scholarly journals A Reliable BioFET Immunosensor for Detection of p53 Tumour Suppressor in Physiological-Like Environment

Sensors ◽  
2020 ◽  
Vol 20 (21) ◽  
pp. 6364
Author(s):  
Chiara Baldacchini ◽  
Antonino Francesco Montanarella ◽  
Luca Francioso ◽  
Maria Assunta Signore ◽  
Salvatore Cannistraro ◽  
...  
Keyword(s):  
Limit Of Detection ◽  
Free Field ◽  
Direct Interaction ◽  
Tumour Suppressor ◽  
Label Free ◽  
Buffer Solution ◽  
Voltage Versus ◽  
Diagnosis And Prognosis ◽  
Early Cancer Diagnosis ◽  
Permeable Polymer

The concentration of wild-type tumour suppressor p53wt in cells and blood has a clinical significance for early diagnosis of some types of cancer. We developed a disposable, label-free, field-effect transistor-based immunosensor (BioFET), able to detect p53wt in physiological buffer solutions, over a wide concentration range. Microfabricated, high-purity gold electrodes were used as single-use extended gates (EG), which avoid direct interaction between the transistor gate and the biological solution. Debye screening, which normally hampers target charge effect on the FET gate potential and, consequently, on the registered FET drain-source current, at physiological ionic strength, was overcome by incorporating a biomolecule-permeable polymer layer on the EG electrode surface. Determination of an unknown p53wt concentration was obtained by calibrating the variation of the FET threshold voltage versus the target molecule concentration in buffer solution, with a sensitivity of 1.5 ± 0.2 mV/decade. The BioFET specificity was assessed by control experiments with proteins that may unspecifically bind at the EG surface, while 100pM p53wt concentration was established as limit of detection. This work paves the way for fast and highly sensitive tools for p53wt detection in physiological fluids, which deserve much interest in early cancer diagnosis and prognosis.

scholarly journals Pre-concentration of microRNAs by LNA-modified magnetic beads for enhancement of electrochemical detection

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Serife Ustuner ◽  
Mark A. Lindsay ◽  
Pedro Estrela
Keyword(s):  
Electrochemical Detection ◽  
Peptide Nucleic Acid ◽  
Magnetic Beads ◽  
Electrochemical Impedance ◽  
Limit Of Detection ◽  
Cancer Disease ◽  
Circulating Micrornas ◽  
Diagnosis And Prognosis ◽  
Early Cancer Diagnosis ◽  
Microrna Sequence

AbstractMicroRNAs are extremely promising candidates for early cancer diagnosis and prognosis. The levels of circulating microRNAs provide valuable information about cancer disease at its early stages. However, the levels of microRNAs that need to be detected are extremely low and difficult to discriminate from a large pool of oligonucleotides. There is the need for accurate, rapid and sensitive detection methodologies for detection of microRNAs. We developed electrochemical impedance spectroscopy peptide nucleic acid (PNA)-based sensors that can detect miRNAs in diluted serum with a limit of detection of 0.38 fM. In order to further improve the accuracy and reliability of the sensors, we developed an assay using magnetic beads for simple and rapid fishing of target microRNAs from solution and its pre-concentration prior to electrochemical detection. Our methodology utilizes magnetic beads for the capture of the target microRNA from solution and brings the concentrated sample to the sensor surface. We modify the magnetic beads with locked nucleic acids (LNA), which have high affinity and specificity to their complementary microRNA sequence. The separated and concentrated microRNA is then detected using the PNA-based sensors. By exposing the sensing electrodes only to the captured microRNAs, interferences from other nucleotides or biomolecules from the sample are eliminated.

scholarly journals "Turn on" and label-free core−shell Ag@SiO2 nanoparticles-based metal-enhanced fluorescent (MEF) aptasensor for Hg2+

2015 ◽  
Vol 5 (1) ◽  
Author(s):  
Yuanfeng Pang ◽  
Zhen Rong ◽  
Rui Xiao ◽  
Shengqi Wang
Keyword(s):  
Limit Of Detection ◽  
Fluorescence Emission ◽  
High Sensitivity ◽  
Sio2 Nanoparticles ◽  
Core Shell ◽  
Label Free ◽  
Buffer Solution ◽  
Fluorescent Reporter ◽  
Turn On ◽  
T Complex

Abstract A turn on and label-free fluorescent apasensor for Hg2+ with high sensitivity and selectivity has been demonstrated in this report. Firstly, core−shell Ag@SiO2 nanoparticles (NPs) were synthetized as a Metal-Enhanced Fluorescent (MEF) substrate, T-rich DNA aptamers were immobilized on the surface of Ag@SiO2 NPs and thiazole orange (TO) was selected as fluorescent reporter. After Hg2+ was added to the aptamer-Ag@SiO2 NPs and TO mixture buffer solution, the aptamer strand can bind Hg2+ to form T-Hg2+-T complex with a hairpin structure which TO can insert into. When clamped by the nucleic acid bases, the fluorescence quanta yield of TO will be increased under laser excitation and emitted a fluorescence emission. Furthermore, the fluorescence emission can be amplified largely by the MEF effect of the Ag@SiO2 NPs. The whole experiment can be finished within 30 min and the limit of detection is 0.33 nM even with interference by high concentrations of other metal ions. Finally, the sensor was applied for detecting Hg2+ in different real water samples with satisfying recoveries over 94%.

scholarly journals Single-step label-free nanowell immunoassay accurately quantifies serum stress hormones within minutes

2021 ◽  
Vol 7 (27) ◽  
pp. eabf4401
Author(s):  
S. Reza Mahmoodi ◽  
Pengfei Xie ◽  
Daniel P. Zachs ◽  
Erik J. Peterson ◽  
Rachel S. Graham ◽  
...  
Keyword(s):  
Human Serum ◽  
Real Time ◽  
Limit Of Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Label Free ◽  
Serum Samples ◽  
Electrode Gap ◽  
Buffer Solution ◽  
Human Serum Samples ◽  
Free Cortisol

A non-faradaic label-free cortisol sensing platform is presented using a nanowell array design, in which the two probe electrodes are integrated within the nanowell structure. Rapid and low volume (≤5 μl) sensing was realized through functionalizing nanoscale volume wells with antibodies and monitoring the real-time binding events. A 28-well plate biochip was built on a glass substrate by sequential deposition, patterning, and etching steps to create a stack nanowell array sensor with an electrode gap of 40 nm. Sensor response for cortisol concentrations between 1 and 15 μg/dl in buffer solution was recorded, and a limit of detection of 0.5 μg/dl was achieved. Last, 65 human serum samples were collected to compare the response from human serum samples with results from the standard enzyme-linked immunosorbent assay (ELISA). These results confirm that nanowell array sensors could be a promising platform for point-of-care testing, where real-time, laboratory-quality diagnostic results are essential.

A microfluidic device with integrated impedance detection for λ-DNA

2003 ◽  
Vol 773 ◽  
Author(s):  
Myung-Il Park ◽  
Jonging Hong ◽  
Dae Sung Yoon ◽  
Chong-Ook Park ◽  
Geunbae Im
Keyword(s):  
Microfluidic Device ◽  
Electrochemical Impedance ◽  
Direct Detection ◽  
Full Potential ◽  
Label Free ◽  
Buffer Solution ◽  
Detection Systems ◽  
Significant Difference ◽  
Detection Of Biomolecules ◽  
Impedance Magnitude

AbstractThe large optical detection systems that are typically utilized at present may not be able to reach their full potential as portable analysis tools. Accurate, early, and fast diagnosis for many diseases requires the direct detection of biomolecules such as DNA, proteins, and cells. In this research, a glass microchip with integrated microelectrodes has been fabricated, and the performance of electrochemical impedance detection was investigated for the biomolecules. We have used label-free λ-DNA as a sample biomolecule. By changing the distance between microelectrodes, the significant difference between DW and the TE buffer solution is obtained from the impedance-frequency measurements. In addition, the comparison for the impedance magnitude of DW, the TE buffer, and λ-DNA at the same distance was analyzed.

scholarly journals Simultaneous electrochemical sensing of dihydroxy benzene isomers at cost-effective allura red polymeric film modified glassy carbon electrode

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Pattan-Siddappa Ganesh ◽  
Ganesh Shimoga ◽  
Seok-Han Lee ◽  
Sang-Youn Kim ◽  
Eno E. Ebenso
Keyword(s):  
Glassy Carbon Electrode ◽  
Glassy Carbon ◽  
Limit Of Detection ◽  
Tap Water ◽  
Differential Pulse ◽  
Oxidation Potential ◽  
Electrochemical Sensing ◽  
Carbon Electrode ◽  
Buffer Solution ◽  
Allura Red

Abstract Background A simple and simultaneous electrochemical sensing platform was fabricated by electropolymerization of allura red on glassy carbon electrode (GCE) for the interference-free detection of dihydroxy benzene isomers. Methods The modified working electrode was characterized by electrochemical and field emission scanning electron microscopy methods. The modified electrode showed excellent electrocatalytic activity for the electrooxidation of catechol (CC) and hydroquinone (HQ) at physiological pH of 7.4 by cyclic voltammetric (CV) and differential pulse voltammetric (DPV) techniques. Results The effective split in the overlapped oxidation signal of CC and HQ was achieved in a binary mixture with peak to peak separation of 0.102 V and 0.103 V by CV and DPV techniques. The electrode kinetics was found to be adsorption-controlled. The oxidation potential directly depends on the pH of the buffer solution, and it witnessed the transfer of equal number of protons and electrons in the redox phenomenon. Conclusions The limit of detection (LOD) for CC and HQ was calculated to be 0.126 μM and 0.132 μM in the linear range of 0 to 80.0 μM and 0 to 110.0 μM, respectively, by ultra-sensitive DPV technique. The practical applicability of the proposed sensor was evaluated for tap water sample analysis, and good recovery rates were observed. Graphical abstract Electrocatalytic interaction of ALR/GCE with dihydroxy benzene isomers.

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

Biosensors ◽  
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.

scholarly journals Electro-Polymerized Titan Yellow Modified Carbon Paste Electrode for the Analysis of Curcumin

Surfaces ◽  
2021 ◽  
Vol 4 (3) ◽  
pp. 191-204
Author(s):  
Edwin S. D’Souza ◽  
Jamballi G. Manjunatha ◽  
Chenthattil Raril ◽  
Girish Tigari ◽  
Huligerepura J. Arpitha ◽  
...  
Keyword(s):  
Carbon Paste Electrode ◽  
Limit Of Detection ◽  
Phosphate Buffer Solution ◽  
Modified Carbon Paste Electrode ◽  
Carbon Paste ◽  
Buffer Solution ◽  
Solution Ph ◽  
Real Sample Analysis ◽  
Titan Yellow ◽  
Paste Electrode

A modest, efficient, and sensitive chemically modified electrode was fabricated for sensing curcumin (CRC) through an electrochemically polymerized titan yellow (TY) modified carbon paste electrode (PTYMCPE) in phosphate buffer solution (pH 7.0). Cyclic voltammetry (CV) linear sweep voltammetry (LSV) and differential pulse voltammetry (DPV) approaches were used for CRC detection. PTYMCPE interaction with CRC suggests that the electrode exhibits admirable electrochemical response as compared to bare carbon paste electrode (BCPE). Under the optimized circumstances, a linear response of the electrode was observed for CRC in the concentration range 2 × 10−6 M to 10 × 10−6 M with a limit of detection (LOD) of 10.94 × 10−7 M. Moreover, the effort explains that the PTYMCPE electrode has a hopeful approach for the electrochemical resolution of biologically significant compounds. Additionally, the proposed electrode has demonstrated many advantages such as easy preparation, elevated sensitivity, stability, and enhanced catalytic activity, and can be successfully applied in real sample analysis.

scholarly journals Circulating Tumor Cells from Enumeration to Analysis: Current Challenges and Future Opportunities

Cancers ◽  
2021 ◽  
Vol 13 (11) ◽  
pp. 2723
Author(s):  
Yu-Ping Yang ◽  
Teresa M. Giret ◽  
Richard J. Cote
Keyword(s):  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Ex Vivo ◽  
Detection Sensitivity ◽  
Clinical Utilization ◽  
Diagnosis And Prognosis ◽  
Early Cancer Diagnosis ◽  
Potential Marker ◽  
Potential Applications ◽  
Downstream Analysis

Circulating tumor cells (CTCs) have been recognized as a major contributor to distant metastasis. Their unique role as metastatic seeds renders them a potential marker in the circulation for early cancer diagnosis and prognosis as well as monitoring of therapeutic response. In the past decade, researchers mainly focused on the development of isolation techniques for improving the recovery rate and purity of CTCs. These developed techniques have significantly increased the detection sensitivity and enumeration accuracy of CTCs. Currently, significant efforts have been made toward comprehensive molecular characterization, ex vivo expansion of CTCs, and understanding the interactions between CTCs and their associated cells (e.g., immune cells and stromal cells) in the circulation. In this review, we briefly summarize existing CTC isolation technologies and specifically focus on advances in downstream analysis of CTCs and their potential applications in precision medicine. We also discuss the current challenges and future opportunities in their clinical utilization.

scholarly journals Surface Plasmon Resonance Assay for Label-Free and Selective Detection of HIV-1 p24 Protein

Biosensors ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 180
Author(s):  
Lucia Sarcina ◽  
Giuseppe Felice Mangiatordi ◽  
Fabrizio Torricelli ◽  
Paolo Bollella ◽  
Zahra Gounani ◽  
...  
Keyword(s):  
Limit Of Detection ◽  
Covalent Binding ◽  
Hiv Infections ◽  
Selectivity Ratio ◽  
Selective Detection ◽  
Label Free ◽  
Self Assembled Monolayer ◽  
Label Free Detection ◽  
P24 Protein ◽  
Hiv 1

The early detection of the human immunodeficiency virus (HIV) is of paramount importance to achieve efficient therapeutic treatment and limit the disease spreading. In this perspective, the assessment of biosensing assay for the HIV-1 p24 capsid protein plays a pivotal role in the timely and selective detection of HIV infections. In this study, multi-parameter-SPR has been used to develop a reliable and label-free detection method for HIV-1 p24 protein. Remarkably, both physical and chemical immobilization of mouse monoclonal antibodies against HIV-1 p24 on the SPR gold detecting surface have been characterized for the first time. The two immobilization techniques returned a capturing antibody surface coverage as high as (7.5 ± 0.3) × 1011 molecule/cm2 and (2.4 ± 0.6) × 1011 molecule/cm2, respectively. However, the covalent binding of the capturing antibodies through a mixed self-assembled monolayer (SAM) of alkanethiols led to a doubling of the p24 binding signal. Moreover, from the modeling of the dose-response curve, an equilibrium dissociation constant KD of 5.30 × 10−9 M was computed for the assay performed on the SAM modified surface compared to a much larger KD of 7.46 × 10−5 M extracted for the physisorbed antibodies. The chemically modified system was also characterized in terms of sensitivity and selectivity, reaching a limit of detection of (4.1 ± 0.5) nM and an unprecedented selectivity ratio of 0.02.

scholarly journals Label-Free Electrochemical Biosensor Based on Au@MoS₂–PANI for Escherichia coli Detection

Chemosensors ◽  
2021 ◽  
Vol 9 (3) ◽  
pp. 49
Author(s):  
Pushap Raj ◽  
Man Hwan Oh ◽  
Kyudong Han ◽  
Tae Yoon Lee
Keyword(s):  
Escherichia Coli ◽  
Bacterial Infections ◽  
Limit Of Detection ◽  
Bacterial Pathogens ◽  
Cost Effective ◽  
Covalent Immobilization ◽  
Label Free ◽  
Self Assembled Monolayer ◽  
Detection Range ◽  
Linear Detection

Bacterial infections have become a significant challenge in terms of public health, the food industry, and the environment. Therefore, it is necessary to address these challenges by developing a rapid, cost-effective, and easy-to-use biosensor for early diagnosis of bacterial pathogens. Herein, we developed a simple, label-free, and highly sensitive immunosensor based on electrochemical detection using the Au@MoS₂–PANI nanocomposite. The conductivity of the glassy carbon electrode is greatly enhanced using the Au@MoS₂–PANI nanocomposite and a self-assembled monolayer of mercaptopropionic acid on the gold nanoparticle surface was employed for the covalent immobilization of antibodies to minimize the nonspecific adsorption of bacterial pathogens on the electrode surface. The biosensor established a high selectivity and sensitivity with a low limit of detection of 10 CFU/mL, and detected Escherichia coli within 30 min. Moreover, the developed biosensor demonstrated a good linear detection range, practical utility in urine samples, and electrode regenerative studies.

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