Sandwich-type Electrochemical Immunosensor Research Articles

A novel sandwich electrochemical immunosensor utilizing customized template and phosphotungstate catalytic amplification for CD44 detection

A sandwich-type electrochemical immunosensor was proposed for the ultra-sensitive detection of CD44, a potential biomarker for breast cancer. In this design, a customized template-based ionic liquid (1-butyl-2,3-dimethylimidazolium tetrafluoroborate) carbon paste electrode (CILE) served as the sensing platform, and thionine/Au nanoparticles/covalent-organic frameworks (THI/Au/COF) were used as the signal label. Moreover, an enzyme-free signal amplification strategy was introduced by involving H2O2 and phosphotungstate (PW12) with peroxidase-like activity. Under optimized conditions, the linear range is as wide as six orders of magnitude, and the detection limit is as low as 0.71 pg mL−1 (estimated based on S/N = 3). Average recoveries range from 98.16 %-100.1 %, with a relative standard deviation (RSD) of 1.42–8.27 % in mouse serum, and from 98.44 %-99.06 %, with an RSD of 1.14–4.84 % (n = 3) in artificial saliva. Furthermore, the immunosensor exhibits excellent specificity toward CD44, good stability, and low cost, indicating great potential for application in clinical trials.

Cooperative amplification of Prussian blue as a signal indicator and functionalized metal-organic framework-based electrochemical biosensor for an ultrasensitive HE4 assay

Human epididymis protein 4 (HE4), a diagnostic biomarker of ovarian cancer, is crucial for monitoring the early stage of the disease. Hence, it is highly important to develop simple, inexpensive, and user-friendly biosensors for sensitive and quantitative HE4 assays. Herein, a new sandwich-type electrochemical immunosensor based on Prussian blue (PB) as a signal indicator and functionalized metal-organic framework nanocompositesas efficient signal amplifiers was fabricated for quantitative analysis of HE4. In principle, ketjen black (KB) and AuNPs modified on TiMOF (TiMOF-KB@AuNPs) could accelerate electron transfer on the electrode surface and act as a matrix for the immobilization of antibodies via cross-linking to improve the determination sensitivity. The PB that covalently binds to labeled antibodies endows the biosensors with intense electrochemical signals. Furthermore, the concentration of HE4 could be indirectly detected by monitoring the electroactivity of PB. Benefiting from the high signal amplification ability of the PB and MOF nanocomposites, this strategy displayed a wide linear range (0.1–80 ng mL−1) and a lower detection limit (0.02 ng mL−1). Hence, this study demonstrated great promise for application in clinical ovarian cancer diagnosis and treatment, and provided a new platform for detecting other cancer biomarkers.

An ultrasensitive sandwich-type electrochemical immunosensor based on rGO-TEPA/ZIF67@ZIF8/Au and AuPdRu for the detection of tumor markers CA72-4

Cancer antigen 72–4 (CA72-4) is an important marker of cancer detection, and accurate detection of CA72-4 is urgently required. Herein, a sandwich-type immunosensor was constructed for detection CA72-4 based on composite nanomaterial as the substrate material and trimetal nanoparticles as the nanoprobe. The composite nanomaterial rGO-TEPA/ZIF67@ZIF8/Au used as a selective bio-recognition element were modified on the glassy carbon electrode (GCE) surface. Meanwhile, the electrochemical nanoprobes were fabricated through the AuPdRu trimeric metal. After the target antigen 72–4 were captured, the nanoprobes were further assembled to form an antibody1 (Ab1)- antigen-antibody2 (Ab2) nanoprobes sandwich-like system on the electrode surface. Then, hybrid the substrate material rGO-TEPA/ZIF67@ZIF8/Au and the AuPdRu trimeric metal nanoprobes efficiently catalyzed the reduction of H2O2 and amplified the electrochemical signals. Cyclic voltammetry (CV), differential pulse voltammetry (DPV), electrochemical impedance spectroscopy (EIS), and Chronoamperometry (I-T) methods were used to characterize the performance and detection capabilities for CA72-4 of the prepared immunosensors. The results showed that the detection limit was 1.8 × 10−5 U/mL (S/N = 3), and the linear range was 0.001–1000 U/mL. This study provides a new signal amplification strategy for electrochemical sensors and a theoretical basis for the clinical application of immunosensor to detect other tumor markers.

Electrochemical Detection of a Breast Cancer Biomarker with an Amine-Functionalized Nanocomposite Pt-Fe3O4-MWCNTs-NH2 as a Signal-Amplifying Label.

We report an ultrasensitive sandwich-type electrochemical immunosensor to detect the breast cancer biomarker CA 15-3. Amine-functionalized composite of reduced graphene oxide and Fe3O4 nanoparticles (MRGO-NH2) was used as an electrochemical sensing platform material to modify the electrodes. The nanocomposite comprising Pt and Fe3O4 nanoparticles (NPs) anchored on multiwalled carbon nanotubes (Pt-Fe3O4-MWCNTs-NH2) was utilized as a pseudoenzymatic signal-amplifying label. Compared to reduced graphene oxide, the composite MRGO-NH2 platform material demonstrated a higher electrochemical signal. In the Pt-Fe3O4-MWCNTs-NH2 label, multiwalled carbon nanotubes provided the substratum to anchor abundant catalytic Pt and Fe3O4 NPs. The nanocomposites were thoroughly characterized using transmission electron microscopy, Fourier transform infrared spectroscopy, X-ray diffraction, scanning electron microscopy, and X-ray photoelectron spectroscopy. An electroanalytical study and prevalidation of the immunosensor was carried out. The immunosensor exhibited exceptional capabilities in detecting CA 15-3, offering a wider linear range of 0.0005-100 U mL-1 and a lower detection limit of 0.00008 U mL-1. Moreover, the designed immunosensor showed good specificity, reproducibility, and acceptable stability. The sensor was successfully applied to analyze samples from breast cancer patients, yielding reliable results.

An ultrasensitive electrochemical immunosensor based on meso-PdN NCs and Au NPs/N-CNTs for quantitative cTnI detection

Electrochemical immunosensors have gained considerable attention in detecting human disease markers due to their excellent specificity, high sensitivity, and facile operation. Herein, a rational-designed sandwich-type electrochemical immunosensor is constructed for the sensitive detection of cardiac troponin I (cTnI) using nitrogen-doped carbon nanotubes loaded with gold nanoparticles (Au NPs/N-CNTs) as substrate and highly active mesoporous palladium-nitrogen nanocubes (meso-PdN NCs) as secondary antibody markers. Benefitting from its large specific surface area (638.04 m2 g−1) and high nitrogen content, novel polydopamine (PDA)/ halloysite nanotubes (HNTs) hybrid derived one-dimensional (1D) N-CNTs can provide more binding sites for the in-situ growth of Au NPs to connect Ab1. Furthermore, as an ideal substrate material, Au NPs/N-CNTs exhibit finely tuned mesoporous structures and outstanding conductivity, which facilitate the mass and electron transfer during the electrocatalysis process. Besides, highly concave surfaces and crystalline mesopores of meso-PdN NCs expose more surfaces and crevices, providing abundant reactive sites for H2O2 reduction. Remarkably, the as-obtained immunosensor presented a wide linear range (from 10 fg mL−1 to 100 ng mL−1) and an excellent low detection limit (9.85 fg mL−1). This study may offer new insights into the precise fabrication of efficient electrochemical immunosensors for various clinical diagnosis applications.

A sensitive sandwich-type electrochemical immunosensor using nitrogen-doped graphene/metal-organic framework-derived CuMnCoOx and Au/MXene for the detection of breast cancer biomarker

In terms of cancer-related deaths among women, breast cancer (BC) is the most common. Clinically, human epidermal growth receptor 2 (HER2) is one of the most commonly used diagnostic biomarkers for facilitating BC cell proliferation and malignant growth. In this study, a disposable gold electrode (DGE) modified with gold nanoparticle-decorated Ti3C2Tx (Au/MXene) was utilized as a sensing platform to immobilize the capturing antibody (Ab1/Au/MXene). Subsequently, nitrogen-doped graphene (NG) with a metal-organic framework (MOF)-derived copper-manganese-cobalt oxide, tagged as NG/CuMnCoOx, was used as a probe to label the detection antibody (Ab2). A sandwich-type immunosensor (NG/CuMnCoOx/Ab2/HER2-ECD /Ab1/Au/MXene/DGE) was developed to quantify HER2-ECD. NG/CuMnCoOx enhances the conductivity, electrocatalytic active sites, and surface area to immobilize Ab2. In addition, Au/MXene facilitates electron transport and captures more Ab1 on its surface. Under optimal conditions, the resultant immunosensor displayed an excellent linear range of 0.0001 to 50.0 ng. mL−1. The detection limit was 0.757 pg·mL−1 with excellent selectivity, appreciable reproducibility, and high stability. Moreover, the applicability for determining HER2-ECD in human serum samples indicates its ability to monitor tumor markers clinically.

High-performance electrochemical immunosensor based on bimetallic gold/silver functionalized carbon spheres for CYFRA 21-1 detection and information protection.

Bimetallic nanomaterial-based systems have been widely utilized across various fields due to their remarkable expandability and flexibility, including nanomedicine, diagnostics, and molecular information technology. Here, we constructed an electrochemical immunosensor using bimetallic gold/silver functionalized carbon spheres (AuAg@CSs) and mesoporous silica nanoparticles (MSNs) for the sensitive determination of cytokeratin 19 fragment antigen 21-1 (CYFRA 21-1) and ensuring information protection for textual data. The AuAg@CSs demonstrated exceptional catalytic activity towards hydrogen peroxide, generating a significant current signal. The introduction of CYFRA 21-1 facilitated the binding of MSNs, thereby forming a sandwich-type electrochemical immunosensor that resulted in a notable decrease in current. Notably, the detection limit for CYFRA 21-1 was determined to be 31 fg mL-1, accompanied by high selectivity. Furthermore, extensive textual information can be encrypted and concealed within the current responses of the electrochemical nanosensing system. By establishing a threshold, these current signals can be represented as a series of binary strings, which can subsequently be segmented into shorter strings. Through information coding methods, these shorter binary strings can be assembled and decrypted, ultimately merging into meaningful textual content. This study promotes the synthesis and multifunctional application of bimetallic nanomaterials, providing innovative solutions to enhance the sensing sensitivity of electrochemical immunosensors and paving the way for advancements in molecular digitization.

Halloysite nanotubes-loaded conductive polymer as substrate and label material for sensitive detection of amyloid-β protein by electrochemical immunosensor

Amyloid-beta protein (Aβ) is a unique biomarker for Alzheimer's disease (AD). The sandwich-type electrochemical immunosensor, one of the key tools for detecting biomarkers, relies on a high-performance signal amplification approach to enhance its sensitivity. Ni/PdH nanodendrites (Ni/PdH NDs) have increased catalytic activity due to their unique interaction with palladium hydride and their nickel-rich surface, tunable shape and high specific surface area. Modified halloysite nanotubes (mHNT)-loaded with polypyrrole (PPy@mHNT) possess excellent dispersion and a large surface area. This enables the formation of a conductive network to prevent the accumulation of Ni/PdH NDs. Additionally, it exposes more electrocatalytic active centers, effectively amplifying electrical signals. By utilizing Ni/PdH@PPy@mHNT as the labeling material, it shows a consistent and remarkable electrocatalytic activity in H2O2 reduction, leading to signal amplification. The acid-etched HNT coated with polyaniline (PANI@eHNT) exhibits an exceptionally low background signal and outstanding conductivity. This not only accelerates electron transfer on the electrode surface, but also ensures the stable incubation of biomolecules post-amino grafting. Utilizing NH2-PANI@eHNT as a substrate material can guarantee stable biomolecule incubation, offer a stable sensing platform and enhance immunosensor performance. The signal can be amplified and the immunosensor's sensitivity can be raised through the efficient cooperation of the aforementioned nanomaterials. Under optimum circumstances, the electrochemical immunosensor had the lowest detection limit of 5.53 fg mL−1 and a linear range of 50 fg mL−1 to 100 ng mL−1. Based on the outstanding performance previously mentioned, this immunosensor is anticipated to aid in the early detection of AD.

A novel sandwich-type electrochemical immunosensor for sensitive detection of zearalenone using NG/PDDA/HNTs and Ti-MOF-KB composites for signal amplification

In this work, a novel sandwich-type electrochemical immunosensor based on signal amplification was developed for the ultrasensitive detection of zearalenone (ZEA). The composite consisting of poly(diallyldimethylammonium chloride) functionalized nitrogen doped graphene, halloysite nanotubes and toluidine blue (Tb/NG/PDDA/HNTs), was synthesized for the first time. Then it was modified with gold nanoparticles (AuNPs) to bind the secondary antibody (Ab2, 10 μg mL−1) and form the tracer label. In addition, ketjen black (KB) was doped into titanium-based metal–organic framework (Ti-MOF), which provided large specific surface area and employed as the sensing platform to increase the immobilization of the primary antibody (Ab1, 10 μg mL−1). This immunosensor showed a wide linear range for ZEA from 10 fg mL−1 to 100 ng mL−1 with a limit of detection (LOD) as low as 0.57 fg mL−1, which was below the maximum tolerable levels (50∼100 µg kg−1) set by the United Nations Food and Agriculture Organization (FAO).

A sandwich-type electrochemical immunosensor prepared with graphyne-Au triangular nanoprisms and MoS2-AuPtCu hexagonal nanoframes for the detection of human growth/differentiation factor 15

Growth differentiation factor 15 (GDF-15) is an important biomarker for the early diagnosis and prognostic evaluation of cardiovascular disease (CVD). Monitoring the expression level of GDF-15 in serum can enable early diagnosis evaluation of CVDs. Therefore, the realization of high-sensitivity GDF-15 detection platforms is clinically significant. To detect GDF-15, we developed a sandwich-type electrochemical immunosensor based on hydrogen peroxide (H2O2) as a signal source. AuPtCu hexagonal metal nanoframes (AuPtCu HNFs) loaded onto molybdenum disulfide (MoS2) nanosheets (MoS2-AuPtCu HNF) as secondary antibody labels provided signal amplification. The hollow three-dimensional hexagonal-shaped structure of the AuPtCu nanoframe exposed abundant active sites and exhibited excellent catalytic properties. MoS2 nanosheets with flower-like structures and large specific surface areas can effectively load AuPtCu HNFs. The combination of AuPtCu HNFs and MoS2 nanosheets enhanced the catalytic activity toward H2O2 reduction and realized signal amplification. For the substrate material, two-dimensional Au triangular nanoprisms (Au TNPs) with two large surface planes were hybridized with graphyne (GDY) nanosheets. The obtained GDY-Au TNPs exhibited low catalytic activity and excellent electrical conductivity; therefore, they were used to load primary antibodies to effectively enhance the sensitivity. Under optimal conditions, the constructed immunosensor exhibited a wide detection range of 500 fg/mL to 50 ng/mL and low detection limit of 0.212 pg/mL. Moreover, the GDF-15 content in human serum was tested with satisfactory results. Therefore, the constructed immunosensor can sensitively and accurately detect GDF-15 for the early diagnosis of CVD.

Triple signal-enhancing sandwich-type electrochemical immunosensor based on Au@SNHC and PtPdCu/BNC for detection of Cardiac Troponin I

A novel multiple amplification strategy for electrochemical immunoassay was developed elaborately. The realization of this strategy is based on the high efficiency catalysis of boron nitrogen double-doped carbon loaded trimetallic PtPdCu mulberry-like nanospheres (PtPdCu/BNC) and the satisfactory conductivity of gold nanoparticles modified with sulfur nitrogen co-doped hollow porous carbon (Au@SNHC). Single crystal anisotropic Pt octahedral seeds were synthesized with sodium citrate as shape-directing agent, and then three metals were grown in situ to prepare the trimetallic PtPdCu mulberry nanospheres, which had excellent utilization of atoms and a significant number of catalytic active centers. Boron nitrogen double-doped carbon (BNC) with abundant free electrons and high electron density micro-mesoporous can be used as the supported dispersion material of PtPdCu MBNSs, further catalytically enrich H2O2. Moreover, the etching vulcanization of small ZIF template and the co-doping of sulfur and nitrogen endow sulfur nitrogen co-doped hollow porous carbon (SNHC) with shorter the diffusion distance of charge carrier and lower mass transfer resistance. Under optimal conditions, the developed immunosensor exhibits good sensing performance with a detection limit of 4.27 fg⋅mL−1, and the wide linear dynamic range (10 fg⋅mL−1–100 ng⋅mL−1). This study provides a possible strategy for early diagnosis and postoperative monitoring of AMI.

An Electrochemical Immunosensor for Sensitive Detection of Cardiac Troponin I Based on the Amplification Effect of Asymmetric Bowl-Shaped Mesoporous Nanospheres

Here, a sandwich-type electrochemical immunosensor was constructed to detect cardiac troponin I (cTnI) via asymmetric bowl-shaped PdAgFe mesoporous nanospheres (PdAgFe ABMS) as signal amplification label and core–shell cubic Au/Co-LDH@ZIF-67 as substrate material. PdAgFe ABMS prepared by dual-template directional anisotropic island growth method has abundant mesoporous channels to accelerate molecular mass transfer in solution. In particular, the asymmetric bowl-like structure allows more active sites to be exposed, which improves the utilization of atoms to stabilize and high current response signals. The synergistic effect between PdAgFe increased the activation energy of the catalytic reaction and further amplified the current signal. In addition, the biosensing interface based on Au/Co-LDH@ZIF-67 not only exhibits a high electron transfer rate, but also can capture more bioactive molecules. Under the optimal conditions, the constructed immunosensor was detected to exhibit a low limit of detection (LOD, 4.47 fg ml−1) and a wide detection range (10 fg ml−1–100 ng ml−1). This work provides an accurate and convenient scheme for the clinical detection of cTnI.

Sandwich-type electrochemical immunosensor based on CuFe2O4-Pd for cardiac troponin I detection.

A sandwich-type electrochemical immunosensor was designed by highly efficient catalytic cycle amplification strategy of CuFe2O4-Pd for sensitive detection of cardiac troponin I. CuFe2O4 with coupled variable valence metal elements exhibited favorable catalytic performance through bidirectional cycling of Fe2+/Fe3+ and Cu+/Cu2+ redox pairs. More importantly, Cu+ acted as the intermediate product of the catalytic reaction, promoted the regeneration of Fe2+ and ensured the continuous recycling occurrence of the double redox pairs, and significantly amplified the current signal response. Pd nanoparticles (Pd NPs) loaded on the surface of amino-functionalized CuFe2O4 (CuFe2O4-NH2) served as electrochemical mediators to capture labeled antibodies (Ab2), and also as co-catalysts of CuFe2O4 to further enhance the catalytic efficiency, thusimproving the sensitivity of the electrochemical immunosensor. Under the optimal experimental conditions, the linear range was 0.001 ~ 100ng/mL, and the detection limit was 1.91fg/mL. The electrochemical immunosensor has excellent analytical performance, giving a new impetus for the sensitive detection of cTnI.

Pt Nanodot Inlaid Mesoporous NaBiOF Nanoblackberry for Remarkable Signal Amplification Toward Biomarker Detection.

Anew ultrasensitive sandwich-type electrochemical immunosensor has been successfully constructedto quantitatively detect carcinoembryonic antigen (CEA) using blackberry-like mesoporous bismuth-based nanospheres NaBiOF (NBOF NSs) inlaid with Pt nanodots (NDs) (BiPt NSs) as the antibody capture and signal-amplifying probe. The growth of Pt NDs inside the holes of NBOF NSs formed the nanozyme inlay outside NBOF NSs, greatly increasing the specific surface area and exposure of the catalytic active sites by minimizing the particle size of the Pt to nanodot scale. Such a blackberry-shaped heterojunction structure of BiPt NSs was well-suited to antibody capture and improvedthe catalytic performance of BiPt NSs in reducing H2O2, amplifying the signal, and yielding highly sensitive detection of CEA. Theuse of Au nanoparticle-modified multi-walled carbon nanotubes (Au@MWCNTs) as the electrode substrates significantly enhanced the electron transfer behavior over the electrode surface, further increasing the conductivity and sensitivity of the immunosensor. Remarkably, good compatibility with human body fluid was achieved using the newly developed BiPt-based immunosensor resulting from the favorable biocompatibility and stability of both BiPt NSs and Au@MWCNTs. Benefiting from the double signal amplification strategy and the high biocompatibility, the immunosensor responded linearly to CEA in a wide range from 50 fg/mL to 100 ng/ml with an extremely low detection limit of 3.52 fg/mL (S/N = 3). The excellent detection properties of this new immunosensor were evidenced by the satisfactory selectivity, reproducibility, and stability obtained, as well as the reliable and precise determination of CEA in actual human blood samples. This work provides a new strategy for the earlyclinical diagnosis of cancer. Novel blackberry-like mesoporous NaBiOF nanospheres with Pt nanodot inlay were successfully usedto construct a sandwich-type electrochemical immunosensor for the ultra-sensitive detection ofcarcinoembryonic antigen in human blood plasma based on a remarkable signal amplification strategy.

Highly Sensitive Immunosensing of Carcinoembryonic Antigen Based on Gold Nanoparticles Dotted PB@PANI Core-Shell Nanocubes as a Signal Probe.

Herein, a method was developed for the sensitive monitoring of carcinoembryonic antigen (CEA) by gold nanoparticles dotted prussian blue@polyaniline core-shell nanocubes (Au NPs/PB@PANI). First, a facile low-temperature method was used to prepare the uniform PB@PANI core-shell nanocubes with the assistance of PVP, where PB acted as the electron transfer mediator to provide electrochemical signals, and the PANI with excellent conductivity and desirable chemical stability not only played the role of a protective layer to prevent etching of PB in basic media but also effectively improved electron transfer. Importantly, to further enhance the electrical conductivity and biocompatibility of PB@PANI and to further enhance the electrochemical signal and capture a large amount of Ab2, Au NPs were doped on the surface of PB@PANI to form Au NPs/PB@PANI nanocomposites. Subsequently, benefiting from the advantages of core-shell structure nanoprobes and gold-platinum bimetallic nanoflower (AuPt NF), a sandwich-type electrochemical immunosensor for CEA detection was constructed, which provided a wide linear detection range from 1.0 pg·mL-1 to 100.0 ng·mL-1 and a low detection limit of 0.35 pg·mL-1 via DPV (at 3σ). Moreover, it displayed a satisfactory result when the core-shell structure nanoprobe-based immunosensor was applied to determine CEA in real human serum samples.

A sandwich-type electrochemical immunosensor for sensitive detection of HER2 based on dual signal amplification of La-MOF-PbO2 and PEI-MoS2NFs composites

In recent decades, the incidence of breast cancer has increased year by year, posing a serious threat to human health and quality of life, and about 30% of breast cancer patients have human epidermal growth factor receptor 2 (HER2) overexpression. Therefore, HER2 has become an important biomarker and indicator for the clinical evaluation of breast cancer in diagnosis, prognosis and recurrence. In this work, polyethyleneimine functionalized MoS2 nanoflowers (PEI-MoS2NFs) with good electrical conductivity and abundant active binding sites were designed and employed as a sensing platform for immobilizing the primary antibody of HER2 (Ab1). In addition, a La-MOF-PbO2 composite with a large specific surface area and good conductivity was used to load lots of electroactive toluidine blue (TB) and the secondary antibody of HER2 (Ab2) via gold nanoparticles (AuNPs) as the linker. Thus, the constructed sandwich-type electrochemical immunosensor was applied for sensitive detection of HER2, which showed a wide linear range from 100 fg mL−1 to 10 μg mL−1 with a lower limit of detection of 15.64 fg mL−1. Therefore, the resulting immunosensor in this study would have a potential application in clinical bioanalysis.

Sandwich-type electrochemical immunosensing of hypopharyngeal carcinoma biomarker carcinoembryonic antigen based on N-doped hollow mesoporous nanocarbon spheres/gold hybrids as sensing platform and gold/ferrocene as signal amplifier

In the present work, a highly sensitive sandwich-type electrochemical immunosensor of carcinoembryonic antigen (CEA) was developed by preparing N-doped hollow mesoporous nanocarbon spheres/gold hybrids (NHMN/Au) hybridsas sensing platformand Au/ferrocene (Au/Fc) as signal amplifiers. The large surface area and high conductivity as well as good biocompatibility of NHMN/Au can increase the loading of primary antibody (Ab1) and accelerate the electron transport rate of the electrode surface, while Au can carry immobilized secondary antibodies (Ab2) and Fc derivative (Fc-SH).By using Fc-SH as response probe, the experiments show that the peak current of probe could increase after occurring the specific recognition of Ab1-CEA-Ab2, thus a novel sandwich-type immunosensor of CEA was developed. Finally, the proposed method for CEA detection was applied in human serum and the obtained results are satisfactory, indicating the developed method has important clinical applications for CEA determination.

Electrochemical Immunosensor Using Electroactive Carbon Nanohorns for Signal Amplification for the Rapid Detection of Carcinoembryonic Antigen

In this work, a novel sandwich-type electrochemical immunosensor was developed for the quantitative detection of the carcinoembryonic antigen, an important tumor marker in clinical tests. The capture antibodies were immobilized on the surface of a gold disk electrode, while detection antibodies were attached to redox-tagged single-walled carbon nanohorns/thionine/AuNPs. Both types of antibody immobilization were carried out through Au-S bonds using the novel photochemical immobilization technique that ensures control over the orientation of the antibodies. The electroactive SWCNH/Thi/AuNPs nanocomposite worked as a signal tag to carry out both the detection of carcinoembryonic antigen and the amplification of the detection signal. The current response was monitored by differential pulse voltammetry. A clear dependence of the thionine redox peak was observed as a function of the carcinoembryonic antigen concentration. A linear detection range from 0.001-200 ng/mL and a low detection limit of 0.1385 pg/mL were obtained for this immunoassay. The results showed that carbon nanohorns represent a promising matrix for signal amplification in sandwich-type electrochemical immune assays working as a conductive and binding matrix with easy and versatile modification routes to antibody and redox tag immobilization, which possesses great potential for clinical diagnostics of CEA and other biomarkers.

A sandwich-type electrochemical immunosensor based on spherical nucleic acids-templated Ag nanoclusters for ultrasensitive detection of tumor biomarker

The accurate determination of tumor biomarkers in blood is of vital significance in the diagnosis and therapy of tumor disease. In this research, an innovative sandwich-type electrochemical immunosensor is designed for the ultrasensitive determination of tumor biomarker AFP using spherical nucleic acids-templated silver nanoclusters (AgNCs) sensing platform. For this purpose, on one hand, DNA functionalized gold nanoparticles (AuNPs@DNA) is selected not only as the cross-linker to immobilize the primary antibody (anti-AFP antibody 1, Ab1) to obtain AuNPs@DNA-Ab1, but also as the template for synthesizing AgNCs on AuNPs to form AuNPs@DNA-AgNCs. On the other hand, p-sulfonated calix[4]arene (pSC4) modified Au is chosen to immobilize the secondary antibody (anti-AFP antibody 2, Ab2) through host-guest recognition between Ab2 and pSC4. When AFP is encountered, the immunoreaction signal can be significantly amplified by the electrochemical reduction of AgNCs. Under optimal circumstances, the sandwich-type electrochemical immunosensor exhibits broad limit of linearity from 0.001 to 100 ng mL−1 (R2 = 0.997) and low detection limit of 7.74 fg mL−1 (S/N = 3). The immunosensor possesses excellent repeatability and selectivity, offering a novel method for sensitive clinical diagnosis of tumor markers in human hepatocellular carcinoma.

An ultrasensitive electrochemical sensor for detecting porcine epidemic diarrhea virus based on a Prussian blue-reduced graphene oxide modified glassy carbon electrode

This study developed a novel, ultrasensitive sandwich-type electrochemical immunosensor for detecting the porcine epidemic diarrhea virus (PEDV). By electrochemical co-deposition of graphene and Prussian blue, a Prussian blue-reduced graphene oxide-modified glassy carbon electrode was made, further modified with PEDV-monoclonal antibodies (mAbs) to create a new PEDV immunosensor using the double antibody sandwich technique. The electrochemical characteristics of several modified electrodes were investigated using cyclic voltammetry (CV). We optimized the pH levels and scan rate. Additionally, we examined specificity, reproducibility, repeatability, accuracy, and stability. The study indicates that the immunosensor has good performance in the concentration range of 1 × 101.88 to 1 × 105.38 TCID50/mL of PEDV, with a detection limit of 1 × 101.93 TCID50/mL at a signal-to-noise ratio of 3σ. The composite membranes produced via co-deposition of graphene and Prussian blue effectively increased electron transport to the glassy carbon electrode, boosted response signals, and increased the sensitivity, specificity, and stability of the immunosensor. The immunosensor could accurately detect PEDV, with results comparable to real-time quantitative PCR. This technique was applied to PEDV detection and served as a model for developing additional immunosensors for detecting hazardous chemicals and pathogenic microbes.