved by DNAzyme amplification cascade to liberate plenty G-quadruplex sequences, which bind hemin and yield largely enhanced currents for sensitive assay of MT with a detection limit of 0.217 nM in a label-free approach. Such sensor also shows selective discrimination capability to MT against other interfering proteins and assay of MT in normal serums with dilution has also been verified, indicating its potential for highly sensitive detection of different heavy metal ion binding molecules for various application scenarios.

Constructing a label-free electrochemical biosensor based on magnetic α-Fe2O3/Fe3O4 heterogeneous nanosheets for the sensitive detection of VKORC1*2 gene

Pharmacogenetic testing technology can effectively determine the individual differences among patients and scientifically assist doctors to select the most suitable drugs for each patient. In this study, an electrochemical biosensor based on magnetic α-Fe2O3/Fe3O4 heterogeneous nanosheets was constructed to detect Vitamin K epoxide reductase complex 1 type AA (VKORC1*2). Firstly, we fabricated magnetic α-Fe2O3/Fe3O4 heterogeneous nanosheets as the substrate material for electrochemical biosensors. Subsequently, the material surface was modified with Au nanoparticles to facilitate the connection of sensing probes and enhance current signal amplification. The electrochemical biosensor showed a negative linear relationship with the concentrations of target DNA (tDNA, VKORC1*2 gene) within 1 pM − 1 μM, a limit of detection was (LOD) = 0.36 pM, and a limit of quantification was (LOQ) = 1.19 pM. The biosensor demonstrated exceptional specificity, reproducibility, and stability. In real sample analysis, it exhibited a recovery range of 96.63 % − 110.57 % (RSD ≤ 3.07 %) for various tDNA concentrations, thereby indicating its promising potential in clinical diagnostics.

Development of label-free electrochemical OMP-DNA probe biosensor as a highly sensitive system to detect of citrus huanglongbing

The fabrication of the first label-free electrochemical DNA probe biosensor for highly sensitive detection of Candidatus Liberibacter asiaticus (CLas), as the causal agent of citrus huanglongbing disease, is conducted here. An OMP probe was designed based on the hybridization with its target-specific sequence in the outer membrane protein (OMP) gene of CLas. The characterization of the steps of biosensor fabrication and hybridization process between the immobilized OMP-DNA probe and the target ssDNA oligonucleotides (OMP-complementary and three mismatches OMP or OMP-mutation) was monitored using cyclic voltammetry and electrochemical impedance spectroscopy based on increasing or decreasing in the electron transfer in [Fe (CN)6]3−/4− on the modified gold electrode surface. The biosensor sensitivity indicated that the peak currents were linear over ranges from 20 to 100 nM for OMP-complementary with the detection limit of 0.026 nM (S/N = 3). The absence of any cross-interference with other biological DNA sequences confirmed a high selectivity of fabricated biosensor. Likewise, it showed good specificity in discriminating the mutation oligonucleotides from complementary target DNAs. The functional performance of optimized biosensor was achieved via the hybridization of OMP-DNA probe with extracted DNA from citrus plant infected with CLas. Therefore, fabricated biosensor indicates promise for sensitivity and early detection of citrus huanglongbing disease.

Label-free electrochemical biosensor based on green-synthesized reduced graphene oxide/Fe3O4/nafion/polyaniline for ultrasensitive detection of SKBR3 cell line of HER2 breast cancer biomarker

Cancer stands as one of the most impactful illnesses in the modern world, primarily owing to its lethal consequences. The fundamental concern in this context likely stems from delayed diagnoses in patients. Hence, detecting various forms of cancer is imperative. A formidable challenge in cancer research has been the diagnosis and treatment of this disease. Early cancer diagnosis is crucial, as it significantly influences subsequent therapeutic steps. Despite substantial scientific efforts, accurately and swiftly diagnosing cancer remains a formidable challenge. It is well known that the field of cancer diagnosis has effectively included electrochemical approaches. Combining the remarkable selectivity of biosensing components—such as aptamers, antibodies, or nucleic acids—with electrochemical sensor systems has shown positive outcomes. In this study, we adapt a novel electrochemical biosensor for cancer detection. This biosensor, based on a glassy carbon electrode, incorporates a nanocomposite of reduced graphene oxide/Fe3O4/Nafion/polyaniline. We elucidated the modification process using SEM, TEM, FTIR, RAMAN, VSM, and electrochemical methods. To optimize the experimental conditions and monitor the immobilization processes, electrochemical techniques such as CV, EIS, and SWV were employed. The calibration graph has a linear range of 102–106 cells mL−1, with a detection limit of 5 cells mL−1.

Combining WO3@AuNPs with Poly(amidoamine) Allows Sensitive Electrochemical Detection of DR1 Based on Dual Signal Amplification.

Down-regulator of transcription 1 (DR1) is considered as a biomarker of hashimoto's thyroiditis (HT), which is a risk factor for thyroid cancer. Here, a label-free electrochemical biosensor for DR1 detection was constructed based on polyamidoamine (PAMAM) polymer and the nanocomposite (WO3@AuNPs) composed of tungsten trioxide (WO3) and gold nanoparticles (AuNPs). WO3@AuNPs was obtained by combining monolayer WO3 nanosheets, which has high conductivity, and AuNPs. The modification of WO3@AuNPs can not only increase the conductivity of the electrode but also provide more active sites for signaling units, thus greatly improve the sensitivity of the sensor. The polymer PAMAM is biocompatible and non-immunogenic, and its end functional group can bind to the target molecules, providing them with more binding sites and thus improving the sensitivity of the sensor. Under optimal conditions, the label-free biosensor showed a good linear relationship between the logarithm of DR1 concentration and the impedance in the range of 10 fg ⋅ mL-1 to 100 ng ⋅ mL-1, with a detection limit as low as 0.3 fg ⋅ mL-1. Besides, this label-free electrochemical platform exhibited satisfactory selectivity and anti-interference capability in human serum samples. Therefore, this method has considerable potential in clinical detection of DR1.

Immobilization-free and label-free electrochemical DNA biosensing based on target-stimulated release of redox reporter and its catalytic redox recycling

Herein, we demonstrate a simple, homogenous and label-free electrochemical biosensing system for sensitive nucleic acid detection based on target-responsive porous materials and nuclease-triggered target recycling amplification. The Fe(CN)63− reporter was firstly sealed into the pores of Fe3O4 nanoparticles by probe DNA. Target DNA recognition triggered the controllable release of Fe(CN)63− for the redox reaction with the electron mediator of methylene blue enriched in the dodecanethiol assembled electrode and thereby generating electrochemical signal. The exonuclease III (Exo III)-assisted target recycling and the catalytic redox recycling between Fe(CN)63− and methylene blue contributed for the enhanced signal response toward target recognition. The low detection limit toward target was obtained as 478 fM and 1.6 pM, respectively, by square wave voltammetry and cyclic voltammetry methods. It also possessed a well-discrimination ability toward mismatched strands and high tolerance to complex sample matrix. The coupling of bio-gated porous nanoparticles, nuclease-assisted target amplification and catalytic redox recycling afforded the sensing system with well-controllable signal responses, sensitive and selective DNA detection, and good stability, reusability and reproducibility. It thus opens a new avenue toward the development of simple but sensitive electrochemical biosensing platform.

Aptamer-based label-free electrochemical biosensors for the diagnosis of sickle cell anemia

Aptamer-based label-free electrochemical biosensors for the diagnosis of sickle cell anemia

High-density aptamer synthesis based on primer exchange reaction for label-free electrochemical biosensing

We report an electrochemical aptamer-based (E-AB) sensor for label-free quantitative detection of thrombin. In-situ synthesis of high-density aptamer replicas on the electrode is accomplished based on primer exchange reaction (PER), by which a DNA primer is continuously extended with the presence of a catalytic hairpin template and DNA polymerase. The PER enables the synthesis of long single-stranded DNA with replicated aptamer sequence for thrombin binding. The PER products on the surface of a gold electrode increase the density of aptamers on a limited electrode area, which can provide more effective binding sites for thrombin and therefore improve the performance of the sensor based on electrochemical impedance spectroscopy. Compared with the conventional E-AB sensor based on a 15-mer aptamer immobilized on the electrode, our method has a lower detection limit of 0.91 pM, and a wider linear range of 1.0 pM to 50 nM. Therefore, we believe our method is promising for label-free electrochemical detection of biomarkers.

Selection of ssDNA aptamers and construction of aptameric electrochemical biosensor for the detection of Giardia intestinalis trophozoite protein

Giardia intestinalis is one of the most widespread intestinal parasites and is considered a major cause of epidemic or sporadic diarrhea worldwide. In this study, we aimed to develop a rapid aptameric diagnostic technique for G. intestinalis infection. First, the SELEX (Systematic Evolution of Ligands by Exponential Enrichment) process generated DNA aptamers specific to a recombinant protein of the parasite's trophozoite. Ten selection rounds were performed; each round, the DNA library was incubated with the target protein conjugated to Sepharose beads. Then, the unbound sequences were removed by washing and the specific sequences were eluted and amplified by Polymerase Chain Reaction (PCR). Two aptamers were selected, and the dissociation constants (Kd), were determined as 2.45 and 16.95 nM, showed their high affinity for the G. intestinalis trophozoite protein. Subsequently, the aptamer sequence T1, which exhibited better affinity, was employed to develop a label-free electrochemical biosensor. A thiolated aptamer was covalently immobilized onto a gold screen-printed electrode (SPGE), and the binding of the targeted protein was monitored using square wave voltammetry (SWV). The developed aptasensor enabled accurate detection of the G. intestinalis recombinant protein within the range of 0.1 pg/mL to 100 ng/mL, with an excellent sensitivity (LOD of 0.35 pg/mL). Moreover, selectivity studies showed a negligible cross-reactivity toward other proteins such as bovine serum albumin, globulin, and G. intestinalis cyst protein.

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).