Biosensor System Research Articles

Direct Laser Processing and Functionalizing PI/PDMS Composites for an On-Demand, Programmable, Recyclable Device Platform.

Skin-interfaced high-sensitive biosensing systems to detect electrophysiological and biochemical signals have shown great potential in personal health monitoring and disease management. However, the integration of 3D porous nanostructures for improved sensitivity and various functional composites for signal transduction/processing/transmission often relies on different materials and complex fabrication processes, leading to weak interfaces prone to failure upon fatigue or mechanical deformations. The integrated system also needs additional adhesive to strongly conform to the human skin, which can also cause irritation, alignment issues, and motion artifacts. This work introduces a skin-attachable, reprogrammable, multifunctional, adhesive device patch fabricated by simple and low-cost laser scribing of an adhesive composite with polyimide powders and amine-based ethoxylated polyethylenimine dispersed in the silicone elastomer. The obtained laser-induced graphene in the adhesive composite can be further selectively functionalized with conductive nanomaterials or enzymes for enhanced electrical conductivity or selective sensing of various sweat biomarkers. The possible combination of the sensors for real-time biofluid analysis and electrophysiological signal monitoring with RF energy harvesting and communication promises a standalone stretchable adhesive device platform based on the same material system and fabrication process.

Recent Advances in Gold Nanocluster-Based Biosensing and Therapy: A Review.

Gold nanoclusters (Au NCs) with bright emission and unique chemical reactivity characters have been widely applied for optical sensing and imaging. With a combination of surface modifications, effective therapeutic treatments of tumors are realized. In this review, we summarize the recently adopted biosensing and therapy events based on Au NCs. Homogeneous and fluorometric biosensing systems toward various targets, including ions, small molecules, reactive oxygen species, biomacromolecules, cancer cells, and bacteria, in vitro and in vivo, are presented by turn-off, turn-on, and ratiometric tactics. The therapy applications are concluded in three aspects: photodynamic therapy, photothermal therapy, and as a drug carrier. The basic mechanisms and performances of these systems are introduced. Finally, this review highlights the challenges and future trend of Au NC-based biosensing and therapy systems.

Optimizing single-atom cerium nanozyme activity to function in a sequential catalytic system for colorimetric biosensing

Ever-increasing attention is being drawn towards single-atom nanozymes (SAzymes) for colorimetric biosensing applications. A series of atomically dispersed cerium (Ce) nanozymes are synthesized with different feeding molar ratios of Ce/Zn or under various pyrolysis temperatures, and their enzymatic performance, including peroxidase-, oxidase-, and catalase-activities, is explored. Both theoretical and experimental results demonstrate that Ce species symmetrically coordinate with nitrogen atoms to form CeN4 moieties which exhibit peroxidase-like activity. The best Ce-based SAzyme achieves remarkable peroxidase-like specificity and enables the colorimetric sensing of H2O2. This sensitivity provides a versatile reporter platform for detecting the activities of natural enzymes involved in H2O2 generation and further monitoring their substrates as well as inhibitors, for example, organophosphorus pesticides (OPs) when integrating with the acetylcholine esterase-choline oxidase biocatalyzed cascade. More importantly, this proposed sequential catalytic biosensor exhibits the capability to monitor trace OPs in the real samples with high sensitivity yet insignificant interference. This work sheds new light on further design of highly active lanthanide-based SAzymes to boost enzyme-mimicking reactions for the applications in rapid on-site colorimetric analysis of emerging contaminants in environmental and food samples.

Next Generation Chemiluminescent Probes for Antimalarial Drug Discovery.

Malaria is caused by parasites of the Plasmodium genus and remains one of the most pressing human health problems. The spread of parasites resistant to or partially resistant to single or multiple drugs, including frontline antimalarial artemisinin and its derivatives, poses a serious threat to current and future malaria control efforts. In vitro drug assays are important for identifying new antimalarial compounds and monitoring drug resistance. Due to its robustness and ease of use, the [3H]-hypoxanthine incorporation assay is still considered a gold standard and is widely applied, despite limited sensitivity and the dependence on radioactive material. Here, we present a first-of-its-kind chemiluminescence-based antimalarial drug screening assay. The effect of compounds on P. falciparum is monitored by using a dioxetane-based substrate (AquaSpark β-D-galactoside) that emits high-intensity luminescence upon removal of a protective group (β-D-galactoside) by a transgenic β-galactosidase reporter enzyme. This biosensor enables highly sensitive, robust, and cost-effective detection of asexual, intraerythrocytic P. falciparum parasites without the need for parasite enrichment, washing, or purification steps. We are convinced that the ultralow detection limit of less than 100 parasites of the presented biosensor system will become instrumental in malaria research, including but not limited to drug screening.

Enhancing Glucose Biosensing with Graphene Oxide and Ferrocene-Modified Linear Poly(ethylenimine).

We designed and optimized a glucose biosensor system based on a screen-printed electrode modified with the NAD-GDH enzyme. To enhance the electroactive surface area and improve the electron transfer efficiency, we introduced graphene oxide (GO) and ferrocene-modified linear poly(ethylenimine) (LPEI-Fc) onto the biosensor surface. This strategic modification exploits the electrostatic interaction between graphene oxide, which possesses a negative charge, and LPEI-Fc, which is positively charged. This interaction results in increased catalytic current during glucose oxidation and helps improve the overall glucose detection sensitivity by amperometry. We integrated the developed glucose sensor into a flow injection (FI) system. This integration facilitates a swift and reproducible detection of glucose, and it also mitigates the risk of contamination during the analyses. The incorporation of an FI system improves the efficiency of the biosensor, ensuring precise and reliable results in a short time. The proposed sensor was operated at a constant applied potential of 0.35 V. After optimizing the system, a linear calibration curve was obtained for the concentration range of 1.0-40 mM (R2 = 0.986). The FI system was successfully applied to determine the glucose content of a commercial sports drink.

BSA nanoclusters-based sensor for detection of dopamine in schizophrenia from biofluids

Objective To develop nontoxic and stable fluorescent emission B-Cu nanoclusters (NCs) for the specific detection of dopamine at low concentrations in cerebrospinal fluid (CSF). Significance Fluorescent gold and copper NCs conjugated with proteins, such as bovine serum albumin (BSA), offer photostability and healthcare potential. This study focused on fabricating B-Cu NCs that exhibited superior characteristics for sensitive dopamine detection. Methods The study employed various instrumental techniques including attenuated total reflectance Fourier-transform infrared spectroscopy (ATR-FTIR), scanning electron microscopy (SEM), spectrofluorometry, and transmission electron microscopy (TEM) to characterize the formulated B-Cu NCs. The NCs were synthesized, resulting in particle size ∼300 nm. The highest observed fluorescence was recorded at 24542.81 relative fluorescence units (RFU). Results The introduction of dopamine at concentrations of 0.1, 0.2, 0.3, and 0.4 ng/mL led to decreased fluorescence in both B-Au and B-Cu NCs due to an electron transport system. This reduction in fluorescence allowed dopamine concentration analysis in phosphate buffer and biological fluids such as blood plasma and CSF. B-Cu NCs showed potential as a biosensing system for point-of-care (POC) applications, specifically for diagnosing schizophrenia. Conclusion The study successfully synthesized stable and nontoxic B-Cu NCs with enhanced fluorescent emission properties. These NCs exhibited the capacity to detect dopamine at low concentrations in CSF. The study’s findings hold promise for future applications, particularly in the development of a B-Cu NCs-based biosensing system for convenient POC detection of schizophrenia by both patients and clinicians. The potential impact of this technology on healthcare and biomedical fields is substantial.

Development of a Smartphone-linked Optical Biosensing System for Point-of-care Testing Applications

Development of a Smartphone-linked Optical Biosensing System for Point-of-care Testing Applications

A photoelectrochemical aptasensor based on double Z-scheme α-Fe2O3/MoS2/Bi2S3 ternary heterojunction for sensitive detection of circulating tumor cells.

A novel photoelectrochemical (PEC) aptasensor based on a dual Z-scheme α-Fe2O3/MoS2/Bi2S3 ternary heterojunction for the ultrasensitive detection of circulating tumor cells (CTCs) was developed. The α-Fe2O3/MoS2/Bi2S3 nanocomposite was prepared via a step-by-step route, and the photoproduced electron/hole transfer path was speculated by conducting trapping experiments of reactive species. α-Fe2O3/MoS2/Bi2S3-modified electrodes exhibited greatly enhanced photocurrent under visible light due to the double Z-scheme charge transfer process, which met the requirement of the PEC sensor for detecting larger targets. After the aptamer was conjugated on the photoelectrode through chitosan (CS) and glutaraldehyde (GA), when MCF-7 cells were presented and captured, the photocurrent of the PEC biosensing system decreased due to steric hindrance. The current intensity had a linear relationship with the logarithm of MCF-7 cell concentration ranging from 10 to 1×105 cellsmL-1, with a low detection limit of 3 cellmL-1 (S/N = 3). The dual Z-scheme α-Fe2O3/MoS2/Bi2S3 ternary heterojunction-modified PEC aptasensor exhibited high sensitivity and excellent specificity and stability. Additionally, MCF-7 cells in human serum were determined by this PEC aptasensor, exhibiting great potential as a promising tool for clinical detection.

THERMOREGULATION SYSTEM FOR BIOSENSORS BASED ON FIELD-EFFECT TRANSISTORS WITH A NANOWIRE CHANNEL

In this work we present an automatic thermoregulation system for biosensors based on field-effect transistors with a nanowire channel, which provides full control on the required temperature regime in bioanalytical analises. The system elements, including field-effect transistors with a nanowire channel, temperature sensors and heaters, were fabricated on a single silicon cristal using electron beam lithography, reactive ion etching and high-vacuum deposition techniques. Unicue electronics have been developed to control and maintain temperature. The dependence of thermometer readout on heating power was measured, which is in good agreement with the results of numerical simulation. A demonstration of a thermoregulation system with PID-feedback was carried out, ensuring the establishment of a desiered temperature in the range of 30-70◦ C in 18 s in liquid. A demonstration of a thermoregulation system for detecting nucleic acids was carried out using synthetic single-stranded DNA, which is a gene fragment from the bacterium Escherichia coli. The minimal detectable response was observed for a sample with a concentration of 3 fM.

Recent Advances in Nanomaterials Based Optical Sensors for the Detection of Melatonin and Serotonin.

In this review paper we discussed the detection of melatonin and serotonin by using various optical methods. Melatonin and serotonin are very necessary body hormones these are also called neuroregulatory hormones secreted by pineal gland in brain by pinealocytes and shape of pineal gland is cone like. Sensitive detection of melatonin and serotonin in pharmacological samples and human serum is crucial for human beings, lots of research publications available in literature for melatonin and serotonin and we overviewed these papers. We have deeply reviewed many research papers where sensitively sensing of melatonin and serotonin occurs, by using of various interfering agents and nanomaterials. This review aims presenting colorimetry, fluorometry and spectrophotometric detection of melatonin (MEL) and serotonin (SER) by using different metal oxides, carbon nanomaterials (nanosheets, nanorods, nanofibers) and many other agents. Nanomaterials typically possess favourable optical, electrical and mechanical characteristics, they provide up new avenues for enhancing the efficacy of sensors. It is crucial to provide an optical sensors platform that is dependable, sensitive and low price. The development of sensors and biosensors to use nanomaterials for neurotransmitters has advanced significantly in recent years. There are currently many developing biomarkers in biological fluids, and bionanomaterial-based biosensor systems, as well as clinical and pharmacological settings, have garnered significant interest. Biomarkers have been found using optical devices in a quick, selective and sensitive manner. Our aim is to compile all the data that already published on MEL, SER sensing and comparison of each method, we mainly focused on principle, observations, sensitivity, selectivity, limit of detection, mechanism behind the reaction, effect of temperature, pH and concentration. In the last of this paper, we discuss some challenges of these methods and future projects.

3D-printed biosensors in biomedical applications exploiting plasmonic phenomena and antibody self-assembled monolayers.

In this work, a 3D-printed plasmonic chip based on a silver-gold bilayer was developed in order to enhance the optical response of the surface plasmon resonance (SPR) probe. More specifically, numerical and experimental results were obtained on the 3D-printed SPR platform based on a silver-gold bilayer. Then, the optimized probe's gold plasmonic interface was functionalized with a specific antibody directed against the p27Kip1 protein (p27), an important cell cycle regulator. The 3D-printed plasmonic biosensor was tested for p27 detection with good selectivity and a detection limit of 55 pM. The biosensor system demonstrated performance similar to commercially available ELISA (enzyme-linked immunoassay) kits, with several advantages, such as a wide detection range and a modular and simple-based architecture. The proposed biosensing technology offers flexible deployment options that are useful in disposable, low-cost, small-size, and simple-to-use biochips, envisaging future applications in experimental and biomedical research.

System for classifying antibody concentration against severe acute respiratory syndrome coronavirus 2 S1 spike antigen with automatic quick response generation for integration with health passports

Aim: After the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic and the realization of mass vaccination against the virus, the availability of a reliable, rapid, and easy-to-use system for registering the individual anti-S1 antibody titer could facilitate the personalized assessment of the need for booster vaccine doses and the reduction of social distancing and other measures. Methods: The biosensor system is based on immobilized engineered SK-N-SH neuroblastoma cells, bearing the S1 protein, and it can detect immunoglobulin G (IgG) antibodies against the SARS-CoV-2 S1 spike antigen. A disposable electrode strip bearing the engineered mammalian cells is connected to a customized read-out potentiometric device with real-time data transmission to a wireless fidelity (WiFi)-connected smartphone. Blood samples from past-infected individuals and individuals vaccinated against SARS-CoV-2 were used for validation. Results: In the present study, a smartphone application (app), capable of analyzing data regarding the levels of anti-S1 antibodies in blood is introduced. The app works in conjunction with a portable, ultra-rapid, and sensitive biosensor transmitting real-time measurements to the smartphone. Both historical and current individual data can be encoded by using the app, resulting in a widely accepted quick response (QR) code, which can then be constantly updated to match a person’s status. Conclusions: This novel system could be utilized for the eventual development of a coronavirus disease 2019 (COVID-19) electronic passport, which could be further employed to improve the population-wide, cross-country surveillance of vaccination efficiency, as well as facilitate the implementation of cross-border digital health services in a user-friendly and secure way.

Recent advances in smart wearable sensors for continuous human health monitoring

In recent years, the biochemical and biological research areas have shown great interest in a smart wearable sensor because of its increasing prevalence and high potential to monitor human health in a non-invasive manner by continuous screening of biomarkers dispersed throughout the biological analytes, as well as real-time diagnostic tools and time-sensitive information compared to conventional hospital-centered system. These smart wearable sensors offer an innovative option for evaluating and investigating human health by incorporating a portion of recent advances in technology and engineering that can enhance real-time point-of-care-testing capabilities. Smart wearable sensors have emerged progressively with a mixture of multiplexed biosensing, microfluidic sampling, and data acquisition systems incorporated with flexible substrate and bodily attachments for enhanced wearability, portability, and reliability. There is a good chance that smart wearable sensors will be relevant to the early detection and diagnosis of disease management and control. Therefore, pioneering smart wearable sensors into reality seems extremely promising despite possible challenges in this cutting-edge technology for a better future in the healthcare domain. This review presents critical viewpoints on recent developments in wearable sensors in the upcoming smart digital health monitoring in real-time scenarios. In addition, there have been proactive discussions in recent years on materials selection, design optimization, efficient fabrication tools, and data processing units, as well as their continuous monitoring and tracking strategy with system-level integration such as internet-of-things, cyber-physical systems, and machine learning algorithms.

Flow-Through Amperometric Biosensor System Based on Functionalized Aryl Derivative of Phenothiazine and PAMAM-Calix-Dendrimers for the Determination of Uric Acid.

A flow-through biosensor system for the determination of uric acid was developed on the platform of flow-through electrochemical cell manufactured by 3D printing from poly(lactic acid) and equipped with a modified screen-printed graphite electrode (SPE). Uricase was immobilized to the inner surface of a replaceable reactor chamber. Its working volume was reduced to 10 μL against a previously reported similar cell. SPE was modified independently of the enzyme reactor with carbon black, pillar[5]arene, poly(amidoamine) dendrimers based on the p-tert-butylthiacalix[4]arene (PAMAM-calix-dendrimers) platform and electropolymerized 3,7-bis(4-aminophenylamino) phenothiazin-5-ium chloride. Introduction of the PAMAM-calix-dendrimers into the electrode coating led to a fivefold increase in the redox currents of the electroactive polymer. It was found that higher generations of the PAMAM-calix-dendrimers led to a greater increase in the currents measured. Coatings consisted of products of the electropolymerization of the phenothiazine with implemented pillar[5]arene and PAMAM-calix-dendrimers showing high efficiency in the electrochemical reduction of hydrogen peroxide that was formed in the enzymatic oxidation of uric acid. The presence of PAMAM-calix-dendrimer G2 in the coating increased the redox signal related to the uric acid assay by more than 1.5 times. The biosensor system was successfully applied for the enzymatic determination of uric acid in chronoamperometric mode. The following optimal parameters for the chronoamperometric determination of uric acid in flow-through conditions were established: pH 8.0, flow rate 0.2 mL·min-1, 5 U of uricase per reactor. Under these conditions, the biosensor system made it possible to determine from 10 nM to 20 μM of uric acid with the limit of detection (LOD) of 4 nM. Glucose (up to 1 mM), dopamine (up to 0.5 mM), and ascorbic acid (up to 50 μM) did not affect the signal of the biosensor toward uric acid. The biosensor was tested on spiked artificial urine samples, and showed 101% recovery for tenfold diluted samples. The ease of assembly of the flow cell and the low cost of the replacement parts make for a promising future application of the biosensor system in routine clinical analyses.

Label-Free Electrochemical Immunosensor Based on Conjugated Polymer Film Coated Disposable Electrode for Ultrasensitive Determination of Resistin Potential Obesity Biomarker.

A new label-free immunosensor was designed for sensitive detection of resistin obesity biomarker in human biological fluids. To construct a sensing interface, the monomer of double epoxy groups-substituted thiophene (TdiEpx) was synthesized for the fabrication of the biosensing system. A disposable indium tin oxide sheet was first modified by electrochemical polymerization of the TdiEpx monomer, and this robust and novel surface was characterized using different spectroscopic and electrochemical analyses. The double epoxy ends were linked to the amino ends of anti-resistin, and they served as binding points for the covalent binding of biomolecules. The double epoxy ends present in each TdiEpx monomer ensured an extensive surface area, which improved the quantity of attached anti-resistin. The determination of resistin antigen was based on the specific coupling of resistin with anti-resistin, and this interaction hindered the electron transfer reaction. The immunosensor introduced a wide linear range of 0.0125-15 pg/mL, a low detection limit of 4.17 fg/mL, and an excellent sensitivity of 1.38 kohm pg mL-1 cm2. In this study, a sandwich enzyme-linked immunosorbent assay spectrophotometric method was utilized as a reference technique for the quantitative analysis of resistin in human serum and saliva samples. Both measurements in clinical samples displayed correlations and high-correlation coefficients. In addition, this immunosensor had good storage stability, acceptable repeatability and reproducibility, high specificity, and good accuracy. The proposed immunosensor provided a simple and versatile impedimetric immunosensing platform and a promisingly sensitive way for clinical applications.

DNA Cryogels with Anisotropic Mechanical and Responsive Properties for Specific Cell Capture and Release.

Due to their programmable stimuli-responsiveness, excellent biocompatibility, and water-rich and soft structures similar to biological tissues, smart DNA hydrogels hold great promise for biosensing and biomedical applications. However, most DNA hydrogels developed to date are composed of randomly oriented and isotropic polymer networks, and the resulting slow response to biotargets and lack of anisotropic properties similar to those of biological tissues have limited their extensive applications. Herein, anisotropic DNA hydrogels consisting of unidirectional void channels internally oriented up to macroscopic length scales were constructed by a directional cryopolymerization method, as exemplified by a DNA-incorporated covalently cross-linked DNA cryogel and a DNA duplex structure noncovalently cross-linked DNA cryogel. Results showed that the formation of unidirectional channels significantly improved the responsiveness of the gel matrix to biomacromolecular substances and further endowed the DNA cryogels with anisotropic properties, including anisotropic mechanical properties, anisotropic swelling/shrinking behaviors, and anisotropic responsiveness to specific biotargets. Moreover, the abundant oriented and long macroporous channels in the gel matrix facilitated the migration of cells, and through the introduction of aptamer structures and thermosensitive polymers, an anisotropic DNA cryogel-based platform was further constructed to achieve the highly efficient capture and release of specific cells. These anisotropic DNA hydrogels may provide new opportunities for the development of anisotropic separation and biosensing systems.

Biosensing Platforms for Cardiac Biomarker Detection.

Myocardial infarction (MI) is a cardiovascular disease that occurs when there is an elevated demand for myocardial oxygen as a result of the rupture or erosion of atherosclerotic plaques. Globally, the mortality rates associated with MI are steadily on the rise. Traditional diagnostic biomarkers employed in clinical settings for MI diagnosis have various drawbacks, prompting researchers to investigate fast, precise, and highly sensitive biosensor platforms and technologies. Biosensors are analytical devices that combine biological elements with physicochemical transducers to detect and quantify specific compounds or analytes. These devices play a crucial role in various fields including healthcare, environmental monitoring, food safety, and biotechnology. Biosensors developed for the detection of cardiac biomarkers are typically electrochemical, mass, and optical biosensors. Nanomaterials have emerged as revolutionary components in the field of biosensing, offering unique properties that significantly enhance the sensitivity and specificity of the detection systems. This review provides a comprehensive overview of the advancements and applications of nanomaterial-based biosensing systems. Beginning with an exploration of the fundamental principles governing nanomaterials, we delve into their diverse properties, including but not limited to electrical, optical, magnetic, and thermal characteristics. The integration of these nanomaterials as transducers in biosensors has paved the way for unprecedented developments in analytical techniques. Moreover, the principles and types of biosensors and their applications in cardiovascular disease diagnosis are explained in detail. The current biosensors for cardiac biomarker detection are also discussed, with an elaboration of the pros and cons of existing platforms and concluding with future perspectives.

Wireless and battery-free wearable biosensing of riboflavin in sweat for precision nutrition

Nutrition assessment is crucial for dietary guidance and prevention of malnutrition. Recent endeavors in wearable biochemical sensors have enabled real-time, in situ analysis of nutrients in sweat. However, the monitoring of riboflavin, an indispensable vitamin B involved in energy metabolism, remains challenging due to its trace level and variations in the sweat matrix. Herein, we report a wireless, battery-free, and flexible wearable biosensing system for the in situ monitoring of sweat riboflavin. Highly sensitive and selective electrochemical voltammetric detection is realized based on the synergistic effect of electrodeposited reduced graphene oxide (rGO) and platinum nanoparticles (PtNPs) with a low detection limit of 1.2 nM. The fully integrated system is capable of sweat sampling with the microfluidic patch, real-time riboflavin analysis and pH calibration with the flexible electrode array, as well as wirelessly simultaneous near field communication (NFC) energy harvesting and data transmission with the flexible circuit and a smartphone. On-body human sweat analysis demonstrates high accuracy cross-validated with gold-standard measurements, and reveals a strong correlation between sweat and urine riboflavin levels. The proposed wearable platform opens up attractive possibilities for noninvasive nutrient tracking, providing strong potential for personalized dietary guidance towards precision nutrition.

Insertable Biosensors: Combining Implanted Sensing Materials with Wearable Monitors.

Insertable biosensor systems are medical diagnostic devices with two primary components: an implantable biosensor within the body and a wearable monitor that can remotely interrogate the biosensor from outside the body. Because the biosensor does not require a physical connection to the electronic monitor, insertable biosensor systems promise improved patient comfort, reduced inflammation and infection risk, and extended operational lifetimes relative to established percutaneous biosensor systems. However, the lack of physical connection also presents technical challenges that have necessitated new innovations in developing sensing chemistries, transduction methods, and communication modalities. In this review, we discuss the key developments that have made insertables a promising option for longitudinal biometric monitoring and highlight the essential needs and existing development challenges to realizing the next generation of insertables for extended-use diagnostic and prognostic devices.

Unraveling the interplay of DNAzyme and interfacial factors for enhanced biosensing

Deoxyribozyme (DNAzyme) and its substrate hybridization are crucial for achieving desirable detection performance in the DNAzyme coupling nanomaterial biosensor system. However, interfacial factors such as electrostatic repulsion, steric hindrance, and nonspecific adsorption from gold nanoparticles make this hybridization process complicated and challenging. Moreover, the DNAzyme structure changes with different application purposes, which might affect the DNAzyme and substrate's connection. Few studies have focused on the interplay of DNAzyme and interfacial factors in the biosensor field. In this work, three types of DNAzyme variants were designed, and their biosensor performance rules were studied and summarized with the synergistic effect of interfacial factors. Additionally, corresponding biosensor applications, such as multiple modulation functions and miRNA detections, were constructed based on the distinct principles of DNAzyme variants.