Label free, electric field mediated ultrasensitive electrochemical point-of-care device for CEA detection

AbstractDeveloping point-of-care (PoC) diagnostic platforms for carcinoembryonic antigen detection is essential. However, thefew implementations of transferring the signal amplification strategies in electrochemical sensing on paper-based platforms are not satisfactory in terms of detection limit (LOD). In the quest for pushing down LOD, majority of the research has been targeted towards development of improved nanostructured substrates for entrapping more analyte molecules and augmenting the electron transfer rate to the working electrode. But, such approaches have reached saturation. This paper focuses on enhancing the mass transport of the analyte towards the sensor surface through the application of an electric field, in graphene-ZnO nanorods heterostructure. These hybrid nanostructures have been deposited on flexible polyethylene terephthalate substrates with screen printed electrodes for PoC application. The ZnO nanorods have been functionalized with aptamers and the working sensor has been integrated with smartphone interfaced indigenously developed low cost potentiostat. The performance of the system, requiring only 50 µl analyte has been evaluated using electrochemical impedance spectroscopy and validated against commercially available ELISA kit. Limit of detection of 1 fg/ml in human serum with 6.5% coefficient of variation has been demonstrated, which is more than three orders of magnitude lower than the existing attempts on PoC device.

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Demonstration of a Label-Free and Low-Cost Optical Cavity-Based Biosensor Using Streptavidin and C-Reactive Protein

Biosensors ◽

10.3390/bios11010004 ◽

2020 ◽

Vol 11 (1) ◽

pp. 4

Author(s):

Donggee Rho ◽

Seunghyun Kim

Keyword(s):

Limit Of Detection ◽

Point Of Care ◽

Low Cost ◽

Optical Cavity ◽

Sample Volume ◽

Small Sample ◽

Label Free ◽

C Reactive Protein ◽

Reactive Protein ◽

Cavity Structure

An optical cavity-based biosensor (OCB) has been developed for point-of-care (POC) applications. This label-free biosensor employs low-cost components and simple fabrication processes to lower the overall cost while achieving high sensitivity using a differential detection method. To experimentally demonstrate its limit of detection (LOD), we conducted biosensing experiments with streptavidin and C-reactive protein (CRP). The optical cavity structure was optimized further for better sensitivity and easier fluid control. We utilized the polymer swelling property to fine-tune the optical cavity width, which significantly improved the success rate to produce measurable samples. Four different concentrations of streptavidin were tested in triplicate, and the LOD of the OCB was determined to be 1.35 nM. The OCB also successfully detected three different concentrations of human CRP using biotinylated CRP antibody. The LOD for CRP detection was 377 pM. All measurements were done using a small sample volume of 15 µL within 30 min. By reducing the sensing area, improving the functionalization and passivation processes, and increasing the sample volume, the LOD of the OCB are estimated to be reduced further to the femto-molar range. Overall, the demonstrated capability of the OCB in the present work shows great potential to be used as a promising POC biosensor.

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Glycoprofiling of cancer biomarkers: Label-free electrochemical lectin-based biosensors

Open Chemistry ◽

10.1515/chem-2015-0082 ◽

2015 ◽

Vol 13 (1) ◽

Cited By ~ 31

Author(s):

Dominika Pihíková ◽

Peter Kasák ◽

Jan Tkac

Keyword(s):

Electrochemical Impedance ◽

Early Stage ◽

Point Of Care ◽

Low Cost ◽

Warning Signal ◽

Cancer Biomarkers ◽

Cancer Diagnostics ◽

Label Free ◽

Glycosylation Pattern ◽

Human Proteins

AbstractGlycosylation of biomolecules is one of the most prevalent post- and co-translational modification in a human body, with more than half of all human proteins being glycosylated. Malignant transformation of cells influences glycosylation machinery resulting in subtle changes of the glycosylation pattern within the cell populations as a result of cancer. Thus, an altered terminal glycan motif on glycoproteins could provide a warning signal about disease development and progression and could be applied as a reliable biomarker in cancer diagnostics. Among all highly effective glycoprofiling tools, label-free electrochemical impedance spectroscopy (EIS)-based biosensors have emerged as especially suitable tool for point-of-care early-stage cancer detection. Herein, we highlight the current challenges in glycoprofiling of various cancer biomarkers by ultrasensitive impedimetric-based biosensors with low sample consumption, low cost fabrication and simple miniaturization. Additionally, this review provides a short introduction to the field of glycomics and lectinomics and gives a brief overview of glycan alterations in different types of cancer.

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Companion and Point-of-Care Sensor System for Rapid Multiplexed Detection of a Panel of Infectious Disease Markers

SLAS TECHNOLOGY Translating Life Sciences Innovation ◽

10.1177/2472630317696779 ◽

2017 ◽

pp. 247263031769677

Author(s):

Anjan Panneer Selvam ◽

Shalini Prasad

Keyword(s):

Whole Blood ◽

Dynamic Range ◽

Electrochemical Impedance ◽

Limit Of Detection ◽

Point Of Care ◽

Lipoteichoic Acid ◽

Impedance Change ◽

Label Free ◽

Nylon Membrane ◽

Multiplexed Detection

A nanochannel-based electrochemical biosensor has been demonstrated for rapid and multiplexed detection of a panel of three biomarkers associated with rapid detection of sepsis. The label-free biosensor detected procalcitonin (PCT), lipoteichoic acid (LTA), and lipopolysaccharide (LPS) from human whole blood. The biosensor comprises a nanoporous nylon membrane integrated onto a microelectrode sensor platform for nanoconfinement effects. Charge perturbations due to biomarker binding are recorded as impedance changes using electrochemical impedance spectroscopy. The measured impedance change is used to quantitatively determine the concentration of the three biomarkers using antibody receptors from the tested sample. We were successful in detecting and quantifying the three biomarkers from whole blood. The limit of detection was 0.1 ng/mL for PCT and 1 µg/mL for LPS and LTA. The sensor was able to demonstrate a dynamic range of detection from 01.1 ng/mL to 10 µg/mL for PCT and from 1 µg/mL to 1000 µg/mL for LPS and LTA biomarkers. This novel technology has promising preliminary results toward the design of sensors for rapid and sensitive detection of the three panel biomarkers in whole blood toward diagnosis and classification of sepsis.

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Ultra-low HIV-1 p24 detection limits with a bioelectronic sensor

Analytical and Bioanalytical Chemistry ◽

10.1007/s00216-019-02319-7 ◽

2019 ◽

Vol 412 (4) ◽

pp. 811-818 ◽

Cited By ~ 10

Author(s):

Eleonora Macchia ◽

Lucia Sarcina ◽

Rosaria Anna Picca ◽

Kyriaki Manoli ◽

Cinzia Di Franco ◽

...

Keyword(s):

Capsid Protein ◽

Single Molecule ◽

Limit Of Detection ◽

Point Of Care ◽

Low Cost ◽

Wide Field ◽

Label Free ◽

Limit Of Quantification ◽

Great Relevance ◽

Hiv 1

AbstractEarly diagnosis of the infection caused by human immunodeficiency virus type-1 (HIV-1) is vital to achieve efficient therapeutic treatment and limit the disease spreading when the viremia is at its highest level. To this end, a point-of-care HIV-1 detection carried out with label-free, low-cost, and ultra-sensitive screening technologies would be of great relevance. Herein, a label-free single molecule detection of HIV-1 p24 capsid protein with a large (wide-field) single-molecule transistor (SiMoT) sensor is proposed. The system is based on an electrolyte-gated field-effect transistor whose gate is bio-functionalized with the antibody against the HIV-1 p24 capsid protein. The device exhibits a limit of detection of a single protein and a limit of quantification in the 10 molecule range. This study paves the way for a low-cost technology that can quantify, with single-molecule precision, the transition of a biological organism from being “healthy” to being “diseased” by tracking a target biomarker. This can open to the possibility of performing the earliest possible diagnosis.

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Homocysteine Solution-Induced Response in Aerosol Jet Printed OECTs by Means of Gold and Platinum Gate Electrodes

International Journal of Molecular Sciences ◽

10.3390/ijms222111507 ◽

2021 ◽

Vol 22 (21) ◽

pp. 11507

Author(s):

Pasquale D’Angelo ◽

Mario Barra ◽

Patrizia Lombari ◽

Annapaola Coppola ◽

Davide Vurro ◽

...

Keyword(s):

High Performance ◽

Electrochemical Impedance ◽

Limit Of Detection ◽

Biological Fluids ◽

Point Of Care ◽

Low Cost ◽

Induced Response ◽

Gate Electrodes ◽

Standard Procedures ◽

Experimental Protocols

Homocysteine (Hcy) is a non-protein, sulfur-containing amino acid, which is recognized as a possible risk factor for coronary artery and other pathologies when its levels in the blood exceed the normal range of between 5 and 12 μmol/L (hyperhomocysteinemia). At present, standard procedures in laboratory medicine, such as high-performance liquid chromatography (HPLC), are commonly employed for the quantitation of total Hcy (tHcy), i.e., the sum of the protein-bound (oxidized) and free (homocystine plus reduced Hcy) forms, in biological fluids (particularly, serum or plasma). Here, the response of Aerosol Jet-printed organic electrochemical transistors (OECTs), in the presence of either reduced (free) and oxidized Hcy-based solutions, was analyzed. Two different experimental protocols were followed to this end: the former consisting of gold (Au) electrodes’ biothiol-induced thiolation, while the latter simply used bare platinum (Pt) electrodes. Electrochemical impedance spectroscopy (EIS) analysis was performed both to validate the gold thiolation protocol and to gain insights into the reduced Hcy sensing mechanism by the Au-gated OECTs, which provided a final limit of detection (LoD) of 80 nM. For the OECT response based on Platinum gate electrodes, on the other hand, a LoD of 180 nM was found in the presence of albumin-bound Hcy, with this being the most abundant oxidized Hcy-form (i.e., the protein-bound form) in physiological fluids. Despite the lack of any biochemical functionalization supporting the response selectivity, the findings discussed in this work highlight the potential role of OECT in the development of low-cost point-of-care (POC) electronic platforms that are suitable for the evaluation, in humans, of Hcy levels within the physiological range and in cases of hyperhomocysteinemia.

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A Graphene-PEDOT:PSS Modified Paper-Based Aptasensor for Electrochemical Impedance Spectroscopy Detection of Tumor Marker

Sensors ◽

10.3390/s20051372 ◽

2020 ◽

Vol 20 (5) ◽

pp. 1372 ◽

Cited By ~ 4

Author(s):

Yi-Kuang Yen ◽

Chen-Hsiang Chao ◽

Ya-Shin Yeh

Keyword(s):

Electrochemical Impedance Spectroscopy ◽

Impedance Spectroscopy ◽

Electrochemical Impedance ◽

Limit Of Detection ◽

Point Of Care ◽

Low Cost ◽

Serum Samples ◽

Analytical Instruments ◽

Buffer Solutions ◽

Early Cancer Diagnosis

A graphene and poly (3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS) modified conductive paper-based electrochemical impedance spectroscopy (EIS) aptasensor has been successfully fabricated by a simple and continuous coating process. A graphene/PEDOT:PSS modified paper electrode forms the nanocomposite providing a conductive and sensitive substrate for further aptamer functionalization of the biosensor. This low-cost paper-based aptasensor exhibits its sensitivity to carcinoembryonic antigens (CEA) in standard buffer solutions and human serum samples in a linear range of 0.77–14 ng·mL−1. The limit of detection (LOD) is found to be 0.45 ng·mL−1 and 1.06 ng·mL−1 for CEA in both samples, separately. This aptamer-based sensing device was also evaluated and received a good correlation with the immunoassay detection method. The proposed paper-based aptasensor has demonstrated its potential as a rapid simple point-of-care analytical platform for early cancer diagnosis in less developed areas where manufacturing facilities, analytical instruments, and trained specialists are limited.

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Companion and Point-of-Care Sensor System for Rapid Multiplexed Detection of a Panel of Infectious Disease Markers

SLAS TECHNOLOGY Translating Life Sciences Innovation ◽

10.1177/2211068217696779 ◽

2017 ◽

Vol 22 (3) ◽

pp. 338-347 ◽

Cited By ~ 1

Author(s):

Anjan Panneer Selvam ◽

Shalini Prasad

Keyword(s):

Whole Blood ◽

Dynamic Range ◽

Electrochemical Impedance ◽

Limit Of Detection ◽

Point Of Care ◽

Lipoteichoic Acid ◽

Impedance Change ◽

Label Free ◽

Nylon Membrane ◽

Multiplexed Detection

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Rapid and Sensitive Detection of miRNA Based on AC Electrokinetic Capacitive Sensing for Point-of-Care Applications

Sensors ◽

10.3390/s21123985 ◽

2021 ◽

Vol 21 (12) ◽

pp. 3985

Author(s):

Nan Wan ◽

Yu Jiang ◽

Jiamei Huang ◽

Rania Oueslati ◽

Shigetoshi Eda ◽

...

Keyword(s):

Limit Of Detection ◽

Point Of Care ◽

Low Cost ◽

Clinical Diagnostics ◽

Detection Methods ◽

Capacitive Sensing ◽

Application Potential ◽

Mirna Detection ◽

Mirna 25 ◽

Low Sensitivity

A sensitive and efficient method for microRNAs (miRNAs) detection is strongly desired by clinicians and, in recent years, the search for such a method has drawn much attention. There has been significant interest in using miRNA as biomarkers for multiple diseases and conditions in clinical diagnostics. Presently, most miRNA detection methods suffer from drawbacks, e.g., low sensitivity, long assay time, expensive equipment, trained personnel, or unsuitability for point-of-care. New methodologies are needed to overcome these limitations to allow rapid, sensitive, low-cost, easy-to-use, and portable methods for miRNA detection at the point of care. In this work, to overcome these shortcomings, we integrated capacitive sensing and alternating current electrokinetic effects to detect specific miRNA-16b molecules, as a model, with the limit of detection reaching 1.0 femto molar (fM) levels. The specificity of the sensor was verified by testing miRNA-25, which has the same length as miRNA-16b. The sensor we developed demonstrated significant improvements in sensitivity, response time and cost over other miRNA detection methods, and has application potential at point-of-care.

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Label-Free Electrochemical Sensor for Rapid Bacterial Pathogen Detection Using Vancomycin-Modified Highly Branched Polymers

Sensors ◽

10.3390/s21051872 ◽

2021 ◽

Vol 21 (5) ◽

pp. 1872

Author(s):

Holger Schulze ◽

Harry Wilson ◽

Ines Cara ◽

Steven Carter ◽

Edward N. Dyson ◽

...

Keyword(s):

Pathogen Detection ◽

Electrochemical Impedance ◽

Point Of Care ◽

Branched Polymers ◽

Label Free ◽

Conformation Change ◽

Gram Positive ◽

Gram Positive Bacteria ◽

Staphylococcus Carnosus ◽

Label Free Detection

Rapid point of care tests for bacterial infection diagnosis are of great importance to reduce the misuse of antibiotics and burden of antimicrobial resistance. Here, we have successfully combined a new class of non-biological binder molecules with electrochemical impedance spectroscopy (EIS)-based sensor detection for direct, label-free detection of Gram-positive bacteria making use of the specific coil-to-globule conformation change of the vancomycin-modified highly branched polymers immobilized on the surface of gold screen-printed electrodes upon binding to Gram-positive bacteria. Staphylococcus carnosus was detected after just 20 min incubation of the sample solution with the polymer-functionalized electrodes. The polymer conformation change was quantified with two simple 1 min EIS tests before and after incubation with the sample. Tests revealed a concentration dependent signal change within an OD600 range of Staphylococcus carnosus from 0.002 to 0.1 and a clear discrimination between Gram-positive Staphylococcus carnosus and Gram-negative Escherichia coli bacteria. This exhibits a clear advancement in terms of simplified test complexity compared to existing bacteria detection tests. In addition, the polymer-functionalized electrodes showed good storage and operational stability.

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Integrating high-performing electrochemical transducers in lateral flow assay

Analytical and Bioanalytical Chemistry ◽

10.1007/s00216-021-03301-y ◽

2021 ◽

Author(s):

Antonia Perju ◽

Nongnoot Wongkaew

Keyword(s):

High Performance ◽

Point Of Care ◽

Low Cost ◽

High Sensitivity ◽

Lateral Flow ◽

Analytical Performance ◽

Label Free ◽

High Performing ◽

Lateral Flow Assays ◽

Electrochemical Transducers

AbstractLateral flow assays (LFAs) are the best-performing and best-known point-of-care tests worldwide. Over the last decade, they have experienced an increasing interest by researchers towards improving their analytical performance while maintaining their robust assay platform. Commercially, visual and optical detection strategies dominate, but it is especially the research on integrating electrochemical (EC) approaches that may have a chance to significantly improve an LFA’s performance that is needed in order to detect analytes reliably at lower concentrations than currently possible. In fact, EC-LFAs offer advantages in terms of quantitative determination, low-cost, high sensitivity, and even simple, label-free strategies. Here, the various configurations of EC-LFAs published are summarized and critically evaluated. In short, most of them rely on applying conventional transducers, e.g., screen-printed electrode, to ensure reliability of the assay, and additional advances are afforded by the beneficial features of nanomaterials. It is predicted that these will be further implemented in EC-LFAs as high-performance transducers. Considering the low cost of point-of-care devices, it becomes even more important to also identify strategies that efficiently integrate nanomaterials into EC-LFAs in a high-throughput manner while maintaining their favorable analytical performance.

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