Femtomolar detection of the heart failure biomarker NT-proBNP in artificial saliva using an immersible liquid-gated aptasensor with reduced graphene oxide

Measuring NT-proBNP biomarker is recommended for preliminary diagnostics of the heart failure. Recent studies suggest a possibility of early screening of biomarkers in saliva for non-invasive identification of cardiac diseases at the point-of-care. However, NT-proBNP concentrations in saliva can be thousand time lower than in blood plasma, going down to pg/mL level. To reach this level, we developed a label-free aptasensor based on a liquid-gated field effect transistor using a film of reduced graphene oxide monolayer (rGO-FET) with immobilized NT-proBNP specific aptamer. We found that, depending on ionic strength of tested solutions, there were different levels of correlation in responses of electrical parameters of the rGO-FET aptasensor, namely, the Dirac point shift and transconductance change. The correlation in response to NT-proBNP was high for 1.6 mM phosphate-buffered saline (PBS) and zero for 16 mM PBS in a wide range of analyte concentrations, varied from 1 fg/mL to 10 ng/mL. The effects of transconductance and Dirac point shift in PBS solutions of different concentrations are discussed. The biosensor exhibited a high sensitivity for both transconductance (2 uS/decade) and Dirac point shift (2.3 mV/decade) in diluted PBS with the linear range from 10 fg/mL to 1 pg/mL. The aptasensor performance has been also demonstrated in undiluted artificial saliva with the achieved limit of detection down to 41 fg/mL (~4.6 fM)

Graphene-based field-effect transistor biosensors for the rapid detection and analysis of viruses: A perspective in view of COVID-19

Current situation of COVID-19 demands a rapid, reliable, cost-effective, facile detection strategy to break the transmission chain and biosensor has emerged as a feasible solution for this purpose. Introduction of nanomaterials has undoubtedly improved the performance of biosensor and the addition of graphene enhanced the sensing ability to a peerless level. Amongst different graphene-based biosensing schemes, graphene field-effect transistor marked its unique presence owing to its ability of ultrasensitive and low-noise detection thereby facilitating instantaneous measurements even in the presence of small amounts of analytes. Recently, graphene field-effect transistor type biosensor is even successfully employed in rapid detection of SARS-CoV-2 and this triggers the interest of the scientific community in reviewing the current developments in graphene field-effect transistor. Subsequently, in this article, the recent progress in graphene field-effect transistor type biosensors for the detection of the virus is reviewed and challenges along with their strengths are discussed.Comment: COVID-19, Biosensor, Graphene Field-effect transistor, Virus detectio

THE DESIGN OF A POINT OF CARE FET BIOSENSOR TO DETECT AND SCREEN COVID-19

This work proposes and demonstrates a biosensor with reduced Graphene Oxide (rGO) based Field Effect Transistor (FET) for rapid and selective detection of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). The main objective of this thesis is to detect the SARS-CoV-2 spike protein antigen on spot selectively and rapidly. The rGO channel is coated with the spike protein antibodies to achieve selectivity. Moreover, the biosensing performance and specificity are governed by decorating the sensor’s channel with Metal Nanoparticles (MNPs) such as, copper, and silver. The designed sensor successfully detects the SARS-CoV-2 spike protein and shows singular electrical behavior for detection. The rGO-FET biosensor electronic transport characteristics such as transmission spectrum, electronic current, and transfer curves are studied by using semiempirical modeling combined with a nonequilibrium Green’s function. The transmission spectrum, I-V and transfer curves are investigated to spot the performance alteration caused by detecting the target molecule. The sensor is also tested against another virus, namely Rabies virus, and showed no detection reaction towards it. The introduced sensor is 8.2 nm long and 6.1 nm wide which makes it a perfect candidate for easy handling and transporting. RGO FET-based biosensor is developed and tested to take the advantage of the unique electronic properties of the rGO channel and offer a quick, rapid, easy, and accurate detection method for SARS-CoV-2 virus. The semiempirical study, along with the simulations results are in agreement with the previous literature studies and provide an excellent pathway for practical fabrication

Electronic and electrochemical viral detection for point-of-care use: A systematic review

From PLOS via Jisc Publications RouterHistory: collection 2021, received 2021-07-05, accepted 2021-09-15, epub 2021-09-30Publication status: PublishedFunder: EPSRC Graphene NowNano CDT; Grant(s): EP/L01548X/1Funder: Dame Kathleen Ollerenshaw FellowshipDetecting viruses, which have significant impact on health and the economy, is essential for controlling and combating viral infections. In recent years there has been a focus towards simpler and faster detection methods, specifically through the use of electronic-based detection at the point-of-care. Point-of-care sensors play a particularly important role in the detection of viruses. Tests can be performed in the field or in resource limited regions in a simple manner and short time frame, allowing for rapid treatment. Electronic based detection allows for speed and quantitative detection not otherwise possible at the point-of-care. Such approaches are largely based upon voltammetry, electrochemical impedance spectroscopy, field effect transistors, and similar electrical techniques. Here, we systematically review electronic and electrochemical point-of-care sensors for the detection of human viral pathogens. Using the reported limits of detection and assay times we compare approaches both by detection method and by the target analyte of interest. Compared to recent scoping and narrative reviews, this systematic review which follows established best practice for evidence synthesis adds substantial new evidence on 1) performance and 2) limitations, needed for sensor uptake in the clinical arena. 104 relevant studies were identified by conducting a search of current literature using 7 databases, only including original research articles detecting human viruses and reporting a limit of detection. Detection units were converted to nanomolars where possible in order to compare performance across devices. This approach allows us to identify field effect transistors as having the fastest median response time, and as being the most sensitive, some achieving single-molecule detection. In general, we found that antigens are the quickest targets to detect. We also observe however, that reports are highly variable in their chosen metrics of interest. We suggest that this lack of systematisation across studies may be a major bottleneck in sensor development and translation. Where appropriate, we use the findings of the systematic review to give recommendations for best reporting practice

Polyclonal Aptamer Libraries from a FluRoot-SELEX for the Specific Labeling of the Apical and Elongation/Differentiation Zones of Arabidopsis thaliana Roots

In more than 30 years of aptamer research, it has become widely accepted that aptamers are fascinating binding molecules for a vast variety of applications. However, the majority of targets have been proteins, although special variants of the so-called SELEX process for the molecular evolution of specific aptamers have also been developed, allowing for the targeting of small molecules as well as larger structures such as cells and even cellular networks of human (tumor) tissues. Although the provocative thesis is widely accepted in the field, that is, in principle, any level of complexity for SELEX targets is possible, the number of studies on whole organs or at least parts of them is limited. To pioneer this thesis, and based on our FluCell-SELEX process, here, we have developed polyclonal aptamer libraries against apices and the elongation/differentiation zones of plant roots as examples of organs. We show that dedicated libraries can specifically label the respective parts of the root, allowing us to distinguish them in fluorescence microscopy. We consider this achievement to be an initial but important evidence for the robustness of this SELEX variant. These libraries may be valuable tools for plant research and a promising starting point for the isolation of more specific individual aptamers directed against root-specific epitopes

Novel Electrochemical Biosensors for Clinical Assays

Biosensors, i.e., devices where biological molecules or bio(mimetic)structures are intimately coupled to a chemo/physical transducer for converting a biorecognition event into a measurable signal, have recently gained a wide (if not huge) academic and practical interest for the multitude of their applications in analysis, especially in the field of bioanalysis, medical diagnostics, and clinical assays. Indeed, thanks to their very simple use (permitting sometimes their application at home), the minimal sample pretreatment requirement, the higher selectivity, and sensitivity, biosensors are an essential tool in the detection and monitoring of a wide range of medical conditions from glycemia to Alzheimer’s disease as well as in the monitoring of drug responses. Soon, we expect that their importance and use in clinical diagnostics will expand rapidly so as to be of critical importance to public health in the coming years. This Special Issue would like to focus on recent research and development in the field of biosensors as analytical tools for clinical assays and medical diagnostics

The development of molecularly imprinted polymers for sensor and colorimetric assay applications

The development of devices capable of sensing the presence or absence of molecules is a staple of the modern day world, with fields such as healthcare, forensics, agriculture and industrial processing relying upon biosensors to operate. The presented work sets focus on the use of Molecularly Imprinted Polymers (MIPs) as receptor elements in biosensing applications, demonstrating how these synthetic alternatives to traditional affinity reagents are of value. The thesis initially gives a detailed overview of the current MIP landscape, before determining areas of the field that are currently underdeveloped. The ensuing research highlights how these areas can be built upon, deploying MIPs for drug analysis and antibiotic detection. To this end, the use of MIPs in conjugation with a thermal biosensing platform is presented before shifting the research towards the use of these synthetic receptors in simple colorimetric assays designed for the rapid analysis of compounds

Novel nanoarchitectures for electrochemical biosensing

Thesis advisor: Thomas C. ChilesSensitive, real-time detection of biomarkers is of critical importance for rapid and accurate diagnosis of disease for point-of-care (POC) technologies. Current methods, while sensitive, do not adequately allow for POC applications due to several limitations, including complex instrumentation, high reagent consumption, and cost. We have investigated two novel nanoarchitectures, the nanocoax and the nanodendrite, as electrochemical biosensors towards the POC detection of infectious disease biomarkers to overcome these limitations. The nanocoax architecture is composed of vertically-oriented, nanoscale coaxial electrodes, with coax cores and shields serving as integrated working and counter electrodes, respectively. The dendritic structure consists of metallic nanocrystals extending from the working electrode, increasing sensor surface area. Nanocoaxial- and nanodendritic-based electrochemical sensors were fabricated and developed for the detection of bacterial toxins using an electrochemical enzyme-linked immunosorbent assay (ELISA) and differential pulse voltammetry (DPV). Proof-of-concept was demonstrated for the detection of cholera toxin (CT). Both nanoarchitectures exhibited levels of sensitivity that are comparable to the standard optical ELISA used widely in clinical applications. In addition to matching the detection profile of the standard ELISA, these electrochemical nanosensors provide a simple electrochemical readout and a miniaturized platform with multiplexing capabilities toward POC implementation. Further development as suggested in this thesis may lead to increases in sensitivity, enhancing the attractiveness of the architectures for future POC devices.Thesis (PhD) — Boston College, 2016.Submitted to: Boston College. Graduate School of Arts and Sciences.Discipline: Biology