Label-free Electrochemical Impedance Spectroscopy Research Articles

Deaeration of incubation buffer to minimize electrochemical impedance spectroscopy signal drift on screen-printed gold electrodes and its use for label-free detection of vancomycin

Disposable screen-printed gold electrodes (SPAuEs) are used for the development of numerous immunosensors owing to their ease of modification with biorecognition molecules. A major limitation in the development of SPAuEs based biosensors is the single-use of the electrodes leading to higher costs for sensor development, whereas multiple measurements can affect the sensor stability and reproducibility. Especially, label-free electrochemical impedance spectroscopy (EIS) based SPAuEs sensors are prone to signal drift resulting in false-positive or false-negative data questioning the reliability of the assay. We found that repeated EIS measurements on SPAuEs in ferri- and ferrocyanide solution yielded a significant increase in resistance to charge transfer (Rct) value (positive signal drift) even in a control solution (without the target analyte). The positive signal drift follows the typical pattern of a concentration-dependent calibration curve, rendering repeated measurement error-prone for sensing purposes. In this study, it is shown that the incubation of the SPAuEs in nitrogen purged deaerated phosphate buffered saline (N-PBS) minimized this signal drift in repeated/multiple EIS measurements. This method was then used to develop a peptide-based EIS biosensor for the detection of Vancomycin, a last-line antibiotic used for the treatment of severe multidrug-resistant bacterial infection. This protocol can be followed for the biosensing of other clinically important biomarkers.

Development of an Integrated Multi-Method Electrochemical Biosensor for Rapid-on-Site Detection and Quantification of Cyanotoxins

The accumulation of Cyanotoxins (CTXs) in water reservoirs is an emerging global concern. Exposure to microcystin-LR (MC-LR), among the most common CTXs, is associated with hepatotoxicity and carcinogenesis and leads to both acute and chronic damages. Stringent regulation has recently been imposed by the World Health Organization, limiting the allowed MC-LR concentration in drinking water to <1 μg/L. Current methods for MC-LR detection require expensive equipment, trained personnel, and labor-intensive preparation. Therefore, there is an urgent need for an affordable, on-site diagnostic tool providing a rapid quantitative determination of MC-LR in various water types. In this study, we have developed an electrochemical (EC) biosensor that comprised of a miniaturized and multiplexed microchips, functionalized with anti-MC-LR antibodies, and MC-LR detection is based on the transduction of bimolecular binding into an electrochemical signal. The transduction method applied is based on the comparison of two (2) electrochemical techniques: a label-free Electrochemical Impedance Spectroscopy (EIS) a labeled Electrochemical Immunoassay (ECI) detection of MC-LR in <10 min. The former, the EIS-based detection method, demonstrated the feasibility of detecting ultra-low (3 ppt or ng/L) MC-LR concentrations, and preliminary results also showed that the biochip demonstrated its use in rapid quantitative detection of microcystins in raw cyanobacterial cultures. However, the latter, ECI-based detection method showed a detection limit of 3µg/L (3ppb) of purified MC-LR concentrations, which is an order of magnitude less sensitive than the EIS-based detection method. These LODs obtained from the two methods are superior to currently employed solid-phase immunoassays and other analytical methods. Based on our findings, we anticipate that in the near future electrochemical biosensors would be an essential tool in water monitoring and environmental diagnostics. Figure 1

Label-Free Detection of CA19-9 Using a BSA/Graphene-Based Antifouling Electrochemical Immunosensor.

Evaluating the levels of the biomarker carbohydrate antigen 19-9 (CA19-9) is crucial in early cancer diagnosis and prognosis assessment. In this study, an antifouling electrochemical immunosensor was developed for the label-free detection of CA19-9, in which bovine serum albumin (BSA) and graphene were cross-linked with the aid of glutaraldehyde to form a 3D conductive porous network on the surface of an electrode. The electrochemical immunosensor was characterized through the use of transmission electron microscopy (TEM), scanning electron microscopy (SEM), atomic force microscope (AFM), UV spectroscopy, and electrochemical methods. The level of CA19-9 was determined through the use of label-free electrochemical impedance spectroscopy (EIS) measurements. The electron transfer at the interface of the electrode was well preserved in human serum samples, demonstrating that this electrochemical immunosensor has excellent antifouling performance. CA19-9 could be detected in a wide range from 13.5 U/mL to 1000 U/mL, with a detection limit of 13.5 U/mL in human serum samples. This immunosensor also exhibited good selectivity and stability. The detection results of this immunosensor were further validated and compared using an enzyme-linked immunosorbent assay (ELISA). All the results confirmed that this immunosensor has a good sensing performance in terms of CA19-9, suggesting its promising application prospects in clinical applications.

Evaluation of the passive permeability of antidepressants through pore-suspended lipid bilayer

HypothesisThe antidepressant drug imipramine, and its metabolite desipramine show different extents of interaction with, and passive permeation through, cellular membrane models, with the effects depending on the membrane composition. Through multimodal interrogation, we can observe that the drugs have a direct impact on the physicochemical properties of the membrane, that may play a role in their pharmacokinetics. ExperimentsMicrocavity pore-suspended lipid bilayers (MSLBs) of four different compositions, each with a different headgroup charge namely; zwitterionic dioleoylphosphatidylcholine (DOPC), mixed DOPC and negatively charged dioleoylphosphatidylglycerol (DOPG) (3:1), mixed DOPC and positively charged dioleoyltrimethylammoniumpropane (DOTAP) (3:1), and with increasing complex composition mimicking blood-brain-barrier (BBB) were prepared on gold and polydimethylsiloxane (PDMS) substrates using a Langmuir-Blodgett-vesicle fusion method. The molecular interaction and permeation of antidepressants, imipramine, and its metabolite desipramine with the lipid bilayers were evaluated using highly sensitive label-free electrochemical impedance spectroscopy (EIS) and surface-enhanced Raman spectroscopy (SERS). Drug-induced membrane packing/fluidity alterations were assessed using fluorescence lifetime imaging (FLIM) and fluorescence lifetime correlation spectroscopy (FLCS) of MSLB over microfluidic PDMS array. FindingsUsing EIS to evaluate in real-time membrane admittance changes, we found that imipramine greatly increases the ion permeability of negatively charged DOPC:DOPG (3:1) membranes. The effect was observed also at neutral (DOPC) and to a lesser extent at positively charged DOPC:DOTAP(3:1) membranes. In contrast, desipramine had a much weaker impact on ion permeability across all bilayer compositions. Temporal capacitance data show that desipramine intercalates at negatively charged membrane thereby increasing the thickness of the membrane. The overall kinetics of the imipramine permeation is higher than that of desipramine. This was confirmed using SERS, which also provides an evaluation of drug passive permeation based on arrival time across the membrane. Using FLCS, we found that imipramine increases the lipid membrane fluidity, whereas desipramine lowers it, with the exception of the negatively charged membrane. A translocation rate pharmacokinetics model was established for the first time at the MSLB platform by real-time monitoring of the variation in membrane resistance of pristine DOPC and blood-brain-barrier (BBB) membrane.

A diagnostic electrochemical aptasensor development for sCD80 protein detection in human serum

Elevating soluble CD80 (sCD80) in human serum is a natural response to autoimmune diseases such as rheumatoid arthritis (RA). The level of sCD80 is associated with RA development and prognosis; therefore, it is potentially used as a biomarker. sCD80 is commonly measured in human serum using immunoassays (e.g., ELISA) with multiple drawbacks, mainly cross-reactivity. Aptamer-based biosensors (aptasensors) development for quantifying and detecting different biological molecules demonstrates applicability in next-generation medicine and biomarker detection. Herein, we selected a specific aptamer for sCD80 by conventional in-vitro selection process (SELEX) with the high-affinity aptamer (Kd = 47.69 nM). A sensitive aptasensor, for the first time, was developed on a screen-printed gold electrode (AuSPE) platform compatible with easy-to-use label-free electrochemical impedance spectroscopy. The immobilization of the aptamer on the gold surface and the presence of sCD80 in a complex with the aptamer were characterized by photo-induced force microscopy, which revealed the uniform assembly of the aptamer monolayer and the distribution of sCD80 on the electrode surface. The developed aptasensor showed a linear performance (0.025–10.0 nM of protein) with a detection limit of 8.0 pM. Furthermore, the aptasensor was tested in a biological matrix, where a linear signal was observed for the increased amount of spiked sCD80 (R2 = 0.9887). The recovery of the spiked amounts ranged from 105 to 125% with coefficient of variation (CV%) <7%, which supported the applicability of this sensor in detecting sCD80 for diagnosis.

Development of label-free electrochemical impedance spectroscopy for the detection of HLA-B*15:02 and HLA-B*15:21 for the prevention of carbamazepine-induced Stevens-Johnson syndrome

BackgroundCarbamazepine (CBZ) is an FDA-approved anticonvulsant that is widely used to treat epilepsy, bipolar disorder, trigeminal neuralgia and chronic pain. Several studies have reported a strong association between HLA-B*15:02 and carbamazepine-induced Stevens–Johnson syndrome (SJS) or toxic epidermal necrolysis (TEN). However, the HLA-B75 serotype (HLA-B*15:02, HLA-B*15:08, HLA-B*15:11 and HLA-B*15:21) has been found in patients with carbamazepine-induced SJS/TEN. MethodsThis study aimed to develop label-free electrochemical impedance spectroscopy (EIS) for the detection of HLA-B*15:02 and HLA-B*15:21 after PCR-SSP amplification. A total of 208 DNA samples were tested. The impedance was measured and compared to standard gel electrophoresis. ResultsThe developed label-free EIS identified HLA-B*15:02 and HLA-B*15:21 alleles with 100% sensitivity (95% CI: 86.773%–100.000%) and 95.05% specificity (95% CI: 90.821%–97.714%), comparable to commercial DMSc 15:02 detection kits. ConclusionsWe successfully developed a novel PCR-SSP associated with signal impedance changes to detect the HLA-B*15:02 allele and HLA-B*15:21 without downstream amplicon size analysis that is suitable for screening individuals before indication of CBZ therapy.

A Label-Free Electrochemical Impedimetric Immunosensor with Biotinylated-Antibody for SARS-CoV-2 Nucleoprotein Detection in Saliva.

The timely detecting of SARS-CoV-2 coronavirus antigens for infection validation is an urgent request for COVID-19 pandemic control. This study constructed label-free electrochemical impedance spectroscopy (EIS)-based immunosensors based on gold nanostructured screen-printed carbon electrodes (AuNS/SPCEs) to detect the SARS-CoV-2 nucleocapsid protein (N-protein) in saliva. Using short-chain 3-mercaptopropionic acid (MPA) as a linker to covalently bond streptavidin (SA) and bovine serum albumin (BSA) for controlling the oriented immobilization of the biotinylated anti-N-protein antibody (BioAb) can offer a greater sensitivity, a lower limit of detection (LOD), and better reproducibility of immunosensors (defined as BioAb/SA-BSA/MPA/AuNS/SPCEs) than the antibody randomly immobilized immunosensors and the long-chain 11-mercaptoundecanoic acid (MUA)-modified immunosensors (BioAb/SA-BSA/MUA/AuNS/SPCEs). The BioAb/SA-BSA/MPA/AuNS/SPCE-based immunosensors presented good linearity from 0.01 ng/mL to 100 ng/mL and a low LOD of 6 pg/mL in a phosphate buffer solution (PBS) and PBS-diluted saliva. Moreover, the immunosensor exhibited little cross-activity with other viral antigens such as MERS-CoV N-protein, influenza A N-protein, influenza B N-protein, and SARS-CoV-2 spike protein, indicating the high specificity of the immunosensors. The disposable label-free EIS-based immunosensors have promising potential in facilitating the rapid and sensitive tests of saliva-based COVID-19 diagnostics.

Fetuin derivatised surface for evaluation of neuraminidase inhibitors of Peste des petits ruminants virus on electrochemical impedance sensor

Neuraminidase (NA) is a viral glycoprotein present in the surface of viruses belonging to paramyxoviridae and orthomyxoviridae. It helps in viral attachment and movement in the airways. It is also essential for release of viral progeny from the infected cells. Thus, anti-neuraminic agents have the potential to obstruct infection well before the virus interacts with the host. In the present study, we describe a label-free electrochemical impedance spectroscopy to characterise the NA inhibitors of plant origin against the Peste des petits ruminants virus NA. A decrease in charge transfer resistance, ΔRct (>50%;p<0.05) was observed for morin and eugenol when incubated against NA validates their anti-neuraminidase activity. Further, reduced virus propagation in vitro corroborated the efficient anti-neuraminic activity of morin and eugenol. PPRV Neuraminidase Inhibition stipulated the IC50 of 26.01 µM and 115.1 µM for morin and eugenol respectively. Additionally, we observed a selective index of both phytochemicals (16.2 for morin and 11.75 for eugenol). The proposed impedance spectroscopy method has sensitivity comparable to that of traditional, but avoiding the tedious screening system for anti- neuraminic compounds.

Determination of Vascular Endothelial Growth Factor (VEGF) in Cell Culture Medium by Gold-Coated Magnetic Nanoparticle Based Label-Free Electrochemical Impedance Spectroscopy (EIS)

A gold-coated magnetic nanoparticle (MNPs@Au) based label-free electrochemical impedance spectroscopy (EIS) sensor is reported for determining vascular endothelial growth factor (VEGF) in cell culture media. VEGF antibodies (Ab) were modified on MNPs@Au to construct an Ab-bound magnetic complex (MNPs@Au-Ab) that was magnetically immobilized on a magnetic glassy carbon electrode (mGCE). The VEGF is captured on the MNPs@Au-Ab/mGCE, leading to increased electron-transfer resistance (R et) utilized for the determination of VEGF. Combining the highly efficient immobilization of antibodies on magnetic nanocomposites with the high sensitivity of electrochemical impedance spectroscopy, an excellent dynamic range for the VEGF concentration from 1.0 × 10−2 to 1.0 × 104 ng mL−1 was obtained with a detection limit of 2.43 pg mL−1. Quantitative determination and the investigation of the secretion of VEGF by various cells were also conducted. VEGF released by cancer cells (MDA-MB-231 cells and HCT116 cells) was significantly higher than for normal cells (L02 cells), which matched well with an enzyme linked immunosorbent assay (ELISA). These results show that the secretion of VEGF is associated with the metastatic level of cells. The reported sensor is expected to be applied in studies of cancer and tumor metastasis.

An analysis method to detect the presence of DNA using electrochemical impedance spectroscopy (EIS) for real-time PCR

This paper reports a technique for detecting the presence of DNA in real-time PCR using label-free electrochemical impedance spectroscopy. The change in the real and imaginary parts of the impedance with the number of PCR cycles indicates the concentration of amplified DNA. In general, the real part of the impedance tends to increase with the number of PCR cycles over most of the measurement frequency range (100 Hz–1 MHz), whereas the imaginary part of the impedance increases in the negative direction. The optimal frequency for impedance analysis is determined by the performance index (PI), as one of the figures of merit, which quantitatively indicates how monotonously the impedance changes with the PCR cycle number. The detectability of DNA in the sample is scrutinized by analyzing the impedance at the optimal frequency (3.984 kHz and 20.02 kHz for the real and imaginary parts of the impedance, respectively) obtained from the performance index. The proposed technique can be used to quantitatively analyze PCR by applying simple electrochemical impedance spectroscopy (EIS) technology so that a label-free real-time monitoring system has high potential for a portable PCR system.

Hyaluronic acid-based label-free electrochemical impedance analysis for cancer cell quantification and CD44 expression

In this study, a novel label-free electrochemical impedance spectroscopy (EIS) sensor was developed by successfully conjugating hyaluronic acid (HA) with bovine serum albumin (BSA)-modified gold nanoparticles (GNPs) for cancer cell quantification and CD44 expression analysis. Based on the molecular recognition of HA, the CD44-overexpressing cancer cells can be captured on the nanocomposites (HA-BSA-GNPs) modified electrode, increasing the electron-transfer resistance (Ret). The change of Ret was measured by the EIS to quantify the number of cancer cells and evaluate the CD44 expression level on the cell surface. Under optimal conditions, the change of Ret was proportional to the logarithm of the cell concentration, namely from 2.0 × 102 to 3.0 × 105 cells mL−1 with a detection limit of 128 cells mL−1 for the MDA-MB-231 cells. Moreover, compared to HCT116 and L02 cells, the MDA-MB-231 cells exhibited a distinctly higher expression level of CD44. This indicates that the MDA-MB-231 cells have greater metastatic potential. The proposed sensor may find potential applications in cancer monitoring and research on tumor metastasis.

Clinically comparable impedimetric immunosensor for serum alkaline phosphatase detection based on electrochemically engineered Au-nano-Dendroids and graphene oxide nanocomposite

In this work, we demonstrate label-free electrochemical impedance spectroscopy (EIS) based alkaline phosphatase (ALP) detection using gold nanoparticles (AuNPs), electrochemically engineered Au-nano-Dendroids, and graphene oxide (GO) nanocomposite. These nanomaterials were sequentially deposited on to the screen-printed carbon electrode (SPCE) and antibodies against ALP (anti-ALP) were immobilized using carbodiimide bioconjugation process. The sensor probe has been characterized extensively using TEM, EDX, SAED, XRD, FE-SEM, FTIR, DIC, and electrochemical techniques. The analytical performance of fabricated biosensor has been evaluated using EIS, where linear dynamic range and limit of detection were obtained to be 100–1000 U/L and 9.10 (±0.12) U/L, respectively. The developed biosensor showed high selectivity towards ALP with negligible interference (ksel « 1; n = 3) due to coexisting molecules. The sensor probe has successfully recovered ALP between 108.84% and 172.50% (n = 3) in human serum samples. The sensor has been used to estimate ALP in clinical serum samples, where the level was found to be 83.15 U/L and was comparable with standard technique used in the hospitals. The shelf life, stability, and reproducibility have also been evaluated.

Toward a Label-Free Electrochemical Impedance Immunosensor Design for Quantifying Cortisol in Tears.

Cortisol is a viable biomarker for monitoring physiological, occupational, and emotional stress and is normally present in tear fluid at approximately 40 nM, or higher as a result of stress. We present characterization and quantification of cortisol via several electrochemical methods versus the standard enzyme-linked immunosorbent assay, commonly known as ELISA. We also present a prototyped design of a disposable test strip and handheld sensor based on label-free electrochemical impedance spectroscopy to quantify cortisol levels in tear fluid within approximately 90 seconds. Electrochemical characterization of the cortisol molecule was conducted using cyclic voltammetry, amperometric i-t, and square wave voltammetry. Lower limits of detection for these techniques were not sufficient to quantify cortisol and phycological tear ranges: 0.1 M, 0.23 M, and 193 M for cyclic voltammetry, amperometric i-t, and square wave voltammetry, respectively. However, electrochemical impedance spectroscopy (EIS) was to be the best mode of cortisol quantification and comparison to ELISA technique (detection range of ~ 138 pM - 552 nM). The initial EIS biosensor obtained a lower limit of detection of 59.76 nM with an approximate 10% relative standard deviation. The cortisol assay and tear collection prototype presented here offer a highly reproducible and ultra-low level of detection with a label-free and rapid response.

Hyaluronate-Functionalized Graphene for Label-Free Electrochemical Cytosensing.

Electrochemical sensors for early tumor cell detection are currently an important area of research, as this special region directly improves the efficiency of cancer treatment. Functional graphene is a promising alternative for selective recognition and capture of target cancer cells. In our work, an effective cytosensor of hyaluronate-functionalized graphene (HG) was prepared through chemical reduction of graphene oxide. The as-prepared HG nanostructures were characterized with Fourier transform infrared spectroscopy and transmission electron microscopy coupled with cyclic voltammograms and electrochemical impedance spectroscopy, respectively. The self-assembly of HG with ethylene diamine, followed by sodium hyaluronate, enabled the fabrication of a label-free electrochemical impedance spectroscopy cytosensor with high stability and biocompatibility. Finally, the proposed cytosensor exhibited satisfying electrochemical behavior and cell-capture capacity for human colorectal cancer cells HCT-116, and also displayed a wide linear range, from 5.0 × 102 cells∙mL−1 to 5.0 × 106 cells∙mL−1, and a low detection limit of 100 cells∙mL−1 (S/N = 3) for quantification. This work paves the way for graphene applications in electrochemical cytosensing and other bioassays.

Label-Free and Redox Markers-Based Electrochemical Aptasensors for Aflatoxin M1 Detection.

We performed a comparative analysis of the sensitivity of aptamer-based biosensors for detection mycotoxin aflatoxin M1 (AFM1) depending on the method of immobilization of DNA aptamers and method of the detection. Label-free electrochemical impedance spectroscopy (EIS) and differential pulse voltammetry (DPV) for ferrocene labeled neutravidin layers were used for this purpose. Amino-modified DNA aptamers have been immobilized at the surface of polyamidoamine dendrimers (PAMAM) of fourth generation (G4) or biotin-modified aptamers were immobilized at the neutravidin layer chemisorbed at gold surface. In the first case the limit of detection (LOD) has been determined as 8.47 ng/L. In the second approach the LOD was similar 8.62 ng/L, which is below of allowable limits of AFM1 in milk and milk products. The aptasensors were validated in a spiked milk samples with good recovery better than 78%. Comparative analysis of the sensitivity of immuno- and aptasensors was also performed and showed comparable sensitivity.

Impedance Spectroscopy and Electrophysiological Imaging of Cells With a High-Density CMOS Microelectrode Array System.

A monolithic multi-functional CMOS microelectrode array system was developed that enables label-free electrochemical impedance spectroscopy of cells in vitro at high spatiotemporal resolution. The electrode array includes 59,760 platinum microelectrodes, densely packed within a 4.5 mm × 2.5 mm sensing region at a pitch of 13.5 μm. A total of 32 on-chip lock-in amplifiers can be used to measure the impedance of any arbitrarily chosen subset of electrodes in the array. A sinusoidal voltage, generated by an on-chip waveform generator with a frequency range from 1 Hz to 1 MHz, was applied to the reference electrode. The sensing currents through the selected recording electrodes were amplified, demodulated, filtered, and digitized to obtain the magnitude and phase information of the respective impedances. The circuitry consumes only 412 μW at 3.3 V supply voltage and occupies only 0.1 mm2, for each channel. The system also included 2048 extracellular action-potential recording channels on the same chip. Proof of concept measurements of electrical impedance imaging and electrophysiology recording of cardiac cells and brain slices are demonstrated in this paper. Optical and impedance images showed a strong correlation.

Anti-Tau Antibodies Based Electrochemical Sensor for Detection of Tau Protein Biomarkers

Neuronal tau protein is one of the biomarkers of neurodegeneration which remains without a cure and differential diagnosis. Hence, novel methods are needed for detection of tau protein. Toward this aim, the detection of full-length tau441 protein was achieved using the electrochemical immunoassay based on direct immobilization of anti-tau antibodies on Au surface by using an active N-hydroxysuccinimide ester coupling chemistry (Ab-Au). The electrochemical impedance spectroscopy was used to monitor tau441 binding to Ab-Au in the presence [Fe(CN)6]3−/4− redox probe. The reduction in the charge-transfer resistance, Rct, was observed upon tau441 binding to Ab-Au. Detection of tau441 was observed in μM to nM range with high specificity and selectivity. The electrochemical immunosensor exhibited minimal non-specific binding to bovine serum albumin. Tau441 did not bind to Au surface functionalized with non-tau-specific antibody MOPC-21. The electrochemical biosensor was optimized by screening various anti-tau antibodies. The tau441 binding to anti-tau antibodies was also measured using Enzyme-Linked Immunosorbent Assay (ELISA). The label-free electrochemical impedance spectroscopy immunosensor may be used for selective detection of tau protein in biological fluids, as a biomarker of neurodegeneration.

Rapid and Label-Free Detection of Interferon Gamma via an Electrochemical Aptasensor Comprising a Ternary Surface Monolayer on a Gold Interdigitated Electrode Array.

A label-free electrochemical impedance spectroscopy (EIS) aptasensor for rapid detection (<35 min) of interferon-gamma (IFN-γ) was fabricated by immobilizing a RNA aptamer capture probe (ACP), selective to IFN-γ, on a gold interdigitated electrode array (Au IDE). The ACP was modified with a thiol group at the 5' terminal end and subsequently co-immobilized with 1,6-hexanedithiol (HDT) and 6-mercapto-1-hexanolphosphate (MCH) to the gold surface through thiol-gold interactions. This ACP/HDT-MCH ternary surface monolayer facilitates efficient hybridization with IFN-γ and displays high resistance to nonspecific adsorption of nontarget proteins [i.e., fetal bovine serum (FBS) and bovine serum albumin (BSA)]. The Au IDE functionalized with ACP/HDT-MCH was able to measure IFN-γ in actual FBS solution with a linear sensing range from 22.22 pM to 0.11 nM (1-5 ng/mL) and a detection limit of 11.56 pM. The ability to rapidly sense IFN-γ within this sensing range makes the developed electrochemical platform conducive toward in-field disease detection of a variety of diseases including paratuberculosis (i.e., Johne's Disease). Furthermore, experimental results were numerically validated with an equivalent circuit model that elucidated the effects of the sensing process and the influence of the immobilized ternary monolayer on signal output. This is the first time that ternary surface monolayers have been used to selectively capture/detect IFN-γ on Au IDEs.

Label-free electrochemical impedance peptide-based biosensor for the detection of cardiac troponin I incorporating gold nanoparticles modified carbon electrode

A label-free electrochemical impedance spectroscopy (EIS) biosensor for highly sensitive detection of cardiac troponin I (TnI) as a model target was developed by employing gold nanoparticle (GNPs) modified glassy carbon electrode (GCE) as a base electrode. A special peptide (CFYSHSFHENWPS) was used as molecular recognition probe. GNPs were firstly electrochemically deposited onto the surface of GCE and polyethylene glycol was then used to block any remaining sites of the surface of GCE. The biosensor was fabricated by thiol self-assembling the peptide probe onto the surface of gold nanoparticles and finally blocking any remaining sites of the surface of GNPs using 6-mercapto-1-hexanol. A suitable equivalent electrical circuit consisting of parallel double Randles circuit with phase angle elements greatly fits the experiment electrochemical cell. Upon the binding of TnI, the charge transfer resistance (Rct) of the biosensor was logarithmically direct proportional to the concentration of TnI in the range from 15.5pg/mL to 1.55ng/mL with a detection limit of 3.4pg/mL. It was found that GNPs modified GCE as a platform for the immobilization of the peptide can greatly reduce the background EIS signal and enhance the EIS response to the target. This biosensor has been used for assay of real serum sample with satisfactory results. This work demonstrates that GNPs modified GCE for the immobilization of molecular recognition element is promising platform for the development of label-free electrochemical biosensors for the determination of special proteins.

A novel label-free and reusable electrochemical cytosensor for highly sensitive detection and specific collection of CTCs

Circulating tumor cells (CTCs) contain a great deal of information of tumor phenotype. Therefore, highly sensitive detection and specific enrichment of CTCs are of intense interest. Herein, a label-free electrochemical impedance spectroscopy cytosensor with effective surface recognition between specific epithelial cell adhesion molecules (EpCAM) over-expressed on the cell membrane and EpCAM aptamer was developed for the detection of CTCs. After immobilization of 6-mercapto-1-hexanol (MCH) onto the gold electrode, the capture probe can be directionally inserted in MCH interspaces, which can improve the sensitivity of the cytosensor. A wide detection range from 30 to 1×106cellsmL−1 with a detection limit as low as 10cellsmL−1 is reached on the condition of acceptable stability and reproducibility. The cytosensor can easily distinguish CTCs from the real blood sample due to the specific combination of EpCAM and EpCAM aptamer. Furthermore, the cytosensor can be reused 8 times and enrich CTCs by Uracil DNA Excision Mix specific cleaving the deoxyuridines (dUs) of the aptamer. The collected CTCs can contribute to further study. Thus, we reported that this cytosensor is a promising technique for the early monitoring and therapy of cancer.