Sandwich-type Immunosensor Research Articles

AIEgens-based luminescent metal-organic frameworks as novel electrochemiluminescence emitters Integrated with co-reaction amplification strategy for CA15-3 detection

A current research focus in the field of electrochemiluminescence (ECL) is the development of novel high-performance emitters. Herein, we reported the synthesis of a zirconium-based metal–organic framework (Zr-TBAPy-MOF) using the aggregation-induced emission luminogens (AIEgens) 1,3,6,8-tetra(4-carboxyphenyl)pyrene (H4TBAPy) as ligands and Zr6-clusters as nodes. The resulting AIE-active luminescent MOF (AIE-LMOF) exhibited superior ECL properties and was used as an ECL emitter to construct a high-sensitivity ECL immunosensor. Conceptual experiments demonstrated that Zr-TBAPy-MOF showed significantly stronger ECL emission compared to both the monomers and aggregates of H4TBAPy, attributed to the effective restriction of intramolecular motion (RIM). On the sensor substrate, we designed an Au@ZnO/Cu2O nanocomposite as a co-reactant accelerator. This significantly promoted the generation of sulfate radicals (SO4•−) from persulfate (S2O82−) through the sustainable switching of Cu+/Cu2+ oxidation states in copper oxide (Cu2O). Additionally, zinc oxide (ZnO), which had excellent electrochemical properties, further enhanced the catalytic activity of the materials. Leveraging these advantages, we developed a sandwich-type ECL immunosensor for the ultrasensitive detection of carbohydrate antigen 15–3 (CA15-3), demonstrating a wide sensitive range (0.001 − 100 U/mL) and a low detection limit (0.0007 U/mL). Overall, this work paves the way for the development of efficient ECL luminophores with significant potential in the field of immunoassays for the early diagnosis of disease.

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.

Enzymatic Precipitation of Highly Electroactive and Ion-Transporting Prussian Blue for a Sensitive Electrochemical Immunosensor.

Sensitive and/or multiplex electrochemical biosensors often require efficient (bio)catalytic conversion of substrates into insoluble electroactive products. The enzymatic formation and precipitation of coordination polymers under mild conditions offers a promising solution for this purpose. Herein, we report the enzymatic precipitation of Prussian blue (PB), a highly electroactive and ion-transporting coordination polymer, on an immunosensing electrode for application in a sensitive electrochemical immunosensor for detecting thyroid-stimulating hormone (TSH). Five pairs of redox enzymes and their specific reductants were examined to achieve rapid PB precipitation and electrochemical oxidation. Among these pairs, O2-insensitive flavin adenine dinucleotide-dependent glucose dehydrogenase (FAD-GDH) paired with glucose yielded the highest electrochemical signal-to-background (S/B) ratio. FAD-GDH catalyzed the conversion of Fe(CN)63- to Fe(CN)64-, which coordinated with Fe3+, leading to PB formation and subsequent precipitation through repeated conversions. The resulting PB precipitate, with its close proximity to the electrode, facilitated rapid electrochemical oxidation and generated a strong electrochemical signal. Notably, the precipitation and electrochemical oxidation of PB were more effective than those of its analogues. When applied to a sandwich-type immunosensor for TSH detection, the enzymatic PB precipitation achieved a calculated detection limit of approximately 2 pg/mL in artificial serum, covering the clinically relevant range. These findings indicate the potential widespread utility of PB precipitation and electrochemical oxidation for sensitive multiplex biomarker detection.

FeNi-MIL-88B-based electrochemiluminescence immunosensor for ultra-sensitive detection of CD44 protein via dual-quenching strategy

BackgroundCluster of Differentiation 44 (CD44) is considered an important biomarker for various cancers, and achieving highly sensitive detection of CD44 is crucial, which plays a significant role in tumor invasion and metastasis, providing essential information for clinical tumor diagnosis. Commonly used methods for analysis include fluorescence spectroscopy (FL), photoelectrochemical analysis (PEC), electrochemical analysis (EC), and commercial ELISA kits. Although these methods offer high sensitivity, they can be relatively complex to perform experimentally. Electrochemiluminescence (ECL) has gained widespread research attention due to its high sensitivity, ease of operation, effective spatiotemporal control, and close to zero background signal. ResultsIn this work, a sandwich-type ECL immunosensor for detecting CD44 was constructed using luminol as a luminophore. In this sensing platform, bimetallic MOFs (Pd@FeNi-MIL-88B) loaded with palladium nanoparticles (Pd NPs) were used as a novel enzyme mimic, exhibiting excellent catalytic performance towards the electroreduction of H2O2. The hybrids provided a strong support platform for luminol and antibodies, significantly enhancing the initial ECL signal of luminol. Subsequently, core-shell Au@MnO2 nanocomposites were synthesised by gold nanoparticles (Au NPs) encapsulated in manganese dioxide (MnO2) thin layers, as labels. In the luminol/H2O2 system, Au@MnO2 exhibited strong light absorption in the broad UV–vis spectrum, similar to the black body effect, and the scavenging effect of Mn2+ on O2•−, which achieved the dual-quenching of ECL signal. Under the optimal experimental conditions, the immunosensor demonstrated a detection range of 0.1 pg mL−1 − 100 ng mL−1, with a detection limit of 0.069 pg mL−1. SignificanceBased on Pd@FeNi-MIL-88B nanoenzymes and Au@MnO2 nanocomposites, a dual-quenching sandwich-type ECL immunosensor for the detection of CD44 was constructed. The proposed immunosensor exhibited excellent reproducibility, stability, selectivity, and sensitivity, and provided a valuable analytical strategy and technical platform for the accurate detection of disease biomarkers, and opened up potential application prospects for early clinical treatment.

‘Three Birds With One Stone’ functionalized metal–organic frameworks for facile and efficient chemiluminescent immunoassays

The luminol–H2O2 system is crucial in the field of chemiluminescence (CL) analysis. However, luminol exhibits improved solubility and luminescence only in an alkaline environment, and its preparation process is cumbersome. This inconvenience highlights the need for developing a simple and universally applicable method in this regard. This study integrates luminol and a catalyst into metal–organic frameworks (MOFs) for fabrication of functionalised MOF nanoparticles. Further, PEGylated hematin (PEG = polyethylene glycol) was prepared. This modification with PEG enhanced the catalytic properties of hematin and facilitated the formation of zeolitic imidazole framework-8 nanoparticles. Furthermore, the built-in catalyst enhanced the CL in terms of strength and persistence. These MOF nanoparticles were used as signal markers to fabricate a sandwich-type CL immunosensor for detection of carcinoembryonic antigen, which is a disease marker, following which highly satisfactory analytical test results are obtained. This approach of integrating luminophores and catalysts offers a new idea for generating novel materials in the CL analysis field.

Europium Metal-Organic Framework with a Tetraphenylethylene-Based Ligand: A Dual-Mechanism Quenching Immunosensor for Enhanced Electrochemiluminescence via the Coordination Trigger.

The effective applications of electrochemiluminescence (ECL) across various fields necessitate ongoing research into novel luminophores and ECL strategies. In this study, self-luminous flower-like nanocomposites (Eu-tcbpe-MOF) were prepared by coordination self-assembly using the aggregation-induced emission material 1,1,2,2-tetrakis(4-carboxyphenyl)ethylene (H4TCBPE) and Eu(III) ions as the precursors. Compared with the monomers and aggregates of H4TCBPE, Eu-tcbpe-MOF exhibits stronger ECL emission. Such enhanced electrochemiluminescence is due to coordination as the coordination-triggered electrochemiluminescence (CT-ECL) enhancement effect. In this study, a cubic-structured nanocomposite (Co9S8@Au@MoS2) was used as an efficient quencher, and a more sensitive ECL detection platform was achieved by two quenching mechanisms: resonance energy transfer and competitive consumption of coreactants. N,N-Diethylethanolamine (DBAE) was used as a coreactant, and DBAE has a faster electron transfer rate and stronger energy supply efficiency than the traditional anodoluminescent coreactant tripropylamine, which effectively improves the ECL signal intensity of Eu-tcbpe-MOF. Hence, a sandwich-type ECL immunosensor was prepared by employing a dual-quenching mechanism, utilizing Eu-tcbpe-MOF as the detection probe and Co9S8@Au@MoS2 as the quencher, achieving precise detection of carcinoembryonic antigen from 0.1 pg·mL-1 to 100 ng·mL-1 with a detection limit of 35.1 fg·mL-1.

Ultrasensitive Upconversion Nanoparticle Immunoassay for Human Serum Cardiac Troponin I Detection Achieved with Resonant Waveguide Grating.

Selective detection of biomarkers at low concentrations in blood is crucial for the clinical diagnosis of many diseases but remains challenging. In this work, we aimed to develop an ultrasensitive immunoassay that can detect biomarkers in serum with an attomolar limit of detection (LOD). We proposed a sandwich-type heterogeneous immunosensor in a 3 × 3 well array format by integrating a resonant waveguide grating (RWG) substrate with upconversion nanoparticles (UCNPs). UCNPs were used to label a target biomarker captured by capture antibody molecules immobilized on the surface of the RWG substrate, and the RWG substrate was used to enhance the upconversion luminescence (UCL) of UCNPs through excitation resonance. The LOD of the immunosensor was greatly reduced due to the increased UCL of UCNPs and the reduction of nonspecific adsorption of detection antibody-conjugated UCNPs on the RWG substrate surface by coating the RWG substrate surface with a carboxymethyl dextran layer. The immunosensor exhibited an extremely low LOD [0.24 fg/mL (9.1 aM)] and wide detection range (1 fg/mL to 100 pg/mL) in the detection of cardiac troponin I (cTnI). The cTnI concentrations in human serum samples collected at different times during cyclophosphamide, epirubicin, and 5-fluorouracil (CEF) chemotherapy in a breast cancer patient were measured by an immunosensor, and the results showed that the CEF chemotherapy did cause cardiotoxicity in the patient. Having a higher number of wells in such an array-based biosensor, the sensor can be developed as a high-throughput diagnostic tool for clinically important biomarkers.

Simple and sensitive sandwich-like voltammetric immunosensing of procalcitonin.

Procalcitonin (PCT) is a reliable biomarker in the early diagnosis of septicemia, pyemia and stroke-associated pneumonia. In this work, through preparing β-cyclodextrin/graphene (CD/GN) nanohybrid as carrier and amplifier simultaneously to band antibodies and probe molecules, a simple and innovative sandwich-like voltammetric immunosensor was proposed for the sensitive and effective determination of PCT. Owing to the host-guest recognition property, the antibodies of PCT can enter into the CD cavities to generate a stable complex; meanwhile, aminopyrene (AP) were introduced as the signal probe and it was adsorbed on the surface of GN via aminopyrine π-πinteraction. Based on the signal change from AP as a response signal which exhibits linearity to the concentration of PCT, a highly sensitive sandwich-type voltammetric immunosensor was developed successfully after optimizing various key parameters. The results demonstrated that the developed sensor had a considerably low detection limit (0.003pgmL-1) and wide linearity of 0.01pgmL-1 to 20.0ngmL-1. This work offered a very simple and sensitive sensing strategy for PCT and other biomarkers via altering the specific antibodies simply, showing great potential 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.

Sandwich-type immunosensor based on aminated 3D-rGOF-NH2 and CMK-3-Fc-MgAl-LDH multilayer nanocomposites for detection of CA125

Cancer antigen 125 (CA125)11Cancer antigen 125 (CA125). is the most important biological screening indicator used to monitor epithelial ovarian cancers, and it plays a vital role in distinguishing ovarian cancers from benign diseases. Biosensors show great potential in the analysis and detection of disease markers. In this study, we designed electrochemical sensors based on three-dimensional amino-functionalized reduced graphene oxide (3D-rGOF-NH2),22three-dimensional amino-functionalized reduced graphene oxide (3D-rGOF-NH2). MgAl layered double hydroxide nanocomposites containing ordered mesoporous carbon (CMK-3),33ordered mesoporous carbon (CMK-3). and ferrocene carboxylic acids（Fc-COOH）44ferrocene carboxylic acids（Fc-COOH）.for the detection of CA125. 3D-rGOF-NH2 possesses good conductivity, a large surface area, and high porosity, enabling more immobilized nanoparticles to be deposited on its surface with excellent stability. CMK-3@Fc@MgAl-LDH nanocomposite was used as a carrier to enhance the immobilization of antibodies and the loading of Fc, conductors to enhance conductivity, and enhancers to gradually amplify the signal of Fc.The surface morphology, elemental composition, and surface groups of the materials were characterized using scanning electron microscopy (SEM),55scanning electron microscopy (SEM). transmission electron microscopy (TEM),66transmission electron microscopy (TEM). and X-ray diffraction (XRD)77X-ray diffraction (XRD). techniques. The response signal of the electrochemical sensor was measured by DPV. Under the optimal conditions, the electrochemical sensor obtained a linear detection range of 0.01 U/mL-100 U/mL with a detection limit of 0.00417 U/mL.

Triple signal-enhanced electrochemiluminescence strategy using iron-based metal-organic frameworks modified with Ru(II) complexes for carcino-embryonic antigen detection

The development of a highly efficient electrochemiluminescence (ECL) emitter represents an effective strategy for enhancing the sensitivity and repeatability of ECL immunosensors. In this study, a sandwich-type ECL immunosensor with triple enhancement was developed to detect carcino-embryonic antigen (CEA). This sensor is based on a porous structure of iron-based metal-organic framework (NH2-MIL-88(Fe)), encapsulating the luminescent tris(2,2′-bipyridine)ruthenium (II) (Ru (bpy)32+), Au@MoS2 with a 3D nanoflower structure as an enhanced substrate. In this system, the MOFs framework encapsulated luminophore was realized to solve its water solubility to reach stable luminescence, as well as the triple enhancement effect based on the principle of amino catalysis, Mo4+/Mo6+ active site conversion, and gold nanoparticles (Au NPs) promotion, which significantly enhanced the detection sensitivity. Furthermore, the ECL immunosensor demonstrated successful application in the highly sensitive and selective detection of CEA, achieving a detection limit of 38.9 fg mL−1. The sensor demonstrates remarkable sensitivity, specificity, stability, repeatability, and practicality in the analysis of human serum samples. This investigation presents a highly effective approach for the ultrasensitive detection of trace proteins.

A signal-off photoelectrochemical sandwich-type immunosensor based on WO3/TiO2 Z-scheme heterojunction.

A sandwich "signal-off" type photoelectrochemical (PEC) immunosensor was fabricated based on a composite heterojunction of tungsten oxide/titanium oxide microspheres (WO3/TiO2) acting as signal amplification platform and carbon microspheres loaded by gold nanoparticles (Cs@Au NPs) utilized as the label for detecting antibody. WO3/TiO2 had excellent photoelectric performance, and the results of Mott-Schottky plots, open-circuit voltage, and electron spin resonance spectroscopy indicated that it belonged to the Z-scheme heterojunction transfer mechanism of photogenerated carriers. To achieve the sensitization of PEC immunosensor, Cs@Au NP-labeled immunocomplex can effectively reduce the photocurrent signal. The PEC immunosensors were fabricated under the optimal conditions of 1:1 WO3/TiO2 (molar ratio), 2.0mgmL-1 WO3/TiO2, and 1.5mgmL-1 Cs@Au NPs. Through comparison of the detection results of label-free and sandwich-type PEC immunosensors for nucleocapsid (N) protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), we found that the sensitivity of the sandwich type was 2.53 times the label-free type, and the limit of detection was 0.006ngmL-1, i.e., 3.17 times lower than the label-free type. This demonstrates that the developed sandwich-type PEC immunosensor will have a brighter application prospect.

Dual Signal-Enhanced Electrochemiluminescence Strategy Based on Functionalized Biochar for Detecting Aflatoxin B1.

Metal-organic frameworks (MOFs) are often used as carriers in the preparation of electrochemiluminescent (ECL) materials, and ECL materials stabilized in the aqueous phase can be prepared by encapsulating chromophores inside MOFs by an in situ growth method. In this study, nanocomposites MIL-88B(Fe)-NH2@Ru(py)32+ with excellent ECL response were prepared by encapsulating Tris(2,2'-bipyridine)ruthenium dichloride (Ru(py)32+) inside MIL-88B(Fe)-NH2 using the one-step hydrothermal method. MIL-88B(Fe)-NH2 possesses abundant amino groups, which can accelerate the catalytic activation process of K2S2O8, and its abundant pores are also conducive to the enhancement of the transmission rate of co-reactant agents, ions, and electrons, which effectively improves the ECL efficiency. In order to obtain more excellent ECL signals, we prepared aminated biochar (NH2-biochar) using Pu-erh tea dregs as precursor and loaded gold nanoparticles (Au NPs) on its surface as substrate material for modified electrodes. Both NH2-biochar and Au NPs can also be used as a co-reactant promoter to catalyze the activation process of co-reactant K2S2O8. Therefore, a sandwich-type ECL immunosensor was prepared based on a dual signal-enhanced strategy for the highly sensitive and selective detection of aflatoxin B1 (AFB1). Under the optimal experimental conditions, the sensitive detection of AFB1 was achieved in the range of 1 pg·mL-1~100 ng·mL-1 with a detection limit of 209 fg·mL-1. The proposed dual signal-enhanced ECL immunosensor can provide a simple, convenient, and efficient method for the sensitive detection of AFB1 in food and agricultural products.

NIR-responsive photoelectrochemical sensing platform for the simultaneous determination of tetrodotoxin and okadaic acid in Nassariidae

Simultaneous detection of tetrodotoxin (TTX) and okadaic acid (OA) is important for seafood safety. In this work, a novel paper electrode-based near-infrared (NIR) light-responsive photoelectrochemical (PEC) immunosensor was constructed using Ag2S quantum dots (QDs) and NaYF4: Yb, Er upconversion nanoparticles (UCNPs) matched with BiOI for the simultaneous detection of TTX and OA in aquatic products. A low-cost, easily prepared gold nanoparticle-functionalized paper-based screen-printed electrode with six channels was designed to immobilize OA and Ab1 of TTX. Correspondingly, PEC signal immunoprobes (BiOI@UCNPs-Ab and Ab2-Ag2S QDs) with NIR-light response were introduced to construct competitive-type and sandwich-type PEC immunosensors for OA and TTX, respectively. Under optimal conditions, the linear ranges for TTX and OA were 0.001–100 and 0.001–80 ng mL−1, respectively, and the detection limits were 5 and 7 pg mL−1, respectively. The proposed sensor was successfully used for the simultaneous analysis of TTX and OA in Nassariidae samples.

Enhancing the Electrochemiluminescence of Porphyrin via Crystalline Networks of Metal-Organic Frameworks for Sensitive Detection of Cardiac Troponin I.

Porphyrins easily aggregate due to unfavorable π-π accumulation, causing luminescent quenching in the aqueous phase and subsequently reducing luminescent efficiency. It is a feasible way to immobilize porphyrin molecules through metal-organic framework materials (MOFs). In this study, 5,10,15,20-tetrakis (4-carboxyphenyl) porphyrin (TCPP) was introduced into the metal-organic skeleton (PCN-224) as a ligand. The result showed that the electrochemiluminescence (ECL) and photoluminescence (PL) efficiency of the MOF skeleton was 8.2 and 6.5 times higher than TCPP, respectively. Impressively, the periodic distribution of porphyrin molecules in the MOF framework can overcome the bottleneck of porphyrin aggregation, resulting in the organic ligand TCPP participating in the electron transfer reaction. Herein, based on the PCN-224, a sandwich-type ECL immunosensor was constructed for the determination of cardiac troponin I (cTnI). It provided sensitive detection of cTnI in the range of 1 fg/mL to 10 ng/mL with a detection limit of 0.34 fg/mL. This work not only innovatively exploited a disaggregation ECL (DIECL) strategy via the crystalline framework of MOF to enhance the PL and ECL efficiency of porphyrin but also provided a promising ECL platform for the ultrasensitive monitoring of cTnI.

Two-dimensional substrate assisted SERS immunosensor for accurate detection of carcinoembryonic antigen

Accurate and sensitive detection of carcinoembryonic antigen (CEA) is essential for the detection of various diseases in healthcare and the medical field. Currently, due to the high false negative rate of CEA assay in clinical setting and its use as a common indicator for early cancer screening, a novel CEA detection method with high sensitivity, increased specificity and the lower cost has become a clinical challenge. Here, a facile sandwich type immunosensor based on surface-enhanced Raman scattering (SERS) was presented including 4-mercaptobenzonitrile (4MBN) labeled gold core-silver shell nanoparticles (Au@4MBN@Ag NPs) as SERS tag and 4-mercaptophenylboronic acid (4-MPBA) functionalized two-dimensional (2D) silver nanoparticle film (Ag FM) as SERS capture substrate for CEA detection. A linearity of 10-9-10-14M was observed with high sensitivity and excellent selectivity for the detection of CEA. Additionally, the spiking experiment yielded 105.33–127.00% recovery with variation coefficients below 10% demonstrating high assay accuracy and precision. The immunosensor we proposed here is a promising approach to quantify CEA in liquid biopsy samples with high sensitivity, which could be further developed for early cancer screening.

Electrochemiluminescence resonance energy transfer between Ru(bpy)32+@Cu3(HHTP)2 and GO-Au composites for C-reactive protein detection

Designing innovative electrochemiluminescence (ECL) immunosensors is critical for the detection of biomarkers with a low concentration and the precise evaluation of clinical diseases. Herein, a Cu3(hexahydroxytriphenylene)2 (Cu3(HHTP)2) nanoflake-based sandwich-type ECL immunosensor was constructed for C-Reactive Protein (CRP) detection. The Cu3(HHTP)2 nanoflake, an electronically conductive metal–organic framework (MOF), has a periodically arranged porous structure with a cavity size of 2 nm, which not only accommodates a large amount of Ru(bpy)32+ but also confines the spatial diffusion of active species. Therefore, the Ru(bpy)32+-loaded Cu3(HHTP)2 nanocomplex (Ru@CuMOF) as an ECL emitter exhibits an enhanced ECL efficiency. The ECL resonance energy transfer (ECL-RET) was accomplished by combining Ru@CuMOF used as a donor with gold nanoparticles-functionalized graphene oxide nanosheets (GO-Au) utilized as an acceptor. This should be ascribed to the fact that the ECL emission spectrum of Ru@CuMOF shows the strongest signal intensity at 615 nm, overlapping with the absorption spectrum of GO-Au at 580–680 nm. Targeted detection of CRP in human serum samples was achieved by the sandwich-type immunosensor based on the ECL-RET mechanism with a 0.26 pg mL−1 detection limit. The electro-activated hybrids of Cu3(HHTP)2 and ECL emitters provide a new sensing strategy for the high-sensitivity detection of disease markers.

Biosensing of telomerase antigen using sandwich type immunosensor based on poly(β-Cyclodextrin) decorated by Au@Pt nanoparticles: An innovative immune-platform toward early-stage identification of cancer

Telomeres maintain genomic integrity in normal cells and their gradual shortening during successive cell divisions causes chromosomal instability. Sensitive and reliable analysis of telomerase activity is essential for clinical diagnosis of cancer. In this study, a novel sandwich type and label-free electrochemical immunosensor was developed for the efficient monitoring of telomerase antigen based on Au@Pt nano-alloy and Poly (beta-Cyclodextrin (β-CD)) functionalized with AuNPs-DDT and toluidine blue. For the first time, KCC-1-NH-CS2 was used as an advanced substrate to encapsulation of biotinylated antibody (Ab1) of telomerase towards maintain of antibody in the stress condition of solution. By providing a wide surface to volume using dendritic fibrous Nanosilica for the effective immobilization of the biotinylated antibody, a sensitive immunosensor was designed for the probing of antigen–antibody immunocomplex. Interestingly, the P(β-CD) assured the bioactivity and stability of the immobilized 2-AG antibody. To complete the capping part of the immunosensor (reporter), an HRP-conjugated antibody of telomerase (secondary antibody (Ab2)) was coupled to the surface of a gold electrode (AuE) modified by P(β-CD), as well as a predominantly biotinylated antibody (Ab1, encapsulated into the KCC-1-NH-CS2). Engineered immune-platform demonstrated acceptable analytical properties (LLOQ = 0.078 IU.ml). In addition, the created biosensor shows excellent selectivity and acceptable ability for the detection of telomerase in human plasma samples with satisfactory stability which might hold great application potential in cancer analysis and clinical diagnosis. This research inspires scientists and researchers to generate and develop novel biosensor platforms for point-of-care (POC) diagnostics of telomerase. Based on accurate, sensitive, inexpensive, and time-saving futures of developed biosensor compared to other methods makes it a useful platform for future analytical works.

An electrochemical immunosensor based on polyaniline microtubules and zinc gallinate for detection of human growth differentiation factor-15.

The incidence rate of cardiovascular diseases (CVDs) remains high, and their mortality rate is significantly higher than that of other diseases. Growth differentiation factor-15 (GDF-15) is a recently developed biomarker for the early diagnosis and prognostic evaluation of CVDs because its concentration in serum increases substantially after a cardiovascular injury or an inflammatory reaction. In this study, a sandwich-type immunosensor was constructed for the sensitive detection of GDF-15. Specifically, peony-like zinc gallinate (ZnGa2O4) prepared using a hydrothermal method, which exhibits excellent electrocatalytic performance, was coupled with Au nanoparticles (NPs) to obtain golden-peony-like ZnGa2O4/Au NPs. Theypreserved the immune activity of GDF-15 antibody molecules and further enhanced the conductivity, thereby realizing additional signal amplification. Hollow polyaniline (PANI) microtubules decorated with Pd NPs were used as the sensing platform (PANI/Pd NPs). The hollow microtubules provided abundant active sites and considerably improved the electron-transfer rate. Under optimal conditions, a linear range and remarkably low detection limit of 100fgmL-1-10ngmL-1 and 42.23fgmL-1, respectively, were achieved. These experimental results indicate that the strategy reported herein can be adopted as a novel approach for the convenient and rapid detection of GDF-15.

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.