Aptamer Biosensor Research Articles

A dual-mode biosensor featuring single-atom Fe nanozyme for multi-pesticide detection in vegetables

The single-atom iron nanozyme (SA-Fe-NZ) exhibits high catalytic activity and excellent electron transfer efficiency in biosensors. However, the binding of bioreceptors to the surface of SA-Fe-NZ results in a decrease in the catalytic activity of the nanozyme due to its toxic effects. We utilized the toxic effects and excellent electrochemical properties of the SA-Fe-NZ to successfully construct a smartphone-assisted dual-mode biosensor. The complex formed by the binding of organophosphorus pesticides (OPs) to the aptamer exhibited toxic effects and inhibited the catalytic activity of the nanozyme, preventing the colorimetric substrate from being catalyzed. Simultaneously, the aptamers labeled with electrochemical signal molecules approached the electrode surface, causing a change in the electrochemical signal. The results demonstrated that the constructed broad-spectrum aptamer biosensor exhibited a low limit of detection of 3.55 fM and a wide linear range of 10-13-10-2 M, allowing for qualitative and quantitative detection of multiple OPs in vegetables.

A deep learning-enabled smartphone platform for rapid and sensitive colorimetric detection of dimethoate pesticide.

A novel deep learning-enabled smartphone platform is developed to assist a colorimetric aptamer biosensor for fast and highly sensitive detection of dimethoate. The colorimetric determination of dimethoate is based on the specific binding of dimethoate and aptamer, which leads to the aggregation of AuNPs in high-concentration NaCl solution, resulting in an obvious color change from red to blue. This color change provides sufficient data for self-learning enabled by a convolutional neural network (CNN) model, which is established to predict dimethoate concentration based on images acquired from a smartphone. To enhance user-friendliness for non-experts, the CNN model is then embedded into a smartphone app, enabling offline detection of dimethoate pesticide in real environments within just 15min using a pre-configured colorimetric probe. The developed platform exhibits superior performance, achieving a regression coefficient of 0.9992 in the concentration range of 0-10μM. Moreover, the app's performance is found to be consistent with the ELISA kit. These remarkable findings demonstrate the potential of combining colorimetric biosensors with smartphone-based deep learning methods for the development of portable and affordable tools for pesticide detection.

Construction of aptamer sensor based on Au nanozymes for ultrasensitive SERS detection of tobramycin

Tobramycin aptamer was adsorbed on the surface of gold nanoparticles (Au NPs) by van der Waals forces, hydrogen bonds, and other factors, limiting nanozyme peroxidase activity. Because tobramycin has a higher affinity for the tobramycin aptamer, including tobramycin in Au@Apts nanozymes was able to dislodge the aptamer from the surface of Au NPs and restore the catalytic activity of nanozymes. Raman spectroscopy of the enzyme-catalyzed product TMBox can detect this shift in enzyme activity. The restored value of nanozyme activity increased with tobramycin concentration and had a good linear correlation in the concentration range of 10−10∼10−1 M with a detection limit of 2.04 × 10−11 M. Aptamer biosensors are built using the SERS platform, which has the advantages of high sensitivity, high precision, and a wide detection range. It is prospectively employed in food safety and the detection of antibiotic residues.

An ultrasensitive J-shaped optical fiber LSPR aptasensor for the detection of Helicobacter pylori

The development of label-free and sensitive detection of pathogenic bacteria is of great significance for disease prevention and public health protection. In this study, an originally bent structure, named as J-shaped optical fiber probe, was first designed to engineer a localized surface plasmon resonance (LSPR) aptamer biosensor for the rapid and ultrasensitive detection of Helicobacter pylori (H. pylori). The J-shaped optical fiber probe exhibited a significant improvement in refractive index sensitivity (RIS) and LSPR signal response. Meantime, the original sequence of aptamer was truncated in order to effectively capture H. pylori on the optical fiber surface. Besides, a spacer nucleic acid with short stem-loop structure was adopted to control the aptamer density on gold nanoparticles (AuNPs) on the surface of the J-shaped optical fiber probe, which displayed a further enhancement in LSPR signal response. Benefitting from these creative designs, the proposed LSPR biosensor can realize label-free and sensitive detection of H. pylori with a detection limit as low as 45 CFU/mL and a wide linear range from 1.0 × 102 CFU/mL to 1.0 × 108 CFU/mL. At the same time, the sensing strategy can detect the pathogenic bacteria from actual water samples in one step just in 30 min without any sample pretreatment. Due to the advantages of ease-to-preparation, high sensitivity, and rapid analysis, this proposed J-shaped optical fiber LSPR aptasensor can provide a potential strategy for point-of-caring detection of pathogenic bacteria in environmental monitoring and disease diagnosis.

Study on a nano-porous gold/polyamidoamine (NPG/PAMAM)-based electrochemical aptamer biosensor for the detection of ochratoxin a in the red wine

In this study, a novel electrochemical aptamer sensor for detecting ochratoxin A (OTA) was constructed. First, a gold-copper alloy film was prepared via electrochemical deposition, and copper was selectively dissolved in constant potential mode for obtaining the nano-porous gold modified screen-printed carbon electrodes (NPG/SPCE). Then, 2-mercaptoethylamine was dropped on the NPG/SPCE surface and Au-S covalent bonds were formed for immobilizing the metal. Glutaraldehyde as cross-linking agent was added, which resulted in immobilization and attachment of PAMAM to the 2-mercaptoethylamine through the dehydration condensation reaction. During the preparation process, the nano-porous gold and PAMAM-modified layers were characterized by SEM, XRD, and IR spectroscopy, respectively. The characterization results showed that the nano-porous gold and PAMAM composite films were successfully modified. Finally, the OTA aptamer was cross-linked with PAMAM by glutaraldehyde to complete construction of the Apt/PAMAM/NPG/SPCE sensor. The electrochemical performance of this sensor was tested in ochratoxin A solutions with the DPV method. The results showed that the sensor’s reproducibility, stability, and specificity were good. The spiked recoveries in red wine ranged from 99.65%∼101.6%, with a linear range of 0.5 ng/mL∼20 ng/mL and a minimum detection limit of 0.141 ng/mL. Thus, the novel biosensor may provide a promising tool for the trace detection of OTA.

Target-Induced AIE Effect Coupled with CRISPR/Cas12a System Dual-Signal Biosensing for the Ultrasensitive Detection of Gliotoxin.

Here, a novel rapid and ultrasensitive aptamer biosensor was designed for target-induced activation of AIE effect and followed by the activation of Crispr Cas12a (LbCpf1)-mediated cleavage to achieve dual-signal detection. The prepared DNA building blocks contain the target aptamer, ssDNA-Fc, and Activator1. In this system, the activation mode was divided into two steps. First, when the target interacts with the aptamers, the DNA building blocks would be disintegrated rapidly, releasing a mass of Ac1, generating ETTC-dsDNA aggregated to produce a fluorescence signal by the AIE effect. Second, with the release of Ac2, LbCpf1-crRNA was activated, which greatly improves the ssDNA-Fc cleavage efficiency to render signal amplification and ultrasensitive detection of the target. Satisfactorily, using this approach to detect gliotoxin, optimal conditions for detection was achieved for reducing the detection time to 55 min, achieving a low detection limit of 2.4 fM and a satisfactory linear in the range of 50 fM to 1 nM, which addressed the shortcoming of a weak electrochemical signal in previous sensors. The water-insoluble AIE material was coupled with DNA to obtain water-soluble ETTC-dsDNA and successfully introduced into the sensor system, with a low detection limit of 5.6 fM. Subsequently, the biosensor combined with handheld electrochemical workstation was successfully applied in the detection of gliotoxin in five actual samples, with a detection range of 32.0 to 2.09 × 108 pM. This strategy not only provides a novel and effective detection platform for mycotoxins in complex food matrices but also opens a promising avenue for various molecules detection in imaging and disease diagnosis.

Research progress of DNA aptamer-based silver ions detection

Silver ions are regarded as one of the most hazardous metal contaminants, endangering both the ecological environment and human health. Traditional silver ions detection technologies are hampered by their high cost, time-consuming nature, and labor-intensive operation. DNA aptamers play an essential part in the field of biosensors due to their ease of synthesis, ease of modification, and low cost. This paper focuses on reviewing the research progress of DNA aptamer-based biosensors in silver ions detection. According to the types of signal transduction, they are classified into four forms of signal transduction: fluorescent biosensors, colorimetric biosensors, electrochemical biosensors, and surface-enhanced Raman spectroscopy biosensors. In addition, this paper gives a perspective on the application prospects outlook and development directions of DNA aptamer-based biosensors, in order to provide theoretical ideas for the future development of more sensitive DNA aptamer biosensors for rapid detection of silver ions.

Electrochemical aptamer biosensor for DNA detection based on label-free aptamers

Electrochemical aptasensor has been broadly advanced for nucleic acid detection. However, it is a long-term goal to design an aptasensor with high specificity, flexibility, and simplicity. In this work, we develop a strategy of triblock DNA probe, which consists of two DNA probes at both ends and ployA fragments in the middle as probe-polyA-probe. PolyA fragment has high affinity to the surface of gold electrode, so it can be assembled on the electrode surface via polyA instead of traditional Au-S bonds. When the target DNA is simultaneously hybridized with the two capture probes, the hybridization stability can be improved due to the strong base stacking effect. [Ru(NH3)6]3+, as signal probe, can be electrostatically adsorbed on the negatively charged DNA skeleton. A wide linear range (10 pM-10 μM) is obtained with a detection limit of 2.9 pM. Our electrochemical aptasensor has good repeatability, stability, and specificity. More importantly, the electrochemical sensor can successfully detect DNA in human serum samples, which proves its practical value and extensive applicability in complex environment.

Label-Free Electrochemical Aptasensor for Sensitive Detection of Malachite Green Based on AuNPs/MWCNTs@TiO2 Nanocomposites

This study proposes a label-free aptamer biosensor for the sensitive detection of malachite green(MG) using gold nanoparticles/multi-walled carbon nanotubes @ titanium dioxide(AuNPs/MWCNTs@TiO2). The nanocomposite provides a large surface area and good electrical conductivity, improving current transfer and acting as a platform for aptamer immobilization. The aptamer and the complementary chain(cDNA) are paired by base complementary to form the recognition element and fixed on the AuNPs by sulfhydryl group, which was modified on the cDNA. Since DNA is negatively charged, the redox probe in the electrolyte is less exposed to the electrode surface under the repulsion of the negative charge, resulting in a low-electrical signal level. When MG is present, the aptamer is detached from the cDNA and binds to MG, the DNA on the electrode surface is reduced, and the rejection of the redox probe is weakened, which leads to an enhanced electrical signal and enables the detection of MG concentration by measuring the change in the electrical signal. Under the best experimental conditions, the sensor demonstrates a good linear relationship for the detection of MG from 0.01 to 1000 ng/mL, the limit of detection (LOD)is 8.68 pg/mL. This sensor is stable, specific, and reproducible, allowing for the detection of various small-molecule pollutants by changing the aptamer, providing an effective method for detecting small-molecule pollutants.

Electrochemical aptasensors for pathogenic detection toward point-of-care diagnostics.

A biosensor system refers to a biomedical device, which detects biological, chemical, or biochemical components by converting those signals to an electrical signal by utilizing and uniting physical or chemical transducer with biorecognition elements. An electrochemical biosensor is generally based on the reaction of either production or consumption of electrons under a three-electrode system. Biosensor systems are exploited in a wide range of areas, such as medicine, agriculture, husbandry, food, industry, environment protection, quality control, waste disposal, and the military. Pathogenic infections are the third leading cause of death worldwide after cardiovascular diseases and cancer. Therefore, there is an urgent need for effective diagnostic tools to control food, water, and soil contamination result in protecting human life and health. Aptamers are peptide or oligonucleotide-based molecules that show very high affinity to their targets that are produced from large pools of random amino acid or oligonucleotide sequences. Generally, aptamers have been utilized for fundamental sciences and clinical implementations for their target-specific affinity and have been intensely exploited for different kinds of biosensor applications for approximately 30 years. The convergence of aptamers with biosensor systems enabled the construction of voltammetric, amperometric, and impedimetric biosensors for the detection of specific pathogens. In this review, electrochemical aptamer biosensors were evaluated by discussing the definition, types, and production techniques of aptamers, the advantages of aptamers as a biological recognition element against their alternatives, and a wide range of aptasensor examples from literature in the detection of specific pathogens.

Nanomaterial interfaces designed with different biorecognition elements for biosensing of key foodborne pathogens.

Foodborne diseases caused by pathogen bacteria are a serious problem toward the safety of human life in a worldwide. Conventional methods for pathogen bacteria detection have several handicaps, including trained personnel requirement, low sensitivity, laborious enrichment steps, low selectivity, and long-term experiments. There is a need for precise and rapid identification and detection of foodborne pathogens. Biosensors are a remarkable alternative for the detection of foodborne bacteria compared to conventional methods. In recent years, there are different strategies for the designing of specific and sensitive biosensors. Researchers activated to develop enhanced biosensors with different transducer and recognition elements. Thus, the aim of this study was to provide a topical and detailed review on aptamer, nanofiber, and metal organic framework-based biosensors for the detection of food pathogens. First, the conventional methods, type of biosensors, common transducer, and recognition element were systematically explained. Then, novel signal amplification materials and nanomaterials were introduced. Last, current shortcomings were emphasized, and future alternatives were discussed.

Highly Sensitive β-Lactoglobulin Fluorescent Aptamer Biosensors Based on Tungsten Disulfide Nanosheets and DNase I-Assisted Signal Amplification.

β-lactoglobulin (β-Lg) is a protein found in milk that can cause severe allergic reactions, including rash, vomiting, and diarrhea. Thus, it is crucial to develop a sensitive β-Lg detection method to protect people who are susceptible to allergies. Here, we introduce a novel and highly sensitive fluorescent aptamer biosensor for detecting β-Lg. First, a fluorescein-based dye (FAM)-labeled β-lactoglobulin aptamer (β-Lg aptamer) is adsorbed on the surface of tungsten disulfide (WS2) nanosheets via van der Waals forces, resulting in fluorescence quenching. When β-Lg is present, the β-Lg aptamer selectively binds to β-Lg, causing a conformational change in the β-Lg aptamer and releasing it from the surface of WS2 nanosheets, which restores the fluorescence signal. Simultaneously, DNase I in the system cleaves the aptamer bound to the target, producing a short oligonucleotide fragment and releasing β-Lg. The released β-Lg then binds to another β-Lg aptamer adsorbed on WS2, initiating the next round of cleavage, resulting in significant amplification of the fluorescence signal. This method has a linear detection range of 1-100 ng mL-1, and the limit of detection is 0.344 ng mL-1. Furthermore, this approach has been successfully used for detecting β-Lg in milk samples with satisfactory results, providing new opportunities for food analysis and quality control.

Adsorption of tetracycline antibiotics to gold nanoparticles and feasibility of aptamer-based label-free colorimetric detection

Tetracyclines are a group of very important antibiotics that are still in use. To extract, detect, and remove tetracyclines from the environment, various nanomaterials have been employed. Although gold nanoparticles (AuNPs) are a commonly cited material for these purposes, a fundamental understanding of these tetracycline-AuNP systems is still limited. In this work, the adsorption of tetracycline, oxytetracycline, and doxycycline to AuNPs was studied. The effect on the colloidal stability of AuNPs, adsorption kinetics, and the resulting adsorption isotherms was measured. While millimolar concentrations of the tetracyclines can cause aggregation of AuNPs, saturated monolayer adsorption was achieved with low micromolar concentrations of the tetracyclines. Adsorption was instantaneous, and adsorption to AuNPs enhanced their intrinsic fluorescence instead of quenching. With the assumption that aptamer/target complexes cannot be easily adsorbed by AuNPs compared to free aptamers, a label-free colorimetric detection method was tested. While the label-free sensor showed target-dependent aggregation of AuNPs, a nonbinding mutant aptamer showed the same trend, suggesting that the color change did not reflect aptamer adsorption but other events such as target adsorption. This study indicates the importance of the fundamental understanding of target/AuNP interactions to correctly design aptamer and AuNP-based label-free biosensors.

Week-Long Operation of Electrochemical Aptamer Sensors: New Insights into Self-Assembled Monolayer Degradation Mechanisms and Solutions for Stability in Serum at Body Temperature.

Conventional wisdom suggests that widely utilized self-assembled alkylthiolate monolayers on gold are too unstable to last more than several days when exposed to complex fluids such as raw serum at body temperature. Demonstrated here is that these monolayers can not only last at least 1 week under such harsh conditions but that significant applied value can be captured for continuous electrochemical aptamer biosensors. Electrochemical aptamer biosensors provide an ideal tool to investigate monolayer degradation, as aptamer sensors require a tightly packed monolayer to preserve sensor signal vs background current and readily reveal fouling by albumin and other solutes when operating in biofluids. Week-long operation in serum at 37 °C is achieved by (1) increasing van der Waals interactions between adjacent monolayer molecules to increase the activation energy required for desorption, (2) optimizing electrochemical measurement to decrease both alkylthiolate oxidation and electric-field-induced desorption, and (3) mitigating fouling using protective zwitterionic membranes and zwitterion-based blocking layers with antifouling properties. This work further proposes origins and mechanisms of monolayer degradation in a logical stepwise manner that was previously unobservable over multiday time scales. Several of the observed results are surprising, revealing that short-term improvements to sensor longevity (i.e., hours) actually increase sensor degradation in the longer term (i.e., days). The results and underlying insights on mechanisms not only push forward fundamental understanding of stability for self-assembled monolayers but also demonstrate an important milestone for continuous electrochemical aptamer biosensors.

A carbon quantum layer modified BiVO4 photoelectrochemical aptamer biosensor for ultra-sensitive cTnI biomarker detection based on the interface nephelauxetic effect and heterojunction assistance.

The sensitivity and specificity of a semiconductor photoelectrochemical (PEC) aptamer biosensor are determined by the separation and transport of the photoinduced carriers as well as aptamer probe immobilization. In this study, an in situ thermal transformation organic polymer strategy was employed to produce an ∼2.5 nm carbon quantum layer on the surface of the BiVO4(BVO) photoanode. Experimental tests and theoretical calculations have revealed that this carbon quantum layer-coated BVO(C@BVO) heterostructure could generate surface charge depletion regions through an interface nephelauxetic effect. These charge depletion regions facilitated the efficient immobilization of DNA aptamer probes of the acute myocardial infarction biomarker cardiac troponin I (cTnI), while showing almost no immobilization capability on a pure-phase C quantum layer or BVO crystals. Simultaneously, the formation of the C@BVO heterostructure also enhanced the directional transport of photo-generated holes from BVO to the C quantum layer. Due to the above reasons, the C@BVO PEC aptamer biosensor achieved a linear detection range for cTnI from 10-14 g L-1 to 10-10 g L-1, with a record detection limit (LOD) of ∼0.33 × 10-14 g L-1 (N > 3). Meanwhile, the biosensor also demonstrated well the detection reproducibility and specificity for cTnI detection. Therefore, the strategy of using a carbon quantum layer-coated PEC electrode shows good potential to develop PEC biosensors with high sensitivity, specificity, and robustness.

Ultrasensitive detection of CA125 based on a triple signal amplification strategy with a huge number of loaded probes via exonuclease cyclic cleavage, rolling cyclic amplification and strand self-growth.

A novel electrochemiluminescence (ECL) aptamer biosensor with high sensitivity and selectivity for the detection of tumor biomarker carbohydrate antigen 125 (CA125) was constructed, and a strategy of triple amplification of signals was proposed using an exonuclease cyclic cleavage aptamer, combined with rolling ring amplification technologies, generating multi-branched dendritic double-stranded DNA to load a large number of probes through "strand self-growth". The double-stranded DNA, which is abbreviated as CP/CA dsDNA, formed by hybridizing the single strand of capture DNA (CP DNA) with the single strand DNA of the CA125 aptamer (CA Apt) was modified on Fe3O4@Au. When CA125 was added, CP/CA dsDNA was unwound, and CA125 specifically combined with CA Apt to form a protein-aptamer complex, leaving only CP DNA on the surface of Fe3O4@Au. RecJf exonuclease cleaved the aptamer in the protein-aptamer complex and released CA125, which recombined with other CA125 aptamers, to form a cycle that produces more CP DNA on Fe3O4@Au. Three ssDNA (H1, H2, and H3) were introduced and hybridized with CP DNA to form a dsDNA with a "+" configuration structure. Then phi29 DNA polymerase, T4 DNA ligase, deoxy-ribonucleoside triphosphate (dNTP) and padlock probes were added to form a large number of complementary strands of padlock probes (CS padlock probes) by rolling cyclic amplification. CS padlock probes were linked to the "+" type dsDNA; then ssDNA H4 was added and hybridized with the CS padlock probe to form multi-branched dendritic dsDNA. A large number of tris(2,2'-bipyridyl)ruthenium(II) probes were embedded in the double strands, resulting in an extremely strong ECL signal in the presence of the co-reactant tri-n-propylamine (TPA). There is a linear relationship between the ECL signals and CA125 concentrations in the range of 1.0 × 10-15-1.0 × 10-8 mg mL-1, and the detection limit was 2.38 × 10-16 mg mL-1. It has been used for the determination of CA125 in serum samples.

Upconversion nanoparticle-based aptasensor for rapid and ultrasensitive detection of Staphylococcus aureus by low-speed centrifugation.

Opportunistic foodborne pathogens such as Staphylococcus aureus (S. aureus) can cause a wide variety of threats to public health. There is an urgent clinical need for a fast, simple, low-cost, and sensitive method. Here, we designed a fluorescence-based aptamer biosensor (aptasensor) for S. aureus detection using core-shell structured upconversion nanoparticles (CS-UCNPs) as a beacon. A S. aureus-specific aptamer was modified on the surface of CS-UCNPs for binding pathogens. The S. aureus bound to CS-UCNPs can then be isolated from the detection system by simple low-speed centrifugation. Thus, an aptasensor was successfully established for the detection of S. aureus. The fluorescence intensity of CS-UCNPs correlated with the concentration of S. aureus within the range of 6.36 × 102 to 6.36 × 108 CFU mL-1, resulting in the detected limit of S. aureus being 60 CFU mL-1. The aptasensor performed well in real food samples (milk) with a detection limit of 146 CFU mL-1 for S. aureus. Furthermore, we applied our aptasensor in chicken muscles for S. aureus detection, and compared it with the plate count gold standard method. There was no significant difference between our aptasensor and the plate count method within the detected limit, while the time for the aptasensor (0.58 h) was shorter than that of the plate count method (3-4 d). Therefore, we succeeded in the design of a simple, sensitive and fast CS-UCNPs aptasensor for S. aureus detection. This aptasensor system would have the potential for the detection of a wide range of bacterial species by switching the corresponding aptamer.

Aptamers 101: aptamer discovery and in vitro applications in biosensors and separations.

Aptamers are single-stranded nucleic acids that bind and recognize targets much like antibodies. Recently, aptamers have garnered increased interest due to their unique properties, including inexpensive production, simple chemical modification, and long-term stability. At the same time, aptamers possess similar binding affinity and specificity as their protein counterpart. In this review, we discuss the aptamer discovery process as well as aptamer applications to biosensors and separations. In the discovery section, we describe the major steps of the library selection process for aptamers, called systematic evolution of ligands by exponential enrichment (SELEX). We highlight common approaches and emerging strategies in SELEX, from starting library selection to aptamer-target binding characterization. In the applications section, we first evaluate recently developed aptamer biosensors for SARS-CoV-2 virus detection, including electrochemical aptamer-based sensors and lateral flow assays. Then we discuss aptamer-based separations for partitioning different molecules or cell types, especially for purifying T cell subsets for therapeutic applications. Overall, aptamers are promising biomolecular tools and the aptamer field is primed for expansion in biosensing and cell separation.

A novel electrochemical biosensor based on AMNFs@ZIF-67 nano composite material for ultrasensitive detection of HER2

Antimonene (AMNFs) is a new kind of sp2-bonded honeycomb lattice two-dimensional material with strong rod-rail coupling, stability and hydrophilicity. Compared with graphene, antimonene has better sensitivity in DNA molecular sensing. We developed a suitable biosensor-Cd2+-aptamer@AMNFs@ZIF-67 nanocomposite for HER2 biomarker detection. Two-dimensional antimonene can be grown on the surface of ZIF-67 to enhance stability and biocompatibility. The aptamer chain can also be adsorbed on the surface of antimonene, and the complexes of the aptamer and the marker can be detached analytically after targeting the biomarker. The detection limit (LOD) was 4.853 fg/mL within 60 min, the detection range was 0–1000 pg/mL, and the LOD was lower than the existing HER2 aptamer biosensors. The results show that the biosensor has certain applicability and potential, and is expected to be a powerful tool for breast cancer diagnosis in the future.

An ultrasensitive dual-signal ratio electrochemical aptamer biosensor for the detection of HER2

Electrochemical ratiometric biosensors have received attention from researchers because of their self-calibration capability, which can improve the accuracy of detection. In the present study, a ratiometric dual signal electrochemical aptamer biosensor based on ZIF-67 @polydopamine (PDA) nanocomposite and Cu/UiO-66 @ 3, 3′, 5, 5′-tetramethylbenzidine (TMB) nanocomposite was fabricated for the detection of breast cancer biomarker- human epidermal growth factor receptor 2 (HER2). PDA was chosen as the electroactive material with electrochemical redox activity and ZIF-67 with a high specific surface area, forming the ZIF-67 @PDA+Apt as the nanoprobe for capturing HER2 in this paper. Cu/UiO-66 is a bimetallic compound with high stability, specific surface area, and strong adsorption onto aptamer chains, and the Cu/UiO-66 @TMB+Apt nanocomposite was used as a probe for signal labeling. In the presence of HER2, the capture of HER2 by the ZIF-67 @PDA+Apt probe results in a weakening of the conductivity of the electrode, however, by attenuating the electrochemical signal from the PDA, altering the probe-Cu/UiO-66 @TMB+Apt signaling will result in the enhancement of the TMB electrochemical signal. With a sensitive detection of HER2 biomarkers in as little as 30 min with the detection range of 0.75–40 pg/mL and a limit of detection as low as 44.8 fg/mL. Dual signal ratio biosensors have a low limit of detection, short detection time, which can accurately detect targets in complex biological samples, which has important theoretical importance.