Advancements in Chronic Myeloid Leukemia detection: Development and evaluation of a novel QCM aptasensor for use in clinical practice.

Oncological diseases represent a significant global health challenge, with high mortality rates. Early detection is crucial for effective treatment, and aptamers, which demonstrate superior specificity and stability compared to antibodies, offer a promising avenue for diagnostic advancement. This study presents the design, development and evaluation of a quartz crystal microbalance (QCM) sensor functionalized with the T2-KK1B10 aptamer for the sensitive and specific detection of Chronic Myeloid Leukemia (CML) K562 cells. The research focuses on optimizing the biorecognition layer by adjusting the aptamer conditions, demonstrating the sensor's ability to detect these CML cells with high specificity and sensitivity. The aptamer-modified QCM sensor operates on the principle of mass change detection upon binding of target cells. By employing the Langmuir isotherm model, the performance of the sensor was optimized for the capture of CML cells from biological samples with LOD of 263 K562 cells. The sensor was also successfully regenerated multiple times without sensitivity loss. Validation of the sensor's performance was conducted under controlled laboratory settings, followed by extensive testing utilizing human lyophilized plasma and clinical samples from patients. The sensor exhibited high sensitivity and specificity in the detection of CML cells within clinical specimens, thereby illustrating its potential for practical clinical deployment. This research presents a novel approach to the early diagnosis of CML, facilitating timely intervention and enhanced patient outcomes. The developed aptasensor demonstrates potential for broader application in cancer diagnostics and personalized medicine.

Rapid and High-Density Antibody Immobilization Using Electropolymerization of Pyrrole for Highly Sensitive Immunoassay.

Polypyrrole (Ppy) is a biologically compatible polymer that is used as a matrix, in which drugs and enzymes can be incorporated by doping. Here, we suggest an inventive application of Ppy as a biorecognition film encapsulated with an antibody (Ab) as an alternative strategy for the on-site multistep functionalization of thiol-based self-assembled monolayers. The fabrication steps of the recognition films were followed by dropping pyrrole and Ab mixed solutions onto the electrode and obtaining a thin film by direct current electropolymerization. The efficiency of Ab immobilization was studied by using fluorescence microscopy and electrochemical (EC) methods. Finally, the Ab density was increased and immobilized in 1 min, and the sensing performance as an EC immunosensor was demonstrated using α-fetoprotein with a limit of detection of 3.13 pg/mL and sensing range from 1 pg/mL to 100 ng/mL. This study demonstrates the potential for electrochemical functionalization of biomolecules with high affinity and rapidity.

Strategy, Design, and Fabrication of Electrochemical Biosensors: A Tutorial.

Advanced healthcare requires novel technologies capable of real-time sensing to monitor acute and long-term health. The challenge relies on converting a real-time quantitative biological and chemical signal into a desired measurable output. Given the success in detecting glucose and the commercialization of glucometers, electrochemical biosensors continue to be a mainstay of academic and industrial research activities. Despite the wealth of literature on electrochemical biosensors, reports are often specific to a particular application (e.g., pathogens, cancer markers, glucose, etc.), and most fail to convey the underlying strategy and design, and if it is transferable to detection of a different analyte. Here we present a tutorial review for those entering this research area that summarizes the basic electrochemical techniques utilized as well as discusses the designs and optimization strategies employed to improve sensitivity and maximize signal output.

Improving synthesis and binding affinities of nucleic acid aptamers and their therapeutics and diagnostic applications.

Nucleic acid aptamers have captivated the attention of analytical and medicinal scientists globally due to their several advantages as recognition molecules over conventional antibodies because of their small size, simple and inexpensive synthesis, broad target range, and high stability in varied environmental conditions. These recognition molecules can be chemically modified to make them resistant to nuclease action in blood serum, reduce rapid renel clearance, improve the target affinity and selectivity, and make them amenable to chemically conjugate with a support system that facilitates their selective applications. This review focuses on the development of efficient aptamer candidates and their application in clinical diagnosis and therapeutic applications. Significant advances have been made in aptamer-based diagnosis of infectious and non-infectious diseases. Collaterally, the progress made in therapeutic applications of aptamers is encouraging, as evident from their use in diagnosing cancer, neurodegenerative diseases, microbial infection, and in imaging. This review also updates the progress on clinical trials of many aptamer-based products of commercial interests. The key development and critical issues on the subject have been summarized in the concluding remarks.

Cutting-edge biorecognition strategies to boost the detection performance of COVID-19 electrochemical biosensors: A review.

Electrochemical biosensors are known for their high sensitivity, selectivity, and low cost. Recently, they have gained significant attention and became particularly important as promising tools for the detection of COVID-19 biomarkers, since they offer a rapid and accurate means of diagnosis. Biorecognition strategies are a crucial component of electrochemical biosensors and determine their specificity and sensitivity based on the interaction of biological molecules, such as antibodies, enzymes, and DNA, with target analytes (e.g., viral particles, proteins and genetic material) to create a measurable signal. Different biorecognition strategies have been developed to enhance the performance of electrochemical biosensors, including direct, competitive, and sandwich binding, alongside nucleic acid hybridization mechanisms and gene editing systems. In this review article, we present the different strategies used in electrochemical biosensors to target SARS-CoV-2 and other COVID-19 biomarkers, as well as explore the advantages and disadvantages of each strategy and highlight recent progress in this field. Additionally, we discuss the challenges associated with developing electrochemical biosensors for clinical COVID-19 diagnosis and their widespread commercialization.

Nanomaterials Connected to Bioreceptors to Introduce Efficient Biosensing Strategy for Diagnosis of the TORCH Infections: A Critical Review.

TORCH infection is a significant risk factor for severe fetal damage, especially congenital malformations. Screening pregnant women for TORCH pathogens could reduce the incidence of adverse pregnancy outcomes and prevent birth defects. Hence, timely identification and inhibition of TORCH infections are effective ways to successfully prevent them in pregnant women. Recently, the superiority of biosensors in TORCH pathogen sensing has been emphasized due to their intrinsic benefits, such as rapid response time, portability, cost-effectiveness, much friendlier preparation and determination steps. With the introduction of advanced nanomaterials into biosensing, the diagnostic properties of biosensors have significantly improved. This study core presents and debates the current progress in biosensing systems for TORCH pathogens using various artificial and natural receptors. The incorporation of nanomaterials into various transduction systems can enhance diagnostic performance. The key performance characteristics of optical and electrochemical biosensors, such as response time, limit of detection (LOD), and linear detection range, are systematically discussed, along with the current TORCH pathogens used for constructing biosensors. Finally, the major problems that exist for converting scientific investigation into product development are also outlined.

Modified Impedance Sensing System Determination of Virulence Characteristics of Pathogenic Bacteria Klebsiella Species.

An impedance sensing system is a family of biosensors that measure changes in electrical impedance to perform their functions. Physical and chemical changes in the impedance of the sensing element, such as changes in the concentration of a target analyte or changes in the physical properties of the sensing element, can result in changes in the impedance of the sensing element. Many impedance biosensors have been developed for the detection of pathogens in the past few decades. Several types of biosensors have been developed for the detection of infections, including transduction elements, biorecognition components, and electrochemical approaches. In this review, we discuss the characteristics and pathogenic factors associated with 2,3-butanediol-producing Klebsiella pneumoniae collected using impedance sensors. An impedance sensing system was introduced as a great method for monitoring the virulence factors of Klebsiella spp. in situ. Klebsiella pneumoniae produces virulence factors, including capsules, lipopolysaccharides, fimbriae, and siderophores, as part of its pathogenesis. It is possible to examine virulence factors' pathogenic characteristics in vitro and in vivo using real tissues or mouse models in order to conduct experiments. For the monitoring of virulence factors in situ, a novel alternative method has been developed to mimic the environment of real tissues. For the purpose of developing tissue-mimicking models, mucin and mannose were used to modify the surface of gold electrodes. These components are known to contribute to the adhesion of pathogens to epithelial cells in mammals.

Peptide-Functionalized Carbon Nanotube Chemiresistors: The Effect of Nanotube Density on Gas Sensing.

Biorecognition element (BRE)-based carbon nanotube (CNT) chemiresistors have tremendous potential to serve as highly sensitive, selective, and power-efficient volatile organic compound (VOC) sensors. While many research groups have studied BRE-functionalized CNTs in material science and device development, little attention has been paid to optimizing CNT density to improve chemiresistor performance. To probe the effect of CNT density on VOC detection, we present the chemiresistor-based sensing results from two peptide-based CNT devices counting more than 60 different individual measurements. We find that a lower CNT density shows a significantly higher noise level and device-to-device variation while exhibiting mildly better sensitivity. Further investigation with SEM images suggests that moderately high CNT density with a stable connection of the nanotube network is desirable to achieve the best signal-to-noise ratio. Our results show an essential design guideline for tuning the nanotube density to provide sensitive and stable chemiresistors.

Feasibility of NAD(P)/NAD(P)H as redox agents in enzymatic plasmonic gold nanostar assays for galactose quantification.

Plasmonic colorimetric sensors have emerged as powerful analytical tools in biochemistry due to their localized surface plasmon resonance extinction in the visible range. Here, we describe the feasibility of NAD(P)/NAD(P)H as redox agents in enzymatic plasmonic gold nanostar (AuNS) assays for galactose quantification using three model enzymes, GalDH, AR and GalOx, immobilized separately on polyvinylpyrrolidone-capped AuNS scaffolds. These highly specific, sensitive and selective bioassays induce the transformation of AuNS into quasi-spherical nanoparticles during the biorecognition of galactose in water and synthetic blood matrices. As a result, using our inexpensive and simple AuNS plasmon bioassays, the presence of galactose may be detected spectrophotometrically and by the naked eye.

Strategies and Applications of Graphene and Its Derivatives-Based Electrochemical Sensors in Cancer Diagnosis.

Graphene is an emerging nanomaterial increasingly being used in electrochemical biosensing applications owing to its high surface area, excellent conductivity, ease of functionalization, and superior electrocatalytic properties compared to other carbon-based electrodes and nanomaterials, enabling faster electron transfer kinetics and higher sensitivity. Graphene electrochemical biosensors may have the potential to enable the rapid, sensitive, and low-cost detection of cancer biomarkers. This paper reviews early-stage research and proof-of-concept studies on the development of graphene electrochemical biosensors for potential future cancer diagnostic applications. Various graphene synthesis methods are outlined along with common functionalization approaches using polymers, biomolecules, nanomaterials, and synthetic chemistry to facilitate the immobilization of recognition elements and improve performance. Major sensor configurations including graphene field-effect transistors, graphene modified electrodes and nanocomposites, and 3D graphene networks are highlighted along with their principles of operation, advantages, and biosensing capabilities. Strategies for the immobilization of biorecognition elements like antibodies, aptamers, peptides, and DNA/RNA probes onto graphene platforms to impart target specificity are summarized. The use of nanomaterial labels, hybrid nanocomposites with graphene, and chemical modification for signal enhancement are also discussed. Examples are provided to illustrate applications for the sensitive electrochemical detection of a broad range of cancer biomarkers including proteins, circulating tumor cells, DNA mutations, non-coding RNAs like miRNA, metabolites, and glycoproteins. Current challenges and future opportunities are elucidated to guide ongoing efforts towards transitioning graphene biosensors from promising research lab tools into mainstream clinical practice. Continued research addressing issues with reproducibility, stability, selectivity, integration, clinical validation, and regulatory approval could enable wider adoption. Overall, graphene electrochemical biosensors present powerful and versatile platforms for cancer diagnosis at the point of care.

Architectural Effect on Self-Assembly and Biorecognition of Randomly Grafted Linear and Branched Polymers at Liquid Crystal-Water Interfaces.

Because of simple synthetic strategies, randomly functionalized amphiphilic polymers have gained much attention. Recent studies have demonstrated that such polymers can be reorganized into different nanostructures, such as spheres, cylinders, vesicles, etc., similar to amphiphilic block copolymers. Our study investigated the self-assembly of randomly functionalized hyperbranched polymers (HBP) and their linear analogues (LP) in solution and at the liquid crystal-water (LC-water) interfaces. Regardless of their architecture, the designed amphiphiles self-assembled into spherical nanoaggregates in solution and mediated the ordering transitions of LC molecules at the LC-water interface. However, the amount of amphiphiles required for LP was 10 times lower than that required for HBP amphiphiles to mediate the same ordering transition of LC molecules. Further, of the two compositionally similar amphiphiles (linear and branched), only the linear architecture responds to biorecognition events. The architectural effect can be attributed to both of these differences mentioned above.

Recent Advances in the Development and Characterization of Electrochemical and Electrical Biosensors for Small Molecule Neurotransmitters.

Neurotransmitters act as chemical messengers, determining human physiological and psychological function, and abnormal levels of neurotransmitters are related to conditions such as Parkinson's and Alzheimer's disease. Biologically and clinically relevant concentrations of neurotransmitters are usually very low (nM), so electrochemical and electronic sensors for neurotransmitter detection play an important role in achieving sensitive and selective detection. Additionally, these sensors have the distinct advantage to potentially be wireless, miniaturized, and multichannel, providing remarkable opportunities for implantable, long-term sensing capabilities unachievable by spectroscopic or chromatographic detection methods. In this article, we will focus on advances in the development and characterization of electrochemical and electronic sensors for neurotransmitters during the last five years, identifying how the field is progressing as well as critical knowledge gaps for sensor researchers.

Electrochemical and Bioelectrochemical Sensing Platforms for Diagnostics of COVID-19.

Rapid transmission and high mortality rates caused by the SARS-CoV-2 virus showed that the best way to fight against the pandemic was through rapid, accurate diagnosis in parallel with vaccination. In this context, several research groups around the world have endeavored to develop new diagnostic methods due to the disadvantages of the gold standard method, reverse transcriptase polymerase chain reaction (RT-PCR), in terms of cost and time consumption. Electrochemical and bioelectrochemical platforms have been important tools for overcoming the limitations of conventional diagnostic platforms, including accuracy, accessibility, portability, and response time. In this review, we report on several electrochemical sensors and biosensors developed for SARS-CoV-2 detection, presenting the concepts, fabrication, advantages, and disadvantages of the different approaches. The focus is devoted to highlighting the recent progress of electrochemical devices developed as next-generation field-deployable analytical tools as well as guiding future researchers in the manufacture of devices for disease diagnosis.

ELISA-Based Biosensors.

Enzyme-linked immunosorbent assay (ELISA) is by definition a biosensor. However, not all immuno-biosensors involve the use of enzymes, while other biosensors incorporate ELISA as a key signaling component. In this chapter, we review the role of ELISA in signal amplification, integration with microfluidic systems, digital labeling, and electrochemical detection.

Direct Identification of Label-Free Gram-Negative Bacteria with Bioreceptor-Free Concentric Interdigitated Electrodes.

This investigation demonstrates an electrochemical method for directly identifying unlabeled Gram-negative bacteria without other additives or labeling agents. After incubation, the bacterial cell surface is linked to the interdigitated electrode through electroadsorption. Next, these cells are exposed to a potential difference between the two electrodes. The design geometry of an electrode has a significant effect on the electrochemical detection of Gram-negative bacteria. Therefore, electrode design geometry is a crucial factor that needs to be considered when designing electrodes for electrochemical sensing. They provide the area for the reaction and are responsible for transferring electrons from one electrode to another. This work aims to study the available design in the commercial market to determine the most suitable electrode geometry with a high detection sensitivity that can be used to identify and quantify bacterial cells in normal saline solutions. To work on detecting bacterial cells without the biorecognition element, we have to consider the microelectrode's design, which makes it very susceptible to bacteria size. The concentration-dilution technique measures the effect of the concentration on label-free Gram-negative bacteria in a normal saline solution without needing bio-recognized elements for a fast screening evaluation. This method's limit of detection (LOD) cannot measure concentrations less than 102 CFU/mL and cannot distinguish between live and dead cells. Nevertheless, this approach exhibited excellent detection performance under optimal experimental conditions and took only a few hours.

Bio-Tailored Sensing at the Nanoscale: Biochemical Aspects and Applications.

The demonstration of the first enzyme-based electrode to detect glucose, published in 1967 by S. J. Updike and G. P. Hicks, kicked off huge efforts in building sensors where biomolecules are exploited as native or modified to achieve new or improved sensing performances. In this growing area, bionanotechnology has become prominent in demonstrating how nanomaterials can be tailored into responsive nanostructures using biomolecules and integrated into sensors to detect different analytes, e.g., biomarkers, antibiotics, toxins and organic compounds as well as whole cells and microorganisms with very high sensitivity. Accounting for the natural affinity between biomolecules and almost every type of nanomaterials and taking advantage of well-known crosslinking strategies to stabilize the resulting hybrid nanostructures, biosensors with broad applications and with unprecedented low detection limits have been realized. This review depicts a comprehensive collection of the most recent biochemical and biophysical strategies for building hybrid devices based on bioconjugated nanomaterials and their applications in label-free detection for diagnostics, food and environmental analysis.

Laser-induced graphene-based electrochemical biosensors for environmental applications: a perspective.

Biosensors are miniaturized devices that provide the advantage of in situ and point-of-care monitoring of analytes of interest. Electrochemical biosensors use the mechanism of oxidation-reduction reactions and measurement of corresponding electron transfer as changes in current, voltage, or other parameters using different electrochemical techniques. The use of electrochemically active materials is critical for the effective functioning of electrochemical biosensors. Laser-induced graphene (LIG) has garnered increasing interest in biosensor development and improvement due to its high electrical conductivity, specific surface area, and simple and scalable fabrication process. The effort of this perspective is to understand the existing classes of analytes and the mechanisms of their detection using LIG-based biosensors. The manuscript has highlighted the potential use of LIG, its modifications, and its use with various receptors for sensing various environmental pollutants. Although the conventional graphene-based sensors effectively detect trace levels for many analytes in different applications, the chemical and energy-intensive fabrication and time-consuming processes make it imperative to explore a low-cost and scalable option such as LIG for biosensors production. The focus of these potential biosensors has been kept on detection analytes of environmental significance such as heavy metals ions, organic and inorganic compounds, fertilizers, pesticides, pathogens, and antibiotics. The use of LIG directly as an electrode, its modifications with nanomaterials and polymers, and its combination with bioreceptors such as aptamers and polymers has been summarized. The strengths, weaknesses, opportunities, and threats analysis has also been done to understand the viability of incorporating LIG-based electrochemical biosensors for environmental applications.

Detection of Emerging Pollutants Using Aptamer-Based Biosensors: Recent Advances, Challenges, and Outlook.

The synergistic potentialities of innovative materials that include aptamers have opened new paradigms in biosensing platforms for high-throughput monitoring systems. The available nucleobase functional moieties in aptamers offer exclusive features for bioanalytical sensing applications. In this context, compared to various in-practice biological recognition elements, the utilization of aptamers in detection platforms results in an extensive range of advantages in terms of design flexibility, stability, and sensitivity, among other attributes. Thus, the utilization of aptamers-based biosensing platforms is extensively anticipated to meet unaddressed challenges of various in-practice and standard analytical and sensing techniques. Furthermore, the superior characteristics of aptasensors have led to their applicability in the detection of harmful pollutants present in ever-increasing concentrations in different environmental matrices and water bodies, seeking to achieve simple and real-time monitoring. Considering the above-mentioned critiques and notable functional attributes of aptamers, herein, we reviewed aptamers as a fascinating interface to design, develop, and deploy a new generation of monitoring systems to aid modern bioanalytical sensing applications. Moreover, this review aims to summarize the most recent advances in the development and application of aptasensors for the detection of various emerging pollutants (EPs), e.g., pharmaceutical, and personal care products (PPCPs), endocrine-disrupting chemicals (EDCs), pesticides and other agricultural-related compounds, and toxic heavy elements. In addition, the limitations and current challenges are also reviewed, considering the technical constraints and complexity of the environmental samples.

Carbon Dots Conjugated Antibody as an Effective FRET-Based Biosensor for Progesterone Hormone Screening.

In this work, carbon dots (CDs) were synthesized by a one-step hydrothermal method using citric acid and ethylene diamine, and covalently functionalized with antibodies for the sensing of progesterone hormone. The structural and morphological analysis reveals that the synthesized CDs are of average size (diameter 8-10 nm) and the surface functionalities are confirmed by XPS, XRD and FT-IR. Further graphene oxide (GO) is used as a quencher due to the fluorescence resonance energy transfer (FRET) mechanism, whereas the presence of the analyte progesterone turns on the fluorescence because of displacement of GO from the surface of CDs effectively inhibiting FRET efficiency due to the increased distance between donor and acceptor moieties. The linear curve is obtained with different progesterone concentrations with 13.8 nM detection limits (R2 = 0.974). The proposed optical method demonstrated high selectivity performance in the presence of structurally resembling interfering compounds. The PL intensity increased linearly with the increased progesterone concentration range (10-900 nM) under the optimal experimental parameters. The developed level-free immunosensor has emerged as a potential platform for simplified progesterone analysis due to the high selectivity performance and good recovery in different samples of spiked water.

Metallic Structures Based on Zinc Oxide Film for Enzyme Biorecognition.

Two structures (Ag/ZnO/ITI/glass: #1 sample and Ag/ZnO/SiO2/Si: #2 sample) are investigated, on the one hand, from the point of view of the formation of evanescent waves in the gratings of metal strips on the structures when the incident TEz wave in the radio frequency range is used. The simulation of the formation of evanescent waves at the edge of the Ag strips, with thicknesses in the range of micrometers, was carried out before the test in the subwavelength regime, with the help of a new improved transducer with metamaterial (MM) lenses. By simulation, a field snapshot was obtained in each sequence of geometry. The evanescent waves are emphasized in the plane XY, due to the scattering of the field on the edge of the strips. On the other hand, ZnO nanoparticles are investigated as a convenient high-efficiency biodetection material, where these structures were used as a biosensitive element to various enzymes (glucose, cholesterol, uric acid, and ascorbic acid). The obtained results demonstrate that the investigated structures based on ZnO nanostructures deposited on different supports are fast and sensitive for enzyme detection and can be successfully incorporated into a device as a biosensing element.