{"paper_id":"2768641b-8d2f-4183-8f65-21c5294bfc06","body_text":"Novel Ti3C2Tx MXene Nanozyme with Manageable Catalytic Activity and Application to Electrochemical Biosensor | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Novel Ti3C2Tx MXene Nanozyme with Manageable Catalytic Activity and Application to Electrochemical Biosensor Rongjun Yu, Jian Xue, Yang Wang, Jingfu Qiu, Xinyi Huang, Anyi Chen, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-965061/v2 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 09 Mar, 2022 Read the published version in Journal of Nanobiotechnology → Version 2 posted 6 You are reading this latest preprint version Show more versions Abstract In this work, Ti 3 C 2 T x MXene was identified as efficient nanozyme with area-dependent electrocatalytic activity in oxidation of phenolic compounds, which originated from the strong adsorption effect between the phenolic hydroxyl group and the oxygen atom on the surface of Ti 3 C 2 T x MXene flake. On the basis of the novel electrocatalytic activity, Ti 3 C 2 T x MXene was combined with alkaline phosphatase to construct a novel cascading catalytic amplification strategy using 1-naphthyl phosphate (1-NPP) as substrate, thereby realizing efficient electrochemical signal amplification. Taking advantage of the novel cascading catalytic amplification strategy, an electrochemical biosensor was fabricated for BCR/ABL fusion gene detection, which achieved excellent sensitivity with linear range from 0.2 fM to 20 nM and limit of detection down to 0.05 fM. This biosensor provided a promising tool for ultrasensitive fusion gene detection in early diagnosis of chronic myelogenous leukemia and acute lymphocytic leukemia. Moreover, the manageable catalytic activity of MXene broke a path for developing nanozymes, which possessed enormous application potential in not only electrochemical analysis but also the extensive fields including organic synthesis, pollutant disposal and so on. Ti3C2Tx MXene Nanozyme Electrocatalysis Cascading catalytic amplification Electrochemical biosensor Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 1. Introduction Electrochemical biosensor has become one of the most predominant analysis tools in clinical diagnosis due to the outstanding merits of high sensitivity and selectivity, rapid response, low cost, simple instrumentation, easy miniaturization and good quantitative ability [ 1 – 4 ]. Enzymatic electrocatalysis is a widely used technique in electrochemical biosensor, which improves the analytical sensitivity by promoting the electrochemical redox reactions with the help of enzymes [ 5 – 7 ]. Nevertheless, natural enzymes are general cost to manufacture and store, unstable to transfer or modify, and sensitive to harsh physiochemical conditions. Nanozymes, nanomaterials with enzyme-like catalytic activities, well overcome the shortages of natural enzymes [ 8 , 9 ]. Therefore, nanozymes are attracting increasing attention in bioanalysis to substitute conventional natural enzyme [ 10 , 11 ]. However, the catalytic activities of conventional nanozymes strongly rely on the defect dependent active centers, such as surface dangling bonds or heterogeneous doping domains, which is distributed unevenly on the surface [ 12 , 13 ]. As a consequence, the homogeneity of particle is a very important parameter of nanozymes, which brings great challenge to prepare high-quality product and applied to precise quantitative analysis [ 10 ]. From this prospective, developing nanomaterials with enzyme-like catalytic activities independent to the morphology or crystal defect is of great significance as this kind of materials might be more easily controlled to obtain uniform catalysis activity. MXene is termed for a series of two-dimension (2D) transition metal carbides, nitrides, and carbonitrides [ 14 , 15 ]. Recent years, because typical MXene possess biocompatibility, large specific surface area, rich surface chemistry, tunable lateral size, good electrical conductivity, and mechanical robustness, allowing efficient and selective interaction with target species. MXene-based materials have been utilized as electrocatalysts for detecting small molecules, pharmaceutical drugs, environmental pollutants, and biomarkers. These characteristics render MXene an ideal platform as sensing materials for electrochemical application [ 16 ]. The electrocatalytic activity MXene have attracted enormous research interest in diverse fields, such as hydrogen evolution [ 17 ], oxygen evolution [ 18 , 19 ], N 2 -to-NH 3 conversion [ 20 ], fuel cell [ 21 , 22 ], energy storage [ 23 , 24 ] and carbon dioxide reduction [ 25 , 26 ]. These reported works implied a probable reality that the catalysis activities of MXene could be originated from the 2D basal planes rather than the defects, which was significantly different from conventional nanozymes including metallic oxide [ 27 – 29 ], carbon nanomaterials [ 30 , 31 ], and 2D metallic sulfide [ 32 , 33 ]. As a result, the electrocatalytic activity of MXene was directly related to the area of flake rather than the shape or morphology. Benefiting from the unique electrocatalytic characteristics and 2D structure, MXenes were expected to provide a chance to easily control the electrocatalytic activity by simply customize the total area of flakes. In this work, the electrocatalytic activity of Ti 3 C 2 T x MXene for phenols oxidation was identified and applied to constructing a cascading catalytic amplification strategy for electrochemical biosensor to determine of BCR/ABL fusion gene, the key biomarker for clinical diagnosis of chronic myeloid leukemia (CML). Ti 3 C 2 T x MXene presented efficient and area-dependent phenol adsorption on the 2D plane, thus catalyzing the electrochemical oxidation. For biosensor application, Ti 3 C 2 T x MXene was spread on electrode and further decorated with gold nanoparticles for DNA capture probe (CP) immobilization. Besides, DNA walking machine was employed to recognize target BCR/ABL fusion gene and mediate nucleic acid amplification. As illustrated in Scheme 1 , the DNA walking machine would start DNA nicking and expose DNA fragments from the magnetic beads in the presence of BCR/ABL fusion gene. The exposed DNA fragments helped the assembly of biotin labeled DNA probe (Bio-DP) on the sensing surface according to the sandwich DNA hybridization of CP-DNA fragment-Bio-DP. Finally, streptavidin modified alkaline phosphatase (SA-ALP) further was modified onto the biosensing interface via the specific biotin-streptavidin reaction. With the addition of 1-naphthyl phosphate (1-NPP) in the electrolyte solution, 1-naphthol was produced via ALP-catalytic hydrolysis of 1-NPP and generated an amplified electrochemical signal via Ti 3 C 2 T x MXene-catalytic electrochemical oxidization. With DNA walking machine and cascading catalysis for signal amplification, the electrochemical biosensor achieved excellent sensitivity for detection of BCR/ABL fusion gene with the linear range from 0.2 fM to 20 nM and limit of detection down to 0.05 fM, which could provide a powerful bioanalysis tool for clinical diagnose of CML. Moreover, the efficient electrocatalysis activity of Ti 3 C 2 T x MXene for phenols oxidation possessed great application potential in the more fields including sewage treatment and organic synthesis and so on. 2. Experimental Section 2.1. Reagents and materials Ti 3 C 2 T x (MXene) few layer dispersion solution (Lateral size 2-5 µm), Fe 3 O 4 nanoparticles, TiO 2 nanoparticles, bulk Ti 3 AlC 2 and WS 2 nanosheets (Diameter 2-5 µm) were obtained from Jiangsu XFNANO Materials Tech. Co., Ltd. (Nanjing, China). MoS 2 nanosheets (Diameter 20-500 nm) were obtained from Nanjing JCNANO Tech. Co., Ltd. (Nanjing, China). NH 2 -Fe 3 O 4 and Nafion solutions were obtained from Aladdin Biochemical Tech. Co., Ltd. (Shanghai, China). Gold chloride (HAuCl 4 ∙4H 2 O), sodium citrate, 6-mercaptohexanol (MCH), 1-naphthyl phosphate (1-NPP), 1-naphthol, streptavidin-alkaline phosphatase (SA-ALP), 4-nitrophenol, β-estradiol and diethanolamine (DEA) were purchased from Sigma-Aldrich Chemical (St. Louis, USA). Tris(2-carboxyethyl) phosphine hydrochloride (TCEP) was purchased from Sangon Biotech. Co., Ltd. (Chongqing, China). Nt.BsmAI nicking endonuclease (Nt.BsmAI) and CutSmart buffer were provided by New England Biotech. Co., Ltd. (Beijing, China). All high-performance liquid chromatography (HPLC)-purified sequences (Table S1) in our experiments were ordered from Sangon Biotech. Co., Ltd. (Shanghai, China). Clinical serum samples were obtained from the University-Town Hospital of Chongqing Medical University (Chongqing, China). The buffers and solutions involved in this experiment were display in Supplementary Material S1. 2.2. Modification of electrode surface Prior to modification, the bare glassy carbon electrode (GCE) was polished with 300 nm and 50 nm alumina slurries to a mirror-like surface and then rinsed ultrasonically with ultrapure water, anhydrous ethanol and ultrapure water for 5 min in sequence. Then, the GCE was dried by nitrogen at room temperature. Simultaneously, Ti 3 C 2 T x MXene (0.1 mg/mL) was suspended in ultrapure water containing a 0.1% Nafion solution and sonicated for 60 min. Gold nanoparticles (AuNPs) were synthesized according to a typical method, and the detailed procedure was elaborated in Supplementary Material S3. The products were stored at 4°C protected from light for further use. Next, 10 µL of the Ti 3 C 2 T x MXene (0.10 mg/mL) suspension was dropped onto the GCE surface and allowed to dry slowly, followed by the addition of 10 µL AuNPs solution and drying at room temperature to obtain the modified electrodes (AuNPs/Ti 3 C 2 T x MXene/GCE). Afterwards, 10 µL of thiolated capture probe (CP) pretreated by TCEP was dropped onto the AuNPs/Ti 3 C 2 T x MXene/GCE surface and incubated overnight at 4°C. After being washed with washing buffer, the modified electrode was further incubated with 1.0 mM MCH for 1 h at room temperature to block the nonspecific site, obtaining the electrochemical biosensing platform (MCH/CP/AuNPs/Ti 3 C 2 T x MXene/GCE). 2.3. Preparation of DNA walking machine DNA walking machine was prepared with reference to a previous report [ 34 ]. First, 1.0 µL walker probe solution (2.0 µM) and 1.0 µL protecting probe solution (2.0 µM) were mixed and heated at 95°C for 5 min, and then naturally cooled to obtain ds-DNAs. Later, 20 µL support probe solution (2.0 µM) was sufficiently mixed with dsDNA and added to 20 µL Au@Fe 3 O 4 (the detailed procedure was described in Supplementary Material S4), stirred overnight and magnetically separated to obtain the expected DNA walking machine (DNA-Au@Fe 3 O 4 ). Finally, the well prepared DNA walking machine was further suspended in PBS and stored at 4°C for further use. 2.4. Procedure for BCR/ABL fusion gene detection First, BCR/ABL fusion gene standard sample (1.0 µL) at different concentrations, 10 U Nt.BsmAⅠ and were added in to the dispersion of DNA walking machine and kept for 2 h at 37°C. Then, supernatant solution was collected numerous after magnetic separation, which contained the produced intermediate DNAs. After that, 10 µL of the above supernatant solution and 10 µL of 2.5 µM biotinylated detection probe were dipped onto the electrode and incubated at 37°C for 1 h. Rinsing with washing buffer, the obtained electrode was treated in 10 µL of DEA buffer containing 1.25 µg/mL SA-ALP and 8 mg/mL BSA at 37°C for 30 min. Finally, the electrochemical signal was measured in the DEA buffer containing 1.0 mg/mL 1-NPP by differential pulse voltammetry (DPV) after rinsing with DEA buffer to remove the unbound SA-ALP. All parameter configurations of electrochemical measurements were shown in Supplementary Material S5. 2.5. Theoretic calculation methods The first principle calculations are performed by Vienna Ab initio Simulation Package (VASP) [ 35 ] with the projector augmented wave (PAW) method [ 36 ]. The exchange-functional is treated using the Perdew-Burke-Ernzerhof (PBE) [ 37 ] functional, in combination with the DFT-D correction [ 38 ]. The cut-off energy of the plane-wave basis is set at 500 eV. For the optimization of both geometry and lattice size, the Brillouin zone integration is performed with 2*2*1 Monkhorts-Pack k-point sampling. The self-consistent calculations apply a convergence energy threshold of 10 −5 eV. The equilibrium geometries and lattice constances are optimized with maximum stress on each atom within 0.02 eV/Å. 3. Results And Discussion 3.1. Morphological and elemental analysis of the Ti 3 C 2 T x MXene Transmission electron microscopy (TEM) was employed to study the morphology of the used Ti 3 C 2 T x MXene sample, which presented a remarkably large flake and some stacked little fragments (Fig. 1 A). Moreover, the high-angle annular dark-field (HAADF)-STEM image showed no observable spot on the flake, suggesting the uniform distribution of the elements (Fig. 1 B). As shown in Fig. 1 C- 1 E, STEM-EDS elemental mappings of C, Ti, and O presented outlines well matched with the HAADF-STEM image, which visually displayed the elemental composition of the Ti 3 C 2 T x MXene. Energy dispersive X-ray spectroscopy (EDX) was also utilized to analyze the elemental composition of the Ti 3 C 2 T x MXene (see in Supplementary Material, Fig. S1), which involved of C, O, Ti, F and Al elements. Among them, F and Al were mainly from the residual impurities and their contents were notably lower than C, O, and Ti. Atomic force microscope (AFM) was employed to further study the morphology of Ti 3 C 2 T x MXene. As shown in Fig. 1 G and 1 H, AFM image presented sheets with thickness of about 4 nm, corresponding to the thickness of 3 layers. 3.2. Electrocatalytic activity of Ti 3 C 2 T x MXene for phenolic compound oxidation The electrocatalytic activity for phenolic compound oxidation of Ti 3 C 2 T x MXene was confirmed by testing the electrocatalytic performances with different phenolic substrates, including 1-naphthol, 4-nitrophenol, and β-estradiol. As shown in Fig. 2 the modification of MXene significantly improved the oxidation currents for all the three phenolic compounds (DPV curves were seen in Supplementary Material, Fig. S2), indicating the favourable and comprehensive electrocatalytic activity of Ti 3 C 2 T x MXene for phenolic compound oxidation. Moreover, it's notable that the oxidation peaks presented distinct shifts to lower potential, revealing the electron transfer between Ti 3 C 2 T x MXene and phenolic compound. To further profile the unique electrocatalytic activity of Ti 3 C 2 T x MXene, several nanomaterials including MoS 2 nanosheets, WS 2 nanosheets, Fe 3 O 4 nanoparticles, TiO 2 nanoparticles and bulk Ti 3 AlC 2 were separately employed as constrast with 1-naphthol as substrate. It's notable that only Ti 3 C 2 T x MXene presented observable electrocatalytic activity for 1-naphthol oxidation, indicating that the unique electrocatalytic activity was the intrinsic property of Ti 3 C 2 T x MXene. To quantitatively present the electrocatalytic activity of Ti 3 C 2 T x MXene for 1-naphthol oxidation, DPV curves were measured with the addition of 1-naphthol at different concentrations. As shown in Fig. 3 A, the peak current increased with the increasing concentration of 1-naphthol at low concentrations until reaching about 90 µA. According to Faraday's laws of electrolysis and Michaelis-Menten equation, the fitting curve was achieved with hyperbola function, where the Michaelis constant was calculated to be 0.22 mM, indicating that Ti 3 C 2 T x MXene possessed strong affinity to 1-naphthol [ 39 ]. Moreover, the catalytic activity of Ti 3 C 2 T x MXene for 1-naphthol oxidation in homogeneous phase solution was explored using hydrogen peroxide as oxidant. Fig. 3 B displayed the real-time absorbance of the aqueous solutions at 387.5 nm, including 1-naphthol (curve a), mixture of 1-naphthol and hydrogen peroxide (curve b), mixture of 1-naphthol, hydrogen peroxide and Ti 3 C 2 T x MXene (curve c), respectively. It could be seen that the reaction ratio was significantly improved with the addition of Ti 3 C 2 T x MXene, indicating that Ti 3 C 2 T x MXene could efficiently catalyze the 1-naphthol oxidation by hydrogen peroxide as well [ 40 ] . To survey the quantitative relation between electrocatalytic activity and amount of Ti 3 C 2 T x MXene, Ti 3 C 2 T x MXene solutions at different concentrations were employed to modify GCE to test the electrocatalytic activities for 1-naphthol oxidation, respectively. As shown in Fig. 3 C, the Faradic current increased linearly with the increasing concentration of Ti 3 C 2 T x MXene in low concentration range and reached a constant in high concentration range, indicating that the electrocatalytic activity was in direct proportion to the total modified area of Ti 3 C 2 T x MXene flakes but not further improved by the stacking of the flakes. 3.3. Mechanism study for the catalytic activity of Ti 3 C 2 T x MXene The first principle calculations are performed to explain the catalytic activity of Ti 3 C 2 T x MXene for 1-naphthol oxidation. The free energies of 1-naphthol adsorption were first calculated to determine whether 1-naphthol could be adsorbed by Ti 3 C 2 T x MXene. The optimized geometries of 1-naphthol on and Ti 3 C 2 T x MXene were given in Fig. S3 (see in Supplementary Material), which showed 1-naphthol molecule adsorbed on the plane surface of Ti 3 C 2 T x MXene in a \"lying-down\" or \"standing-up\" manner, with adsorption energies of -1.18476 and -0.75625 eV, respectively, indicating relative strong physical adsorption. As a result, the \"lying-down\" adsorption of 1-naphthol on Ti 3 C 2 T x MXene was critical in the electrocatalytic oxidation. To further investigate the origin of 1-naphthol adsorption, the differential charge density of 1-naphthol adsorbed on Ti 3 C 2 T x MXene was calculated. As shown in Fig. 3 D, the changes in charge density caused by 1-naphthol adsorption mainly came from hydroxyl group and oxygen atoms. Moreover, changes in charge density were also found on the aromatic rings, indicating that the aromatic structure played a role in leading to the \"lying-down\" adsorption mode of 1-naphthol on Ti 3 C 2 T x MXene. 3.4. Characterization of the DNA walking machine To quantify the efficiency of DNA Walking machine, fluorescence dynamics experiments were carried out to verify it. Firstly, AuNPs and ssDNA-functionalized AuNPs were prepared according to previous literature with little adjustment [ 41 ]. In short, sodium citrate (3 mL, 1%) was added rapidly to the boiling solution of HAuCl 4 (100 mL, 1%). After the color changed from pale yellow to wine-red, the mixture was stopped heating and cooled to room temperature (RT) with continued stirring. The AuNPs had an average particle size of 13 nm and were stored at 4°C for further use. The preparation method of ssDNA (support probe and walker-protect dsDNA) -functionalized AuNPs is as follows. Firstly, the 1428 µL denature-supporting probe (2 µM) and 72 µL denature-walker-protect dsDNA (2 µM) were mixed with 1 µL acetic acid (500 mM, pH 5.2) and 0.5 µL TCEP (100 mM) at RT for 1 h, respectively. Then the mixture was added into 1 mL AuNP solution, and the resultant solution was stored in a drawer at RT for at least 16 h. After 25 µL Tris-acetate (500 mM, pH 8.2) was added to the mixture, 250 µL NaCl (1 M) was dropwise added to the mixture every three hours (30, 40, 50, 60, 70 µL were added respectively). Subsequently, the resulting mixture was stored in a drawer overnight. Lastly, the mixture was centrifuged (10 000 rpm, 10 min) to remove the excess reagents, and the red precipitate was washed and dispersed in DNA preparation solution for further use. Secondly, fluorescence kinetics curve was shown in Fig. 4 A to prove the cutting efficiency of Nt.BsmAI nicking endonuclease (Nt.BsmAI). The curve a, b and c showed corresponding changes when different concentrations of target BCR/ABL fusion gene and 10 U Nt.BsmAI were added into the ssDNA (support probe and walker-protect dsDNA) -functionalized AuNPs solution, respectively. Curve d was the blank control. The slope of the curve reflects the reaction rate of enzyme shearing, which is correlated with the concentration of target gene. As can be seen from the figure, the reaction rate is fast. At the same time, when the reaction reached 2 hours, the shearing enzyme still did not reach the maximum shearing value, which means that the shearing enzyme has not been completely reacted. Thus, the cutting efficiency of the Nt.BsmAI is excellent. In addition, the time of releasing hairpin structure DNA can also be known from the curve. The sharply rising stage in the curve mainly the enzymatic cleaving on the prehybridized Support DNA-Walker. It can be seen from the figure that it takes about 6-8 minutes to cleave and release the hairpin. The slope of tangent at t = 0s was calculated with the fitting curves indicating the cleaving rate was highly related to the concentration of substrate in Fig. 4 B. The slope of tangent at t = 0s and logarithmic value of target BCR/ABL fusion gene concentrations presents well linear dependence range from 2 pM to 2 µΜ with pearson correlation coefficient of 0.99192, which is corresponding to the kinetic characteristic of first-order reaction. Thirdly, as for the time of hairpin structure intermediate DNAs hybridized with the thiolated capture DNA on the electrode surface to form the sensing interface, we also conducted relevant optimization experiments. As shown in Fig. 4 C, the time needed was only 1h. Lastly, we have consulted relevant literature, which shows that the amount of DNA loading on 15 nm gold nanoparticles is 20-30 when the concentration of NaCl is 140mM [ 42 ]. According to the concentration and dosage of AuNPs, the amount of ssDNA fabricated on the interface of this sensor was calculated to be about 1×10 10 . 3.5. Optimization of experimental conditions To achieved optimal analytical performance of the biosensor, some critical experimental conditions were optimized, including the ratio of walker probe to support probe, the cleaving time of Nt.BsmAI nicking endonuclease, the pH of DEA buffer and the cleaving temperature of Nt.BsmAI nicking endonuclease. As shown in Fig. 5 , optimal ratio of walker probe to support probe, cleaving time, pH of DEA buffer and temperature were achieved to be 1:20, 120 min, 9.6 and 37°C, respectively. 3.6. Analytical performance of the proposed electrochemical biosensor To estimate analytical performance of the biosensor, the current responses toward BCR/ABL fusion gene at different concentrations were recorded under the optimal conditions through DPV measurements. As shown in Fig. 6 A, the detection signal increased with the increasing concentration of target BCR/ABL fusion gene. The corresponding calibration plots of the peak currents showed a strong linear relationship to the logarithm value of target BCR/ABL fusion gene concentrations range from 0.2 fM to 20 nΜ with pearson correlation coefficient of 0.99836 (Fig. 6 B). The linear regression equation was I = 1.00012 × lg ( c/ pM) + 11.23074 ( c and I stood for the concentration of target BCR/ABL fusion gene and corresponding peak current value, respectively). The limit of detection was obtained based on three times the average standard deviation corresponding to blank sample detection, which was calculated to be 0.05 fM. Comparisons of this biosensor with some reported works for BCR/ABL fusion gene detection are shown in Table S2, which highlighted the excellent sensitivity of this method in BCR/ABL fusion gene detection due to the cascading catalytic strategy and DNA walking machine for signal amplification. Moreover, the specificity of the biosensor was evaluated by using 3 different DNA oligonucleotides as references, including a single-base-mismatched strand (B1), a two-base-mismatched strand (B2) and a noncomplementary strand (B3), all at concentrations of 20 fM. As depicted in Fig. 6 C, the response signals of the single-base-mismatched strand and two-base-mismatched strand were much lower than the response signal of the target, revealing the good capacity of the biosensor to distinguish base-mismatch. The response signal of noncomplementary sequences were approximate to the blank solution, indicating that the biosensor presented good selectivity for DNA detection. To evaluate the stability of the proposed biosensor, the modified electrodes were stored at 4°C before use. As presented in Fig. 6 D, there were no obvious differences during the first 5 days of storage, and the current changes were less than 1.58%. After 20 days of storage, the designed biosensor retained 89.40% of its initial current response, indicating that the proposed biosensor offers satisfactory stability for target BCR/ABL detection. 3.7. Detection of BCR/ABL fusion gene in human serum samples To further validate the applicability of the biosensor to complex biological matrix in clinical application, different concentrations of target BCR/ABL fusion gene were added to 10-fold-diluted clinical serum samples and tested with the proposed biosensor. The detection results of BCR/ABL fusion gene in human serum samples are summarized in Table S3. Satisfactory recovery values were obtained ranging from 93.60–110.42% with relative standard deviations (RSD) between 0.27% and 0.64%. In addition, we extracted RNA from clinical serum of BCR/ABL positive patients using spin columns CB3 according to the manufacture's protocol and tested with the proposed biosensor. The detection concentrations of BCR/ABL were compared with clinical results (by reverse transcription PCR), which were summarized in Table S4. It could be seen that the proposed biosensor achieved results well matched with the clinical assay, manifesting the application potential of the proposed biosensor in clinical diagnosis. 4. Conclusions In summary, this work reported the unique catalytic activity of Ti 3 C 2 T x MXene nanozyme for phenols oxidation and the application to electrochemical biosensor for BCR/ABL fusion gene detection. The catalytic activity originated from the adsorption capacity of the Ti 3 C 2 T x MXene plane surface towards 1-naphthol. Theoretic calculation revealed the mechanism that relatively strong interaction was existed between the surface oxygen atoms of Ti 3 C 2 T x MXene and the phenolic hydroxyl groups. Moreover, the catalytic activity was strictly proportional to the cover area of the MXene fakes but independent to the number of stacking layers, which was quite different from traditional nanozymes and significant benefited the quality control of biosensor. Ultrasensitive detection was achieved with the proposed biosensor, which provided a promising analytical tool for the clinical diagnosis of CML. Declarations Acknowledgments This research was supported by the National Natural Science Foundation of China (Grant Nos. 31071093, 31170129, and 31200064), China Postdoctoral Science Foundation (2020M683259) and the Science and Technology Planning Project of Yuzhong District of Chongqing city, China (No. 20140119). Authors’ contributions Rongjun Yu: Conceptualization, Methodology, Software, Investigation, Data Curation, Writing-Original Draft. Jian Xue: Software, Investigation, Project Administration. Yang Wang: Formal Analysis, Software. Jingfu Qiu: Supervision, Methodology, Data Curation, Administration. Xinyi Huang: Methodology, Software, Investigation, Project Administration. Anyi Chen: Writing-Review & Editing. Jianjiang Xue: Funding Acquisition, Supervision, Administration, Data Curation. Availability of data and materials All data generated or analyzed during this study are included in this article and the Additional Information. Consent for publication All authors of this study agreed to publish. Competing interests The authors declare no competing financial interests. 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Zhao X, Xu H, Hui Z, Sun Y, Yu C, Xue J, Zhou R, Wang L, Dai H, Zhao Y, Yang J, Zhou J, Chen Q, Sun G, Huang W Electrostatically Assembling 2D Nanosheets of MXene and MOF-Derivatives into 3D Hollow Frameworks for Enhanced Lithium Storage. Small. 2019; 15: e1904255. Lang Z, Zhuang Z, Li S, Xia L, Zhao Y, Zhao Y, Han C, Zhou L. MXene Surface Terminations Enable Strong Metal–Support Interactions for Efficient Methanol Oxidation on Palladium. ACS Appl Mater Interfaces. 2020;12:2400–6. Wang H, Lee J. Recent advances in structural engineering of MXene electrocatalysts. J MaterChem A. 2020;8:10604–24. Gao L, Fan K, Yan X. Iron Oxide Nanozyme: A Multifunctional Enzyme Mimetic for Biomedical Applications. Theranostics. 2017;7:3207–27. Pratsinis A, Kelesidis GA, Zuercher S, Krumeich F, Bolisetty S, Mezzenga R, Leroux J-C. ,Sotiriou G A. Enzyme-Mimetic Antioxidant Luminescent Nanoparticles for Highly Sensitive Hydrogen Peroxide Biosensing. ACS Nano. 2017;11:12210–8. Zhu Y, Zhang Z, Song X, Bu Y. A facile strategy for synthesis of porous Cu2O nanospheres and application as nanozymes in colorimetric biosensing. J Mater Chem B. 2021;9:3533–43. Fan K, Xi J, Fan L, Wang P, Zhu C, Tang Y, Xu X, Liang M, Jiang B, Yan X. Gao L. In vivo guiding nitrogen-doped carbon nanozyme for tumor catalytic therapy. Nat Commun. 2018;9:1440. Zhang P, Sun D, Cho A, Weon S, Lee S, Lee J, Han JW, Kim D-P. Choi W. Modified carbon nitride nanozyme as bifunctional glucose oxidase-peroxidase for metal-free bioinspired cascade photocatalysis. Nat Commun. 2019;10:940. Chen T, Zou H, Wu X, Liu C, Situ B, Zheng L. Yang G. Nanozymatic Antioxidant System Based on MoS2 Nanosheets. ACS Appl Mater Interfaces. 2018;10:12453–62. Zhu J, Peng X, Nie W, Wang Y, Gao J, Wen W, Selvaraj JN, Zhang X, Wang S. Hollow copper sulfide nanocubes as multifunctional nanozymes for colorimetric detection of dopamine and electrochemical detection of glucose. Biosens Bioelectron. 2019;141:111450. Xu Z, Liao L, Chai Y, Wang H, Yuan R. Ultrasensitive Electrochemiluminescence Biosensor for MicroRNA Detection by 3D DNA Walking Machine Based Target Conversion and Distance-Controllable Signal Quenching and Enhancing. Anal Chem. 2017;89:8282–7. Kresse G Furthmüller. J. Efficiency of ab-initio total energy calculations for metals and semiconductors using a plane-wave basis set. Comput Mater Sci. 1996;6:15–50. Joubert D, Kresse G. From ultrasoft pseudopotentials to the projector augmented-wave method. Phys Rev B. 1999;59:1758–75. Burke K, Ernzerhof M, Perdew JP Generalized Gradient Approximation Made Simple [Phys. Rev. Lett. 77, 3865 (1996)]. Phys Rev Lett 1997; 78: 1396-. Grimme S, Antony J, Ehrlich S, Krieg H. A consistent and accurate ab initio parametrization of density functional dispersion correction (DFT-D) for the 94 elements H-Pu. J Chem Phys. 2010;132:154104. Wang J, Huang R, Qi W, Su R, Binks BP, He Z. Construction of a bioinspired laccase-mimicking nanozyme for the degradation and detection of phenolic pollutants. Appl Catal B. 2019;254:452–62. Jiang B, Duan D, Gao L, Zhou M, Fan K, Tang Y, Xi J, Bi Y, Tong Z, Gao GF, Xie N, Tang A, Nie G, Liang M. Yan X. Standardized assays for determining the catalytic activity and kinetics of peroxidase-like nanozymes. Nat Protoc. 2018;13:1506–20. Juewen L, Yi L. Preparation of aptamer-linked gold nanoparticle purple aggregates for colorimetric sensing of analytes. Nat Protoc. 2006;1:246–52. Sarah J, Hurst AKR, Lytton-J.,Chad AM. Maximizing DNA Loading on a Range of Gold Nanoparticle Sizes. Anal Chem. 2006;78:8313–8. scheme Scheme 1 is available in the Supplemental Files section Supplementary Files GraphicalAbstract.docx Scheme 1 Schematic illustration for the working principle of the electrochemical biosensor. RevisedSupplementaryMaterial.docx Cite Share Download PDF Status: Published Journal Publication published 09 Mar, 2022 Read the published version in Journal of Nanobiotechnology → Version 2 posted Editorial decision: Accept 20 Feb, 2022 Reviews received at journal 31 Jan, 2022 Reviewers invited by journal 30 Jan, 2022 Editor invited by journal 20 Jan, 2022 Editor assigned by journal 20 Jan, 2022 First submitted to journal 16 Jan, 2022 You are reading this latest preprint version Show more versions Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {\"props\":{\"pageProps\":{\"initialData\":{\"identity\":\"rs-965061\",\"acceptedTermsAndConditions\":true,\"allowDirectSubmit\":false,\"archivedVersions\":[{\"code\":1,\"date\":\"2021-10-19 19:24:55\",\"editorialEvents\":[{\"type\":\"communityComments\",\"content\":0}],\"status\":\"published\",\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"researchsquare\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":true,\"externalIdentity\":\"\",\"sideBox\":\"\",\"snPcode\":\"\",\"submissionUrl\":\"/submission\",\"title\":\"Research 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University\",\"correspondingAuthor\":false,\"submittingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Jian\",\"middleName\":\"\",\"lastName\":\"Xue\",\"suffix\":\"\"},{\"id\":83777987,\"identity\":\"fa13c5c0-72d3-4b01-8650-5b0d14518542\",\"order_by\":2,\"name\":\"Yang Wang\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"University-Town Hospital of Chongqing Medical University\",\"correspondingAuthor\":false,\"submittingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Yang\",\"middleName\":\"\",\"lastName\":\"Wang\",\"suffix\":\"\"},{\"id\":83777988,\"identity\":\"12e60d41-3ac6-46b8-a453-cd2452012503\",\"order_by\":3,\"name\":\"Jingfu Qiu\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Chongqing Medical University\",\"correspondingAuthor\":false,\"submittingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Jingfu\",\"middleName\":\"\",\"lastName\":\"Qiu\",\"suffix\":\"\"},{\"id\":83777989,\"identity\":\"5b7c23c0-169d-4683-bb48-7c76f09a100b\",\"order_by\":4,\"name\":\"Xinyi Huang\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"First Affiliated Hospital of Guangxi University of Chinese Medicine.\",\"correspondingAuthor\":false,\"submittingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Xinyi\",\"middleName\":\"\",\"lastName\":\"Huang\",\"suffix\":\"\"},{\"id\":83777990,\"identity\":\"e1c7d370-4650-4b0a-a267-bf71fbcd8f55\",\"order_by\":5,\"name\":\"Anyi Chen\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Chongqing Medical University\",\"correspondingAuthor\":false,\"submittingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Anyi\",\"middleName\":\"\",\"lastName\":\"Chen\",\"suffix\":\"\"},{\"id\":83777991,\"identity\":\"f2cfb152-4c48-4db2-a933-d0e1c2d653f6\",\"order_by\":6,\"name\":\"jianjiang xue\",\"email\":\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA4klEQVRIiWNgGAWjYBACxmYgIWEA5X0wsLEjTQvjjIK0ZNKsZOb5cIixgaCqduZnDywK7PLkI3IMH9sYHGBmYD98dAN+h7GZG0gYJBcbnjljbJxjcIePgSct7QZ+LQxmEhIGzIkb23vMpHMMnjEzSPCYEdDC/g2opT5xYzOP+W8Lg8OMDYS18IBsOZw4n73HjJmBSC1lQC3HEzfwHCuW7DFIS2Yj5BfD/uPbpCX+VCfOn5G88cOPPzZ2/OyHj+HX0gAMaAkgw+AAVIQNn3IQkAc57gOI0UBI6SgYBaNgFIxYAACNa0QtlVlItQAAAABJRU5ErkJggg==\",\"orcid\":\"\",\"institution\":\"University-Town Hospital of Chongqing Medical University\",\"correspondingAuthor\":true,\"submittingAuthor\":false,\"prefix\":\"\",\"firstName\":\"jianjiang\",\"middleName\":\"\",\"lastName\":\"xue\",\"suffix\":\"\"}],\"badges\":[],\"createdAt\":\"2021-10-12 10:33:57\",\"currentVersionCode\":2,\"declarations\":\"\",\"doi\":\"10.21203/rs.3.rs-965061/v2\",\"doiUrl\":\"https://doi.org/10.21203/rs.3.rs-965061/v2\",\"draftVersion\":[],\"editorialEvents\":[{\"content\":\"https://doi.org/10.1186/s12951-022-01317-9\",\"type\":\"published\",\"date\":\"2022-03-09T12:00:34+00:00\"}],\"editorialNote\":\"\",\"failedWorkflow\":false,\"files\":[{\"id\":18257985,\"identity\":\"19effa5c-2d9f-443c-a389-c384abe039ff\",\"added_by\":\"auto\",\"created_at\":\"2022-02-15 23:00:40\",\"extension\":\"png\",\"order_by\":1,\"title\":\"Figure 1\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":1464182,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003e(A) TEM image, (B) HAADF-STEM image and (C-F) STEM-EDS elemental mappings of the Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene flakes. (G) AFM image of the Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene flakes and (H) the height profile along the white line.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"1.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-965061/v2/6339186427b4c78a890ca9e7.png\"},{\"id\":18257980,\"identity\":\"f0b083f4-3cd5-4aa6-8273-0733b1c6d648\",\"added_by\":\"auto\",\"created_at\":\"2022-02-15 23:00:40\",\"extension\":\"png\",\"order_by\":2,\"title\":\"Figure 2\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":134504,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003e(A) Oxidation currents of different phenolic samples measured with bare GCE and Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene modified electrode, respectively; (B) Oxidation current of 1-naphthol measured with different materials modified electrodes.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"2.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-965061/v2/2acc8a539b0d1cf56e752bdf.png\"},{\"id\":18258062,\"identity\":\"e8fc4b3a-95ba-4750-86ba-1ac1cf2a0ce0\",\"added_by\":\"auto\",\"created_at\":\"2022-02-15 23:03:41\",\"extension\":\"png\",\"order_by\":3,\"title\":\"Figure 3\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":551252,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003e(A) Oxidation currents of 1-naphthol at different concentrations measured with Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene modified electrode; (B) Absorbancy at 387.5 nm of 1-naphthol solution (curve a), mixture of 1-naphthol and hydrogen peroxide (curve b), mixture of 1-naphthol, hydrogen peroxide and Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene (curve c), respectively. (C) The Faradic currents measured by electrode modified with Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene at different concentrations in solution containing 7.0 mM 1-naphthol. (D) Charge density difference of the \\\"lying-down\\\" adsorption mode. Yellow and blue represent charge accumulation and depletion (isovalue: 0.005 au).\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"3.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-965061/v2/b8628e351561a7036dd9c19c.png\"},{\"id\":18257975,\"identity\":\"d6b4438d-ef94-4837-bb3a-903d7b296786\",\"added_by\":\"auto\",\"created_at\":\"2022-02-15 23:00:40\",\"extension\":\"png\",\"order_by\":4,\"title\":\"Figure 4\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":144931,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003e(A) Fluorescence kinetics curves of the nanomachine with addition of 10 U Nt.BsmAI and target BCR/ABL fusion gene at different concentrations (from a to d: 2 μM, 2 nM, 2 pM and blank, separately). (B) The relationship between the slope of tangent at t = 0s and logarithmic value of target BCR/ABL fusion gene concentrations (range of concentration: 2 pM, 2 nM, and 2 uM separately). (C) Time optimization of intermediate DNAs hybridizing with the capture DNA.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"4.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-965061/v2/0eb1caa4d11242d992d8dd92.png\"},{\"id\":18257983,\"identity\":\"9d4321f9-947e-45c3-9cb1-161b48ef8b2d\",\"added_by\":\"auto\",\"created_at\":\"2022-02-15 23:00:40\",\"extension\":\"png\",\"order_by\":5,\"title\":\"Figure 5\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":97844,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eOptimization of experimental conditions: (A) ratio of walker to support DNA, (B) cleaving time of Nt.BsmAI nicking endonuclease, (C) pH of DEA buffer, (D)\\u0026nbsp;cleaving temperature of Nt.BsmAI nicking endonuclease. The error bars represent the standard deviation of three parallel measurements.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"5.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-965061/v2/4ada413f979db1cd499fbfd0.png\"},{\"id\":18257987,\"identity\":\"1fa61fe5-915c-4015-89cd-4af565110989\",\"added_by\":\"auto\",\"created_at\":\"2022-02-15 23:00:40\",\"extension\":\"png\",\"order_by\":6,\"title\":\"Figure 6\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":145977,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eEvaluation of the sensitivity and specificity of the biosensor: (A) DPV curves response of the electrochemical biosensor upon the increase in target BCR/ABL fusion gene concentration (from bottom to top: 0 fM, 0.2 fM, 2 fM, 20 fM, 200 fM, 2 pM, 20 pM, 200 pM, and 2 nM, 20 nM, separately) and (B) the corresponding linear relationship between DPV signal and logarithmic value of target BCR/ABL fusion gene concentrations (range of concentration: 20 aM, 0.2 fM, 2 fM, 20 fM, 200 fM, 2 pM, 20 pM, 200 pM, and 2 nM, 20 nM, 200 nM, 2 uM separately). (C)\\u003cstrong\\u003e \\u003c/strong\\u003eDPV\\u003cstrong\\u003e \\u003c/strong\\u003eresponses of the electrochemical biosensor to different oligonucleotides (20 fM): (a) BCR/ABL fusion gene (target), (b) single-base-mismatched strand (B1), (c) two-base-mismatched strand (B2), (d) noncomplementary strand (B3), and (e) blank. (D) Stability of the proposed biosensor. The error bars represent the standard deviation of three parallel measurements.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"6.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-965061/v2/b9fa50faf29486c122e8ba8e.png\"},{\"id\":19023151,\"identity\":\"dfcfda22-685e-4048-9090-0b6c85023c48\",\"added_by\":\"auto\",\"created_at\":\"2022-03-09 12:00:37\",\"extension\":\"pdf\",\"order_by\":0,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"manuscript-pdf\",\"size\":2882558,\"visible\":true,\"origin\":\"\",\"legend\":\"\",\"description\":\"\",\"filename\":\"manuscript.pdf\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-965061/v2/7dd59f50-6a5b-47e1-b07e-1048b9634aa0.pdf\"},{\"id\":18257984,\"identity\":\"dfb3a6d6-0920-41f7-bda7-57c3e0cc9865\",\"added_by\":\"auto\",\"created_at\":\"2022-02-15 23:00:40\",\"extension\":\"docx\",\"order_by\":1,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"supplement\",\"size\":565916,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003e\\u003cstrong\\u003eScheme 1\\u003c/strong\\u003e Schematic illustration for the working principle of the electrochemical biosensor.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"GraphicalAbstract.docx\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-965061/v2/e6809faf298409ca49e3ac92.docx\"},{\"id\":18257981,\"identity\":\"7124df50-43b4-42c8-8457-58c24f69ce8d\",\"added_by\":\"auto\",\"created_at\":\"2022-02-15 23:00:40\",\"extension\":\"docx\",\"order_by\":2,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"supplement\",\"size\":3247687,\"visible\":true,\"origin\":\"\",\"legend\":\"\",\"description\":\"\",\"filename\":\"RevisedSupplementaryMaterial.docx\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-965061/v2/8d055d64469d5a5c1d059bfd.docx\"}],\"financialInterests\":\"\",\"formattedTitle\":\"Novel Ti3C2Tx MXene Nanozyme with Manageable Catalytic Activity and Application to Electrochemical Biosensor\",\"fulltext\":[{\"header\":\"1. Introduction\",\"content\":\"\\u003cp\\u003eElectrochemical biosensor has become one of the most predominant analysis tools in clinical diagnosis due to the outstanding merits of high sensitivity and selectivity, rapid response, low cost, simple instrumentation, easy miniaturization and good quantitative ability [\\u003cspan additionalcitationids=\\\"CR2 CR3\\\" citationid=\\\"CR1\\\" class=\\\"CitationRef\\\"\\u003e1\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR4\\\" class=\\\"CitationRef\\\"\\u003e4\\u003c/span\\u003e]. Enzymatic electrocatalysis is a widely used technique in electrochemical biosensor, which improves the analytical sensitivity by promoting the electrochemical redox reactions with the help of enzymes [\\u003cspan additionalcitationids=\\\"CR6\\\" citationid=\\\"CR5\\\" class=\\\"CitationRef\\\"\\u003e5\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR7\\\" class=\\\"CitationRef\\\"\\u003e7\\u003c/span\\u003e]. Nevertheless, natural enzymes are general cost to manufacture and store, unstable to transfer or modify, and sensitive to harsh physiochemical conditions. Nanozymes, nanomaterials with enzyme-like catalytic activities, well overcome the shortages of natural enzymes [\\u003cspan citationid=\\\"CR8\\\" class=\\\"CitationRef\\\"\\u003e8\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR9\\\" class=\\\"CitationRef\\\"\\u003e9\\u003c/span\\u003e]. Therefore, nanozymes are attracting increasing attention in bioanalysis to substitute conventional natural enzyme [\\u003cspan citationid=\\\"CR10\\\" class=\\\"CitationRef\\\"\\u003e10\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR11\\\" class=\\\"CitationRef\\\"\\u003e11\\u003c/span\\u003e]. However, the catalytic activities of conventional nanozymes strongly rely on the defect dependent active centers, such as surface dangling bonds or heterogeneous doping domains, which is distributed unevenly on the surface [\\u003cspan citationid=\\\"CR12\\\" class=\\\"CitationRef\\\"\\u003e12\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR13\\\" class=\\\"CitationRef\\\"\\u003e13\\u003c/span\\u003e]. As a consequence, the homogeneity of particle is a very important parameter of nanozymes, which brings great challenge to prepare high-quality product and applied to precise quantitative analysis [\\u003cspan citationid=\\\"CR10\\\" class=\\\"CitationRef\\\"\\u003e10\\u003c/span\\u003e]. From this prospective, developing nanomaterials with enzyme-like catalytic activities independent to the morphology or crystal defect is of great significance as this kind of materials might be more easily controlled to obtain uniform catalysis activity.\\u003c/p\\u003e \\u003cp\\u003eMXene is termed for a series of two-dimension (2D) transition metal carbides, nitrides, and carbonitrides [\\u003cspan citationid=\\\"CR14\\\" class=\\\"CitationRef\\\"\\u003e14\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR15\\\" class=\\\"CitationRef\\\"\\u003e15\\u003c/span\\u003e]. Recent years, because typical MXene possess biocompatibility, large specific surface area, rich surface chemistry, tunable lateral size, good electrical conductivity, and mechanical robustness, allowing efficient and selective interaction with target species. MXene-based materials have been utilized as electrocatalysts for detecting small molecules, pharmaceutical drugs, environmental pollutants, and biomarkers. These characteristics render MXene an ideal platform as sensing materials for electrochemical application [\\u003cspan citationid=\\\"CR16\\\" class=\\\"CitationRef\\\"\\u003e16\\u003c/span\\u003e]. The electrocatalytic activity MXene have attracted enormous research interest in diverse fields, such as hydrogen evolution [\\u003cspan citationid=\\\"CR17\\\" class=\\\"CitationRef\\\"\\u003e17\\u003c/span\\u003e], oxygen evolution [\\u003cspan citationid=\\\"CR18\\\" class=\\\"CitationRef\\\"\\u003e18\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR19\\\" class=\\\"CitationRef\\\"\\u003e19\\u003c/span\\u003e], N\\u003csub\\u003e2\\u003c/sub\\u003e-to-NH\\u003csub\\u003e3\\u003c/sub\\u003e conversion [\\u003cspan citationid=\\\"CR20\\\" class=\\\"CitationRef\\\"\\u003e20\\u003c/span\\u003e], fuel cell [\\u003cspan citationid=\\\"CR21\\\" class=\\\"CitationRef\\\"\\u003e21\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR22\\\" class=\\\"CitationRef\\\"\\u003e22\\u003c/span\\u003e], energy storage [\\u003cspan citationid=\\\"CR23\\\" class=\\\"CitationRef\\\"\\u003e23\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR24\\\" class=\\\"CitationRef\\\"\\u003e24\\u003c/span\\u003e] and carbon dioxide reduction [\\u003cspan citationid=\\\"CR25\\\" class=\\\"CitationRef\\\"\\u003e25\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR26\\\" class=\\\"CitationRef\\\"\\u003e26\\u003c/span\\u003e]. These reported works implied a probable reality that the catalysis activities of MXene could be originated from the 2D basal planes rather than the defects, which was significantly different from conventional nanozymes including metallic oxide [\\u003cspan additionalcitationids=\\\"CR28\\\" citationid=\\\"CR27\\\" class=\\\"CitationRef\\\"\\u003e27\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR29\\\" class=\\\"CitationRef\\\"\\u003e29\\u003c/span\\u003e], carbon nanomaterials [\\u003cspan citationid=\\\"CR30\\\" class=\\\"CitationRef\\\"\\u003e30\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR31\\\" class=\\\"CitationRef\\\"\\u003e31\\u003c/span\\u003e], and 2D metallic sulfide [\\u003cspan citationid=\\\"CR32\\\" class=\\\"CitationRef\\\"\\u003e32\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR33\\\" class=\\\"CitationRef\\\"\\u003e33\\u003c/span\\u003e]. As a result, the electrocatalytic activity of MXene was directly related to the area of flake rather than the shape or morphology. Benefiting from the unique electrocatalytic characteristics and 2D structure, MXenes were expected to provide a chance to easily control the electrocatalytic activity by simply customize the total area of flakes.\\u003c/p\\u003e \\u003cp\\u003eIn this work, the electrocatalytic activity of Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene for phenols oxidation was identified and applied to constructing a cascading catalytic amplification strategy for electrochemical biosensor to determine of BCR/ABL fusion gene, the key biomarker for clinical diagnosis of chronic myeloid leukemia (CML). Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene presented efficient and area-dependent phenol adsorption on the 2D plane, thus catalyzing the electrochemical oxidation. For biosensor application, Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene was spread on electrode and further decorated with gold nanoparticles for DNA capture probe (CP) immobilization. Besides, DNA walking machine was employed to recognize target BCR/ABL fusion gene and mediate nucleic acid amplification. As illustrated in Scheme \\u003cspan refid=\\\"Sch1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003e, the DNA walking machine would start DNA nicking and expose DNA fragments from the magnetic beads in the presence of BCR/ABL fusion gene. The exposed DNA fragments helped the assembly of biotin labeled DNA probe (Bio-DP) on the sensing surface according to the sandwich DNA hybridization of CP-DNA fragment-Bio-DP. Finally, streptavidin modified alkaline phosphatase (SA-ALP) further was modified onto the biosensing interface \\u003cem\\u003evia\\u003c/em\\u003e the specific biotin-streptavidin reaction. With the addition of 1-naphthyl phosphate (1-NPP) in the electrolyte solution, 1-naphthol was produced \\u003cem\\u003evia\\u003c/em\\u003e ALP-catalytic hydrolysis of 1-NPP and generated an amplified electrochemical signal \\u003cem\\u003evia\\u003c/em\\u003e Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene-catalytic electrochemical oxidization. With DNA walking machine and cascading catalysis for signal amplification, the electrochemical biosensor achieved excellent sensitivity for detection of BCR/ABL fusion gene with the linear range from 0.2 fM to 20 nM and limit of detection down to 0.05 fM, which could provide a powerful bioanalysis tool for clinical diagnose of CML. Moreover, the efficient electrocatalysis activity of Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene for phenols oxidation possessed great application potential in the more fields including sewage treatment and organic synthesis and so on.\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e\"},{\"header\":\"2. Experimental Section\",\"content\":\"\\u003cdiv id=\\\"Sec3\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.1. Reagents and materials\\u003c/h2\\u003e \\u003cp\\u003eTi\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e (MXene) few layer dispersion solution (Lateral size 2-5 \\u0026micro;m), Fe\\u003csub\\u003e3\\u003c/sub\\u003eO\\u003csub\\u003e4\\u003c/sub\\u003e nanoparticles, TiO\\u003csub\\u003e2\\u003c/sub\\u003e nanoparticles, bulk Ti\\u003csub\\u003e3\\u003c/sub\\u003eAlC\\u003csub\\u003e2\\u003c/sub\\u003e and WS\\u003csub\\u003e2\\u003c/sub\\u003e nanosheets (Diameter 2-5 \\u0026micro;m) were obtained from Jiangsu XFNANO Materials Tech. Co., Ltd. (Nanjing, China). MoS\\u003csub\\u003e2\\u003c/sub\\u003e nanosheets (Diameter 20-500 nm) were obtained from Nanjing JCNANO Tech. Co., Ltd. (Nanjing, China). NH\\u003csub\\u003e2\\u003c/sub\\u003e-Fe\\u003csub\\u003e3\\u003c/sub\\u003eO\\u003csub\\u003e4\\u003c/sub\\u003e and Nafion solutions were obtained from Aladdin Biochemical Tech. Co., Ltd. (Shanghai, China). Gold chloride (HAuCl\\u003csub\\u003e4\\u003c/sub\\u003e∙4H\\u003csub\\u003e2\\u003c/sub\\u003eO), sodium citrate, 6-mercaptohexanol (MCH), 1-naphthyl phosphate (1-NPP), 1-naphthol, streptavidin-alkaline phosphatase (SA-ALP), 4-nitrophenol, β-estradiol and diethanolamine (DEA) were purchased from Sigma-Aldrich Chemical (St. Louis, USA). Tris(2-carboxyethyl) phosphine hydrochloride (TCEP) was purchased from Sangon Biotech. Co., Ltd. (Chongqing, China). Nt.BsmAI nicking endonuclease (Nt.BsmAI) and CutSmart buffer were provided by New England Biotech. Co., Ltd. (Beijing, China). All high-performance liquid chromatography (HPLC)-purified sequences (Table S1) in our experiments were ordered from Sangon Biotech. Co., Ltd. (Shanghai, China). Clinical serum samples were obtained from the University-Town Hospital of Chongqing Medical University (Chongqing, China). The buffers and solutions involved in this experiment were display in Supplementary Material S1.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec4\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.2. Modification of electrode surface\\u003c/h2\\u003e \\u003cp\\u003ePrior to modification, the bare glassy carbon electrode (GCE) was polished with 300 nm and 50 nm alumina slurries to a mirror-like surface and then rinsed ultrasonically with ultrapure water, anhydrous ethanol and ultrapure water for 5 min in sequence. Then, the GCE was dried by nitrogen at room temperature. Simultaneously, Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene (0.1 mg/mL) was suspended in ultrapure water containing a 0.1% Nafion solution and sonicated for 60 min. Gold nanoparticles (AuNPs) were synthesized according to a typical method, and the detailed procedure was elaborated in Supplementary Material S3. The products were stored at 4\\u0026deg;C protected from light for further use.\\u003c/p\\u003e \\u003cp\\u003eNext, 10 \\u0026micro;L of the Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene (0.10 mg/mL) suspension was dropped onto the GCE surface and allowed to dry slowly, followed by the addition of 10 \\u0026micro;L AuNPs solution and drying at room temperature to obtain the modified electrodes (AuNPs/Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene/GCE). Afterwards, 10 \\u0026micro;L of thiolated capture probe (CP) pretreated by TCEP was dropped onto the AuNPs/Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene/GCE surface and incubated overnight at 4\\u0026deg;C. After being washed with washing buffer, the modified electrode was further incubated with 1.0 mM MCH for 1 h at room temperature to block the nonspecific site, obtaining the electrochemical biosensing platform (MCH/CP/AuNPs/Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene/GCE).\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec5\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.3. Preparation of DNA walking machine\\u003c/h2\\u003e \\u003cp\\u003eDNA walking machine was prepared with reference to a previous report [\\u003cspan citationid=\\\"CR34\\\" class=\\\"CitationRef\\\"\\u003e34\\u003c/span\\u003e]. First, 1.0 \\u0026micro;L walker probe solution (2.0 \\u0026micro;M) and 1.0 \\u0026micro;L protecting probe solution (2.0 \\u0026micro;M) were mixed and heated at 95\\u0026deg;C for 5 min, and then naturally cooled to obtain ds-DNAs. Later, 20 \\u0026micro;L support probe solution (2.0 \\u0026micro;M) was sufficiently mixed with dsDNA and added to 20 \\u0026micro;L Au@Fe\\u003csub\\u003e3\\u003c/sub\\u003eO\\u003csub\\u003e4\\u003c/sub\\u003e (the detailed procedure was described in Supplementary Material S4), stirred overnight and magnetically separated to obtain the expected DNA walking machine (DNA-Au@Fe\\u003csub\\u003e3\\u003c/sub\\u003eO\\u003csub\\u003e4\\u003c/sub\\u003e). Finally, the well prepared DNA walking machine was further suspended in PBS and stored at 4\\u0026deg;C for further use.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec6\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.4. Procedure for BCR/ABL fusion gene detection\\u003c/h2\\u003e \\u003cp\\u003eFirst, BCR/ABL fusion gene standard sample (1.0 \\u0026micro;L) at different concentrations, 10 U Nt.BsmAⅠ and were added in to the dispersion of DNA walking machine and kept for 2 h at 37\\u0026deg;C. Then, supernatant solution was collected numerous after magnetic separation, which contained the produced intermediate DNAs. After that, 10 \\u0026micro;L of the above supernatant solution and 10 \\u0026micro;L of 2.5 \\u0026micro;M biotinylated detection probe were dipped onto the electrode and incubated at 37\\u0026deg;C for 1 h. Rinsing with washing buffer, the obtained electrode was treated in 10 \\u0026micro;L of DEA buffer containing 1.25 \\u0026micro;g/mL SA-ALP and 8 mg/mL BSA at 37\\u0026deg;C for 30 min. Finally, the electrochemical signal was measured in the DEA buffer containing 1.0 mg/mL 1-NPP by differential pulse voltammetry (DPV) after rinsing with DEA buffer to remove the unbound SA-ALP. All parameter configurations of electrochemical measurements were shown in Supplementary Material S5.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec7\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.5. Theoretic calculation methods\\u003c/h2\\u003e \\u003cp\\u003eThe first principle calculations are performed by Vienna Ab initio Simulation Package (VASP) [\\u003cspan citationid=\\\"CR35\\\" class=\\\"CitationRef\\\"\\u003e35\\u003c/span\\u003e] with the projector augmented wave (PAW) method [\\u003cspan citationid=\\\"CR36\\\" class=\\\"CitationRef\\\"\\u003e36\\u003c/span\\u003e]. The exchange-functional is treated using the Perdew-Burke-Ernzerhof (PBE) [\\u003cspan citationid=\\\"CR37\\\" class=\\\"CitationRef\\\"\\u003e37\\u003c/span\\u003e] functional, in combination with the DFT-D correction [\\u003cspan citationid=\\\"CR38\\\" class=\\\"CitationRef\\\"\\u003e38\\u003c/span\\u003e]. The cut-off energy of the plane-wave basis is set at 500 eV. For the optimization of both geometry and lattice size, the Brillouin zone integration is performed with 2*2*1 Monkhorts-Pack k-point sampling. The self-consistent calculations apply a convergence energy threshold of 10\\u003csup\\u003e\\u0026minus;5\\u003c/sup\\u003e eV. The equilibrium geometries and lattice constances are optimized with maximum stress on each atom within 0.02 eV/\\u0026Aring;.\\u003c/p\\u003e \\u003c/div\\u003e\"},{\"header\":\"3. Results And Discussion\",\"content\":\"\\u003cdiv id=\\\"Sec9\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e3.1. Morphological and elemental analysis of the Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene\\u003c/h2\\u003e \\u003cp\\u003eTransmission electron microscopy (TEM) was employed to study the morphology of the used Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene sample, which presented a remarkably large flake and some stacked little fragments (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003eA). Moreover, the high-angle annular dark-field (HAADF)-STEM image showed no observable spot on the flake, suggesting the uniform distribution of the elements (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003eB). As shown in Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003eC-\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003eE, STEM-EDS elemental mappings of C, Ti, and O presented outlines well matched with the HAADF-STEM image, which visually displayed the elemental composition of the Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene.\\u003c/p\\u003e \\u003cp\\u003eEnergy dispersive X-ray spectroscopy (EDX) was also utilized to analyze the elemental composition of the Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene (see in Supplementary Material, Fig. S1), which involved of C, O, Ti, F and Al elements. Among them, F and Al were mainly from the residual impurities and their contents were notably lower than C, O, and Ti.\\u003c/p\\u003e \\u003cp\\u003eAtomic force microscope (AFM) was employed to further study the morphology of Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene. As shown in Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003eG and \\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003eH, AFM image presented sheets with thickness of about 4 nm, corresponding to the thickness of 3 layers.\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec10\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e3.2. Electrocatalytic activity of Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene for phenolic compound oxidation\\u003c/h2\\u003e \\u003cp\\u003eThe electrocatalytic activity for phenolic compound oxidation of Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene was confirmed by testing the electrocatalytic performances with different phenolic substrates, including 1-naphthol, 4-nitrophenol, and β-estradiol. As shown in Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003e the modification of MXene significantly improved the oxidation currents for all the three phenolic compounds (DPV curves were seen in Supplementary Material, Fig. S2), indicating the favourable and comprehensive electrocatalytic activity of Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene for phenolic compound oxidation. Moreover, it's notable that the oxidation peaks presented distinct shifts to lower potential, revealing the electron transfer between Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene and phenolic compound.\\u003c/p\\u003e \\u003cp\\u003eTo further profile the unique electrocatalytic activity of Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene, several nanomaterials including MoS\\u003csub\\u003e2\\u003c/sub\\u003e nanosheets, WS\\u003csub\\u003e2\\u003c/sub\\u003e nanosheets, Fe\\u003csub\\u003e3\\u003c/sub\\u003eO\\u003csub\\u003e4\\u003c/sub\\u003e nanoparticles, TiO\\u003csub\\u003e2\\u003c/sub\\u003e nanoparticles and bulk Ti\\u003csub\\u003e3\\u003c/sub\\u003eAlC\\u003csub\\u003e2\\u003c/sub\\u003e were separately employed as constrast with 1-naphthol as substrate. It's notable that only Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene presented observable electrocatalytic activity for 1-naphthol oxidation, indicating that the unique electrocatalytic activity was the intrinsic property of Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene.\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003cp\\u003eTo quantitatively present the electrocatalytic activity of Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene for 1-naphthol oxidation, DPV curves were measured with the addition of 1-naphthol at different concentrations. As shown in Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig3\\\" class=\\\"InternalRef\\\"\\u003e3\\u003c/span\\u003eA, the peak current increased with the increasing concentration of 1-naphthol at low concentrations until reaching about 90 \\u0026micro;A. According to Faraday's laws of electrolysis and Michaelis-Menten equation, the fitting curve was achieved with hyperbola function, where the Michaelis constant was calculated to be 0.22 mM, indicating that Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene possessed strong affinity to 1-naphthol [\\u003cspan citationid=\\\"CR39\\\" class=\\\"CitationRef\\\"\\u003e39\\u003c/span\\u003e]. Moreover, the catalytic activity of Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene for 1-naphthol oxidation in homogeneous phase solution was explored using hydrogen peroxide as oxidant. Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig3\\\" class=\\\"InternalRef\\\"\\u003e3\\u003c/span\\u003eB displayed the real-time absorbance of the aqueous solutions at 387.5 nm, including 1-naphthol (curve a), mixture of 1-naphthol and hydrogen peroxide (curve b), mixture of 1-naphthol, hydrogen peroxide and Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene (curve c), respectively. It could be seen that the reaction ratio was significantly improved with the addition of Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene, indicating that Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene could efficiently catalyze the 1-naphthol oxidation by hydrogen peroxide as well [\\u003cspan citationid=\\\"CR40\\\" class=\\\"CitationRef\\\"\\u003e40\\u003c/span\\u003e] .\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003cp\\u003eTo survey the quantitative relation between electrocatalytic activity and amount of Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene, Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene solutions at different concentrations were employed to modify GCE to test the electrocatalytic activities for 1-naphthol oxidation, respectively. As shown in Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig3\\\" class=\\\"InternalRef\\\"\\u003e3\\u003c/span\\u003eC, the Faradic current increased linearly with the increasing concentration of Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene in low concentration range and reached a constant in high concentration range, indicating that the electrocatalytic activity was in direct proportion to the total modified area of Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene flakes but not further improved by the stacking of the flakes.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec11\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e3.3. Mechanism study for the catalytic activity of Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene\\u003c/h2\\u003e \\u003cp\\u003eThe first principle calculations are performed to explain the catalytic activity of Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene for 1-naphthol oxidation. The free energies of 1-naphthol adsorption were first calculated to determine whether 1-naphthol could be adsorbed by Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene. The optimized geometries of 1-naphthol on and Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene were given in Fig. S3 (see in Supplementary Material), which showed 1-naphthol molecule adsorbed on the plane surface of Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene in a \\\"lying-down\\\" or \\\"standing-up\\\" manner, with adsorption energies of -1.18476 and -0.75625 eV, respectively, indicating relative strong physical adsorption. As a result, the \\\"lying-down\\\" adsorption of 1-naphthol on Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene was critical in the electrocatalytic oxidation. To further investigate the origin of 1-naphthol adsorption, the differential charge density of 1-naphthol adsorbed on Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene was calculated. As shown in Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig3\\\" class=\\\"InternalRef\\\"\\u003e3\\u003c/span\\u003eD, the changes in charge density caused by 1-naphthol adsorption mainly came from hydroxyl group and oxygen atoms. Moreover, changes in charge density were also found on the aromatic rings, indicating that the aromatic structure played a role in leading to the \\\"lying-down\\\" adsorption mode of 1-naphthol on Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec12\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e3.4. Characterization of the DNA walking machine\\u003c/h2\\u003e \\u003cp\\u003eTo quantify the efficiency of DNA Walking machine, fluorescence dynamics experiments were carried out to verify it. Firstly, AuNPs and ssDNA-functionalized AuNPs were prepared according to previous literature with little adjustment [\\u003cspan citationid=\\\"CR41\\\" class=\\\"CitationRef\\\"\\u003e41\\u003c/span\\u003e]. In short, sodium citrate (3 mL, 1%) was added rapidly to the boiling solution of HAuCl\\u003csub\\u003e4\\u003c/sub\\u003e (100 mL, 1%). After the color changed from pale yellow to wine-red, the mixture was stopped heating and cooled to room temperature (RT) with continued stirring. The AuNPs had an average particle size of 13 nm and were stored at 4\\u0026deg;C for further use. The preparation method of ssDNA (support probe and walker-protect dsDNA) -functionalized AuNPs is as follows. Firstly, the 1428 \\u0026micro;L denature-supporting probe (2 \\u0026micro;M) and 72 \\u0026micro;L denature-walker-protect dsDNA (2 \\u0026micro;M) were mixed with 1 \\u0026micro;L acetic acid (500 mM, pH 5.2) and 0.5 \\u0026micro;L TCEP (100 mM) at RT for 1 h, respectively. Then the mixture was added into 1 mL AuNP solution, and the resultant solution was stored in a drawer at RT for at least 16 h. After 25 \\u0026micro;L Tris-acetate (500 mM, pH 8.2) was added to the mixture, 250 \\u0026micro;L NaCl (1 M) was dropwise added to the mixture every three hours (30, 40, 50, 60, 70 \\u0026micro;L were added respectively). Subsequently, the resulting mixture was stored in a drawer overnight. Lastly, the mixture was centrifuged (10 000 rpm, 10 min) to remove the excess reagents, and the red precipitate was washed and dispersed in DNA preparation solution for further use.\\u003c/p\\u003e \\u003cp\\u003eSecondly, fluorescence kinetics curve was shown in Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig4\\\" class=\\\"InternalRef\\\"\\u003e4\\u003c/span\\u003eA to prove the cutting efficiency of Nt.BsmAI nicking endonuclease (Nt.BsmAI). The curve a, b and c showed corresponding changes when different concentrations of target BCR/ABL fusion gene and 10 U Nt.BsmAI were added into the ssDNA (support probe and walker-protect dsDNA) -functionalized AuNPs solution, respectively. Curve d was the blank control. The slope of the curve reflects the reaction rate of enzyme shearing, which is correlated with the concentration of target gene. As can be seen from the figure, the reaction rate is fast. At the same time, when the reaction reached 2 hours, the shearing enzyme still did not reach the maximum shearing value, which means that the shearing enzyme has not been completely reacted. Thus, the cutting efficiency of the Nt.BsmAI is excellent. In addition, the time of releasing hairpin structure DNA can also be known from the curve. The sharply rising stage in the curve mainly the enzymatic cleaving on the prehybridized Support DNA-Walker. It can be seen from the figure that it takes about 6-8 minutes to cleave and release the hairpin. The slope of tangent at t = 0s was calculated with the fitting curves indicating the cleaving rate was highly related to the concentration of substrate in Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig4\\\" class=\\\"InternalRef\\\"\\u003e4\\u003c/span\\u003eB. The slope of tangent at t = 0s and logarithmic value of target BCR/ABL fusion gene concentrations presents well linear dependence range from 2 pM to 2 \\u0026micro;Μ with pearson correlation coefficient of 0.99192, which is corresponding to the kinetic characteristic of first-order reaction.\\u003c/p\\u003e \\u003cp\\u003eThirdly, as for the time of hairpin structure intermediate DNAs hybridized with the thiolated capture DNA on the electrode surface to form the sensing interface, we also conducted relevant optimization experiments. As shown in Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig4\\\" class=\\\"InternalRef\\\"\\u003e4\\u003c/span\\u003eC, the time needed was only 1h.\\u003c/p\\u003e \\u003cp\\u003eLastly, we have consulted relevant literature, which shows that the amount of DNA loading on 15 nm gold nanoparticles is 20-30 when the concentration of NaCl is 140mM [\\u003cspan citationid=\\\"CR42\\\" class=\\\"CitationRef\\\"\\u003e42\\u003c/span\\u003e]. According to the concentration and dosage of AuNPs, the amount of ssDNA fabricated on the interface of this sensor was calculated to be about 1\\u0026times;10\\u003csup\\u003e10\\u003c/sup\\u003e.\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec13\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e3.5. Optimization of experimental conditions\\u003c/h2\\u003e \\u003cp\\u003eTo achieved optimal analytical performance of the biosensor, some critical experimental conditions were optimized, including the ratio of walker probe to support probe, the cleaving time of Nt.BsmAI nicking endonuclease, the pH of DEA buffer and the cleaving temperature of Nt.BsmAI nicking endonuclease. As shown in Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig5\\\" class=\\\"InternalRef\\\"\\u003e5\\u003c/span\\u003e, optimal ratio of walker probe to support probe, cleaving time, pH of DEA buffer and temperature were achieved to be 1:20, 120 min, 9.6 and 37\\u0026deg;C, respectively.\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec14\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e3.6. Analytical performance of the proposed electrochemical biosensor\\u003c/h2\\u003e \\u003cp\\u003eTo estimate analytical performance of the biosensor, the current responses toward BCR/ABL fusion gene at different concentrations were recorded under the optimal conditions through DPV measurements. As shown in Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig6\\\" class=\\\"InternalRef\\\"\\u003e6\\u003c/span\\u003eA, the detection signal increased with the increasing concentration of target BCR/ABL fusion gene. The corresponding calibration plots of the peak currents showed a strong linear relationship to the logarithm value of target BCR/ABL fusion gene concentrations range from 0.2 fM to 20 nΜ with pearson correlation coefficient of 0.99836 (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig6\\\" class=\\\"InternalRef\\\"\\u003e6\\u003c/span\\u003eB). The linear regression equation was \\u003cem\\u003eI\\u003c/em\\u003e = 1.00012 \\u0026times; lg (\\u003cem\\u003ec/\\u003c/em\\u003epM) + 11.23074 (\\u003cem\\u003ec\\u003c/em\\u003e and \\u003cem\\u003eI\\u003c/em\\u003e stood for the concentration of target BCR/ABL fusion gene and corresponding peak current value, respectively). The limit of detection was obtained based on three times the average standard deviation corresponding to blank sample detection, which was calculated to be 0.05 fM. Comparisons of this biosensor with some reported works for BCR/ABL fusion gene detection are shown in Table S2, which highlighted the excellent sensitivity of this method in BCR/ABL fusion gene detection due to the cascading catalytic strategy and DNA walking machine for signal amplification.\\u003c/p\\u003e \\u003cp\\u003eMoreover, the specificity of the biosensor was evaluated by using 3 different DNA oligonucleotides as references, including a single-base-mismatched strand (B1), a two-base-mismatched strand (B2) and a noncomplementary strand (B3), all at concentrations of 20 fM. As depicted in Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig6\\\" class=\\\"InternalRef\\\"\\u003e6\\u003c/span\\u003eC, the response signals of the single-base-mismatched strand and two-base-mismatched strand were much lower than the response signal of the target, revealing the good capacity of the biosensor to distinguish base-mismatch. The response signal of noncomplementary sequences were approximate to the blank solution, indicating that the biosensor presented good selectivity for DNA detection.\\u003c/p\\u003e \\u003cp\\u003eTo evaluate the stability of the proposed biosensor, the modified electrodes were stored at 4\\u0026deg;C before use. As presented in Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig6\\\" class=\\\"InternalRef\\\"\\u003e6\\u003c/span\\u003eD, there were no obvious differences during the first 5 days of storage, and the current changes were less than 1.58%. After 20 days of storage, the designed biosensor retained 89.40% of its initial current response, indicating that the proposed biosensor offers satisfactory stability for target BCR/ABL detection.\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec15\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e3.7. Detection of BCR/ABL fusion gene in human serum samples\\u003c/h2\\u003e \\u003cp\\u003eTo further validate the applicability of the biosensor to complex biological matrix in clinical application, different concentrations of target BCR/ABL fusion gene were added to 10-fold-diluted clinical serum samples and tested with the proposed biosensor. The detection results of BCR/ABL fusion gene in human serum samples are summarized in Table S3. Satisfactory recovery values were obtained ranging from 93.60\\u0026ndash;110.42% with relative standard deviations (RSD) between 0.27% and 0.64%. In addition, we extracted RNA from clinical serum of BCR/ABL positive patients using spin columns CB3 according to the manufacture's protocol and tested with the proposed biosensor. The detection concentrations of BCR/ABL were compared with clinical results (by reverse transcription PCR), which were summarized in Table S4. It could be seen that the proposed biosensor achieved results well matched with the clinical assay, manifesting the application potential of the proposed biosensor in clinical diagnosis.\\u003c/p\\u003e \\u003c/div\\u003e\"},{\"header\":\"4. Conclusions\",\"content\":\"\\u003cp\\u003eIn summary, this work reported the unique catalytic activity of Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene nanozyme for phenols oxidation and the application to electrochemical biosensor for BCR/ABL fusion gene detection. The catalytic activity originated from the adsorption capacity of the Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene plane surface towards 1-naphthol. Theoretic calculation revealed the mechanism that relatively strong interaction was existed between the surface oxygen atoms of Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene and the phenolic hydroxyl groups. Moreover, the catalytic activity was strictly proportional to the cover area of the MXene fakes but independent to the number of stacking layers, which was quite different from traditional nanozymes and significant benefited the quality control of biosensor. Ultrasensitive detection was achieved with the proposed biosensor, which provided a promising analytical tool for the clinical diagnosis of CML.\\u003c/p\\u003e\"},{\"header\":\"Declarations\",\"content\":\"\\u003cp\\u003e\\u003cstrong\\u003eAcknowledgments\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThis research was supported by the National Natural Science Foundation of China (Grant Nos. 31071093, 31170129, and 31200064), China Postdoctoral Science Foundation (2020M683259) and the Science and Technology Planning Project of Yuzhong District of Chongqing city, China (No. 20140119).\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eAuthors\\u0026rsquo; contributions\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eRongjun Yu: Conceptualization, Methodology, Software, Investigation, Data Curation, Writing-Original Draft.\\u0026nbsp;Jian Xue: Software, Investigation, Project Administration.\\u0026nbsp;Yang Wang: Formal Analysis, Software.\\u0026nbsp;Jingfu Qiu: Supervision, Methodology, Data Curation, Administration. Xinyi Huang: Methodology, Software, Investigation, Project Administration. Anyi Chen: Writing-Review \\u0026amp; Editing. Jianjiang Xue: Funding Acquisition, Supervision, Administration, Data Curation.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eAvailability of data and materials\\u0026nbsp;\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eAll data generated or analyzed during this study are included in this article and the Additional Information.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eConsent for publication\\u0026nbsp;\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eAll authors of this study agreed to publish.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eCompeting interests\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe authors declare no competing financial interests.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eAuthor details\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cem\\u003e\\u003csup\\u003e1\\u0026nbsp;\\u003c/sup\\u003e\\u003c/em\\u003e\\u003cem\\u003eDepartment of Clinical Laboratory, University-Town Hospital of Chongqing Medical University, Chongqing, 401331, China.\\u003csup\\u003e2\\u003c/sup\\u003e\\u003c/em\\u003e \\u003cem\\u003eSchool of Public Health and Management, Chongqing Medical University, Chongqing, 400016, China.\\u003csup\\u003e3\\u003c/sup\\u003e\\u003c/em\\u003e\\u003cem\\u003e\\u0026nbsp;Department of Clinical Laboratory, First Affiliated Hospital of Guangxi University of Chinese Medicine, Nanning, 530023, China.\\u003c/em\\u003e\\u003c/p\\u003e\"},{\"header\":\"References\",\"content\":\"\\u003col\\u003e\\u003cli\\u003e\\u003cspan\\u003eKimmel DW, LeBlanc G, Meschievitz M, E,Cliffel DE. 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Maximizing DNA Loading on a Range of Gold Nanoparticle Sizes. Anal Chem. 2006;78:8313\\u0026ndash;8.\\u003c/span\\u003e\\u003c/li\\u003e\\u003c/ol\\u003e\"},{\"header\":\"scheme\",\"content\":\"\\u003cp\\u003eScheme 1 is available in the Supplemental Files section\\u003c/p\\u003e\"}],\"fulltextSource\":\"\",\"fullText\":\"\",\"funders\":[],\"hasAdminPriorityOnWorkflow\":false,\"hasManuscriptDocX\":true,\"hasOptedInToPreprint\":true,\"hasPassedJournalQc\":\"\",\"hasAnyPriority\":false,\"hideJournal\":false,\"highlight\":\"\",\"institution\":\"\",\"isAcceptedByJournal\":true,\"isAuthorSuppliedPdf\":false,\"isDeskRejected\":\"\",\"isHiddenFromSearch\":false,\"isInQc\":false,\"isInWorkflow\":false,\"isPdf\":false,\"isPdfUpToDate\":true,\"isWithdrawnOrRetracted\":false,\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"journal-of-nanobiotechnology\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":false,\"externalIdentity\":\"jnan\",\"sideBox\":\"Learn more about [Journal of Nanobiotechnology](http://jnanobiotechnology.biomedcentral.com)\",\"snPcode\":\"12951\",\"submissionUrl\":\"https://submission.nature.com/new-submission/12951/3\",\"title\":\"Journal of Nanobiotechnology\",\"twitterHandle\":\"@BioMedCentral\",\"acdcEnabled\":true,\"dfaEnabled\":true,\"editorialSystem\":\"em\",\"reportingPortfolio\":\"BMC/SO AJ\",\"inReviewEnabled\":true,\"inReviewRevisionsEnabled\":true},\"keywords\":\"Ti3C2Tx MXene, Nanozyme, Electrocatalysis, Cascading catalytic amplification, Electrochemical biosensor\",\"lastPublishedDoi\":\"10.21203/rs.3.rs-965061/v2\",\"lastPublishedDoiUrl\":\"https://doi.org/10.21203/rs.3.rs-965061/v2\",\"license\":{\"name\":\"CC BY 4.0\",\"url\":\"https://creativecommons.org/licenses/by/4.0/\"},\"manuscriptAbstract\":\"\\u003cp\\u003eIn this work, Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene was identified as efficient nanozyme with area-dependent electrocatalytic activity in oxidation of phenolic compounds, which originated from the strong adsorption effect between the phenolic hydroxyl group and the oxygen atom on the surface of Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene flake. On the basis of the novel electrocatalytic activity, Ti\\u003csub\\u003e3\\u003c/sub\\u003eC\\u003csub\\u003e2\\u003c/sub\\u003eT\\u003csub\\u003ex\\u003c/sub\\u003e MXene was combined with alkaline phosphatase to construct a novel cascading catalytic amplification strategy using 1-naphthyl phosphate (1-NPP) as substrate, thereby realizing efficient electrochemical signal amplification. Taking advantage of the novel cascading catalytic amplification strategy, an electrochemical biosensor was fabricated for BCR/ABL fusion gene detection, which achieved excellent sensitivity with linear range from 0.2 fM to 20 nM and limit of detection down to 0.05 fM. This biosensor provided a promising tool for ultrasensitive fusion gene detection in early diagnosis of chronic myelogenous leukemia and acute lymphocytic leukemia. Moreover, the manageable catalytic activity of MXene broke a path for developing nanozymes, which possessed enormous application potential in not only electrochemical analysis but also the extensive fields including organic synthesis, pollutant disposal and so on.\\u003c/p\\u003e\",\"manuscriptTitle\":\"Novel Ti3C2Tx MXene Nanozyme with Manageable Catalytic Activity and Application to Electrochemical Biosensor\",\"msid\":\"\",\"msnumber\":\"\",\"nonDraftVersions\":[{\"code\":2,\"date\":\"2022-02-15 23:00:36\",\"doi\":\"10.21203/rs.3.rs-965061/v2\",\"editorialEvents\":[{\"type\":\"communityComments\",\"content\":0},{\"type\":\"decision\",\"content\":\"Accept\",\"date\":\"2022-02-20T22:30:39+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"editorInvitedReview\",\"content\":\"\",\"date\":\"2022-01-31T13:19:50+00:00\",\"index\":0,\"fulltext\":\"\"},{\"type\":\"reviewersInvited\",\"content\":\"\",\"date\":\"2022-01-30T21:18:01+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"editorInvited\",\"content\":\"Journal of Nanobiotechnology\",\"date\":\"2022-01-20T21:47:28+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"editorAssigned\",\"content\":\"\",\"date\":\"2022-01-20T08:56:29+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"submitted\",\"content\":\"Journal of Nanobiotechnology\",\"date\":\"2022-01-16T08:49:42+00:00\",\"index\":\"\",\"fulltext\":\"\"}],\"status\":\"published\",\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"journal-of-nanobiotechnology\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":false,\"externalIdentity\":\"jnan\",\"sideBox\":\"Learn more about [Journal of Nanobiotechnology](http://jnanobiotechnology.biomedcentral.com)\",\"snPcode\":\"12951\",\"submissionUrl\":\"https://submission.nature.com/new-submission/12951/3\",\"title\":\"Journal of Nanobiotechnology\",\"twitterHandle\":\"@BioMedCentral\",\"acdcEnabled\":true,\"dfaEnabled\":true,\"editorialSystem\":\"em\",\"reportingPortfolio\":\"BMC/SO AJ\",\"inReviewEnabled\":true,\"inReviewRevisionsEnabled\":true}}],\"origin\":\"\",\"ownerIdentity\":\"2d574d5c-0a11-4595-85b0-d095224cd586\",\"owner\":[],\"postedDate\":\"February 15th, 2022\",\"published\":true,\"recentEditorialEvents\":[],\"rejectedJournal\":[],\"revision\":\"\",\"amendment\":\"\",\"status\":\"published-in-journal\",\"subjectAreas\":[],\"tags\":[],\"updatedAt\":\"2022-03-09T12:00:34+00:00\",\"versionOfRecord\":{\"articleIdentity\":\"rs-965061\",\"link\":\"https://doi.org/10.1186/s12951-022-01317-9\",\"journal\":{\"identity\":\"journal-of-nanobiotechnology\",\"isVorOnly\":false,\"title\":\"Journal of Nanobiotechnology\"},\"publishedOn\":\"2022-03-09 12:00:34\",\"publishedOnDateReadable\":\"March 9th, 2022\"},\"versionCreatedAt\":\"2022-02-15 23:00:36\",\"video\":\"\",\"vorDoi\":\"10.1186/s12951-022-01317-9\",\"vorDoiUrl\":\"https://doi.org/10.1186/s12951-022-01317-9\",\"workflowStages\":[]},\"version\":\"v2\",\"identity\":\"rs-965061\",\"journalConfig\":\"researchsquare\"},\"__N_SSP\":true},\"page\":\"/article/[identity]/[[...version]]\",\"query\":{\"redirect\":\"/article/rs-965061\",\"identity\":\"rs-965061\",\"version\":[\"v2\"]},\"buildId\":\"7rjqhiLT3MXkJMwkYKINL\",\"isFallback\":false,\"isExperimentalCompile\":false,\"dynamicIds\":[84888],\"gssp\":true,\"scriptLoader\":[]}","source_license":"CC-BY-4.0","license_restricted":false}