Mulberry-like porous-hollow AuPtAg nanorods for electrochemical immunosensing of biomarker myoglobin.

Mulberry-like AuPtAg porous hollow nanorods (PHNR) were facilely synthesized for the first time via a wet chemical method, where Au nanorods (Au NR) behaved as sacrificed template. The anisotropic oriented growth and etching process are involved in this synthesis. Their structural and electronic characteristics were scrutinously examined by TEM, EDS, XPS, and electrochemical techniques. The AuPtAg PHNR provided a large specific surface area and exposed a large number of active sites, showing highly enhanced catalytic activity. On this foundation, a label-free electrochemical immunosensor was developed for myoglobin (Myo) assay based on the AuPtAg PHNR. Further, the built sensor exhibited fast and ultrasensitive responses in a linear range of 0.0001 ~ 1000 ng mL-1 with a low limit of detection (LOD = 0.46 pg mL-1, S/N = 3), and enabled efficient application to human serum samples with acceptable results. Consequently, the developed AuPtAg PHNR-based platform has a broad prospect in practically monitoring Myo and other biomarkers in clinics.

Electrochemical Signal Substance for Multiplexed Immunosensing Interface Construction: A Mini Review.

Appropriate labeling method of signal substance is necessary for the construction of multiplexed electrochemical immunosensing interface to enhance the specificity for the diagnosis of cancer. So far, various electrochemical substances, including organic molecules, metal ions, metal nanoparticles, Prussian blue, and other methods for an electrochemical signal generation have been successfully applied in multiplexed biosensor designing. However, few works have been reported on the summary of electrochemical signal substance applied in constructing multiplexed immunosensing interface. Herein, according to the classification of labeled electrochemical signal substance, this review has summarized the recent state-of-art development for the designing of electrochemical immunosensing interface for simultaneous detection of multiple tumor markers. After that, the conclusion and prospects for future applications of electrochemical signal substances in multiplexed immunosensors are also discussed. The current review can provide a comprehensive summary of signal substance selection for workers researched in electrochemical sensors, and further, make contributions for the designing of multiplexed electrochemical immunosensing interface with well signal.

ZnS/C/MoS2 Nanocomposite Derived from Metal-Organic Framework for High-Performance Photo-Electrochemical Immunosensing of Carcinoembryonic Antigen.

A hexafluorophosphate ionic liquid is used as a functional monomer to prepare a metal-organic framework (Zn-MOF). Zn-MOF is used as a template for MoS2 nanosheets synthesis and further carbonized to yield light-responsive ZnS/C/MoS2 nanocomposites. Zn-MOF, carbonized-Zn-MOF, and ZnS/C/MoS2 nanocomposites are characterized by Fourier transform infrared spectroscopy, transmission electron microscopy, X-ray diffraction pattern, scanning electron microscopy (SEM), element mapping, Raman spectroscopy, X-ray photoelectron spectroscopy, fluorescence, and nitrogen-adsorption analysis. Carcinoembryonic antigen (CEA) is selected as a model to construct an immunosensing platform to evaluate the photo-electrochemical (PEC) performances of ZnS/C/MoS2 nanocomposites. A sandwich-type PEC immunosensor is fabricated by immobilizing CEA antibody (Ab1 ) onto the ZnS/C/MoS2 /GCE surface, subsequently binding CEA and the alkaline phosphatase-gold nanoparticle labeled CEA antibody (ALP-Au-Ab2 ). The catalytic conversion of vitamin C magnesium phosphate produces ascorbic acid (AA). Upon being illuminated, AA can react with photogenerated holes from ZnS/C/MoS2 nanocomposites to generate a photocurrent for quantitative assay. Under optimized experimental conditions, the PEC immunosensor exhibits excellent analytical characteristics with a linear range from 2.0 pg mL-1 to 10.0 ng mL-1 and a detection limit of 1.30 pg mL-1 (S/N = 3). The outstanding practicability of this PEC immunosensor is demonstrated by accurate assaying of CEA in clinical serum samples.

An amino-modified metal-organic framework (type UiO-66-NH2) loaded with cadmium(II) and lead(II) ions for simultaneous electrochemical immunosensing of triazophos and thiacloprid.

A method is described for simultaneous voltammetric determination of the pesticides triazophos (TRS) and thiacloprid (THD). A glassy carbon electrode (GCE) was modified with a metal-organic framework (type UiO-66-NH2) which has a large specific surface (1018 m2·g-1) and contains large amounts of Cd(II) and Pb(II) ions, with adsorption capacities of 230 and 271 mg·g-1, respectively. The antigen-loaded particles were then bound to antibody, magnetically separated, and analyzed by square wave voltammetry to give signals for Cd(II) and Pb(II) at -0.82 and - 0.56 V (vs. Ag/AgCl) for TRS and THD, respectively. Under optimized conditions, the method has a wide linear range (0.2-750 ng·mL-1) and low detection limits (0.07 and 0.1 ng·mL-1 at a S/N of 3 for TRS and THD, respectively). It is perceived that this assay represents a useful tool for simultaneous determination of multiple pesticide residues. The method has a wide scope in that may be extended to monitoring of other small organic pollutants by changing the types of metal ions and by using other antibodies. Graphical abstract Schematic presentation of an amino-modified metal-organic framework (type UiO-66-NH2) loaded with Cd(II) and Pb(II) ions for simultaneous electrochemical immunosensing of triazophos (TRS) and thiacloprid (THD). It is based on the fabrication of antigen (Ab)-immobilized UiO-66-NH2-based signal tags (a), and of an antibody (Ab)-immobilized magnetic bead (MB-COOH)-based capture probes (b).

Bio-nanogate manipulation on electrode surface as an electrochemical immunosensing strategy for detecting anti-hepatitis B surface antigen.

The diagnosis of hepatitis B virus (HBV) and monitoring of the vaccination efficiency against HBV require real-time analysis. The presence of antibody against hepatitis B virus surface antigen (anti-HBsAg) as a result of HBV infection and/or immunization may indicate individual immune status towards HBV. This study investigated the ability of a bio-nanogate-based displacement immunosensing strategy in detecting anti-HBsAg antibody, via nonspecific-binding between polyamidoamine dendrimers encapsulated gold nanoparticles (PAMAM-Au) and the 'antigenic determinant' region (aD) of HBsAg. For this purpose, maltose binding protein harbouring the aD region (MBP-aD) was synthesized as a bioreceptor and immobilized on the screen-printed carbon electrode (SPCE). Following that, PAMAM-Au was deposited on MBP-aD, forming the 'gate' and was used as a monitoring agent. Under optimal conditions, the high specificity of anti-HBsAg antibody towards MBP-aD displaced PAMAM-Au causing the decrement of anodic peak in differential pulse voltammetry (DPV) analysis. The signal changes were proportionally related to the concentration of anti-HBsAg antibody, in a range of 1 - 1000 mIU/mL with a limit of detection (LOD) of 2.5 mIU/mL. The results also showed high specificity and selectivity of the immunosensor platform in detecting anti-HBsAg antibody both in spiked buffer and human serum samples.

Highly Stable Buffer-Based Zinc Oxide/Reduced Graphene Oxide Nanosurface Chemistry for Rapid Immunosensing of SARS-CoV-2 Antigens.

The widespread and long-lasting effect of the COVID-19 pandemic has called attention to the significance of technological advances in the rapid diagnosis of SARS-CoV-2 virus. This study reports the use of a highly stable buffer-based zinc oxide/reduced graphene oxide (bbZnO/rGO) nanocomposite coated on carbon screen-printed electrodes for electrochemical immuno-biosensing of SARS-CoV-2 nuelocapsid (N-) protein antigens in spiked and clinical samples. The incorporation of a salt-based (ionic) matrix for uniform dispersion of the nanomixture eliminates multistep nanomaterial synthesis on the surface of the electrode and enables a stable single-step sensor nanocoating. The immuno-biosensor provides a limit of detection of 21 fg/mL over a linear range of 1-10 000 pg/mL and exhibits a sensitivity of 32.07 ohms·mL/pg·mm2 for detection of N-protein in spiked samples. The N-protein biosensor is successful in discriminating positive and negative clinical samples within 15 min, demonstrating its proof of concept used as a COVID-19 rapid antigen test.

Bi-ECDAQ: An electrochemical dual-immuno-biosensor accompanied by a customized bi-potentiostat for clinical detection of SARS-CoV-2 Nucleocapsid proteins.

Multiplex electrochemical biosensors have been used for eliminating the matrix effect in complex bodily fluids or enabling the detection of two or more bioanalytes, overall resulting in more sensitive assays and accurate diagnostics. Many electrochemical biosensors lack reliable and low-cost multiplexing to meet the requirements of point-of-care detection due to either limited functional biosensors for multi-electrode detection or incompatible readout systems. We developed a new dual electrochemical biosensing unit accompanied by a customized potentiostat to address the unmet need for point-of-care multi-electrode electrochemical biosensing. The two-working electrode system was developed using screen-printing of a carboxyl-rich nanomaterial containing ink, with both working electrodes offering active sites for recognition of bioanalytes. The low-cost bi-potentiostat system (∼$80) was developed and customized specifically to the bi-electrode design and used for rapid, repeatable, and accurate measurement of electrochemical impedance spectroscopy signals from the dual biosensor. This binary electrochemical data acquisition (Bi-ECDAQ) system accurately and selectively detected SARS-CoV-2 Nucleocapsid protein (N-protein) in both spiked samples and clinical nasopharyngeal swab samples of COVID-19 patients within 30 min. The two working electrodes offered the limit of detection of 116 fg/mL and 150 fg/mL, respectively, with the dynamic detection range of 1-10,000 pg/mL and the sensitivity range of 2744-2936 Ω mL/pg.mm2 for the detection of N-protein. The potentiostat performed comparable or better than commercial Autolab potentiostats while it is significantly lower cost. The open-source Bi-ECDAQ presents a customizable and flexible approach towards addressing the need for rapid and accurate point-of-care electrochemical biosensors for the rapid detection of various diseases.

Electrochemical coupled immunosensing platform based on graphene oxide/gold nanocomposite for sensitive detection of Cronobacter sakazakii in powdered infant formula.

A sensitive electrochemical immunosensing platform for the detection of Cronobacter sakazakii was developed using a graphene oxide/gold (GO/Au) composite. Transmission electron microscopy showed that the Au nanoparticles, with an average size of < 30 nm, were well dispersed on the GO surface. For the detection of C. sakazakii, a polyclonal anti-C. sakazakii antibody (IgG) was covalently immobilized to the Au nanoparticles on the surface of the GO/Au composite coated glassy carbon electrode (GCE). The electrochemical sensing performance of immunofunctionalized GCE was characterized by cyclic voltammetry and differential pulse voltammetry. Under optimized conditions, in pure culture there was a linear relationship between electrical signal and C. sakazakii levels over the range 2.0 × 102-2.0 × 107 cfu/mL (R2 = 0.999), with a detection limit of 2.0 × 101 cfu/mL. The total analytical time was 15 min per sample. The C. sakazakii electrochemical immunosensing assay was able to successfully detect 2.0 × 101 cfu/mL of C. sakazakii in artificially contaminated powdered infant formula without any enrichment or pre-enrichment steps. Furthermore, the recovery rates of the C. sakazakii electrochemical immunosensing assay following spiking of powdered infant formula with different concentrations of C. sakazakii (cfu/mL) were 82.58% at 2.0 × 101 cfu/mL, 84.86% at 2.0 × 102 cfu/mL, and 95.40% at 2.0 × 103 cfu/mL. The C. sakazakii electrochemical immunosensing assay had good selectivity, reproducibility, and reactivity compared with other Cronobacter spp. and/or pathogens belonging to other genera, indicating its significant potential in the clinical diagnosis of C. sakazakii.

Functionalized nanocomposites with the optimal graphene oxide/Au ratio for amplified immunoassay of E. coli to estimate quality deterioration in dairy product.

An amplified electrochemical immunosensor was developed by exploiting poly(diallyldimethylammonium chloride)-functionalized graphene oxide and gold nanoparticles (GO-PDDA@AuNP) as sensing platform. Highlight of this work was to investigate and optimize the ratio of GO-PDDA to AuNP for sensitivity enhancement. The suitable GO-PDDA@AuNP nanocomposites were proven to not only provide an excellent biocompatible microenvironment for the immobilized antibody, but also accelerate electron transfer to enhance electrochemical signal. Moreover, the {dAb-Au-THi} nanoprobes as biorecognition elements were designed by utilizing the amplification effect of AuNPs to load detection antibody (dAb) and enormous thionine (THi), in which dAb was used for specific recognition of E. coli and THi served as electroactive species. Under optimal conditions, experimental results demonstrated that the variations (ΔI) in the peak current linearly depended on the logarithmic value of E. coli concentration from 50 to 5.0×106cfumL-1 with the detection limit of 35cfumL-1. Recovery experiments were also performed for E. coli assay in dairy product (pure fresh milk, yogurt in shelf-life and expired yogurt), and the recoveries of standard additions were in the range of 89.7-112.6%. The excellent performance of the proposed strategy indicated its promising prospect as a valuable tool to estimate the quality deterioration in dairy product.

Using silver nanoparticle and thiol graphene quantum dots nanocomposite as a substratum to load antibody for detection of hepatitis C virus core antigen: Electrochemical oxidation of riboflavin was used as redox probe.

In this study a facile green approach to employ silver nanoparticle (AgNPs) and thiol graphene quantum dots (GQD-SH) as the nanomaterial for ultrasensitive and selective detection of hepatitis C virus core antigen (HCV) have been investigated. AgNPs/GQD-SH was utilized as a substratum to load antibody for detection of hepatitis C virus core antigen. AgNPs have been immobilized on SH groups of GQDs via bonding formation of Ag-S and anti-HCV have been loaded on the electrode surface via the interaction between -NH2 group of antibody and AgNPs. Using the proposed nanocomposite provides a specific platform with increased surface which is capable of loading more antibodies to entrap the antigen. The decreasing of the electrochemical signal can be achieved after the specific recognition between antibodies and antigens. Riboflavin was used as a biological molecule with inherent properties, for the first time, as the redox probe in the development of HCV core antigen electrochemical immunosensor. Compared to the other redox probes, riboflavin is superior in its oxidization in negative potential range, where the number of interfering species for riboflavin is much fewer. The proposed immunosensor showed wide linear range from 0.05pgmL-1 to 60ngmL-1 with limit of detection of 3fgmL-1. This novel immunosensor was used to analyze the serum sample. The immunosensor provides a convenient, low-cost and simple method for HCV core antigen detection and proposes new horizons for quantitative detection of antigen in the clinical diagnosis.