Ultrasound-pretreatment combined with Ti3C2-TiO2-AuNPs enhancing the electrogenerated chemiluminescence of the air-saturated luminol for exosomes detection

Huixin Zhang; Xin Zhou; Feifei Zhang; Jianfei Xia; Zonghua Wang

doi:10.1016/j.ultsonch.2023.106330

. 2023 Feb 13;94:106330. doi: 10.1016/j.ultsonch.2023.106330

Ultrasound-pretreatment combined with Ti₃C₂-TiO₂-AuNPs enhancing the electrogenerated chemiluminescence of the air-saturated luminol for exosomes detection

Huixin Zhang ¹, Xin Zhou ¹, Feifei Zhang ¹, Jianfei Xia ¹, Zonghua Wang ^1,^⁎

PMCID: PMC9969320 PMID: 36805412

Graphical abstract

The cavitation effect of high-intensity focused ultrasound could generate the H₂O₂ and O₂^–• in situ. Ti₃C₂-TiO₂-AuNPs absorbed and catalyzed H₂O₂ to produce more O₂^–•. The synergistic effect leads to a great enhancement of the ECL emission for the air-saturated luminol.

Keywords: Ultrasound-pretreatment, Ti₃C₂-TiO₂-AuNPs, Electrogenerated chemiluminescence, The air-saturated luminol, ROS

Highlights

•
High-intensity focused ultrasound triggered the ECL of the air-saturated luminol.
•
A novel probe of Ti₃C₂-TiO₂-AuNPs was synthetized.
•
The Ti₃C₂-TiO₂-AuNPs could absorb and catalyze the H₂O₂ to O₂^–•.
•
Through combination of ultrasound and probe, the exosomes were detected sensitively.

Abstract

It is still a great challenge to develop effective strategies to improve the low electrogenerated chemiluminescence (ECL) of air-saturated luminol. Herein, the synergistic effects of Ti₃C₂-TiO₂-AuNPs nano hybrid and high-intensity focused ultrasound pretreatment (ultrasound-pretreatment) were used to significantly improve the ECL emission of the air-saturated luminol, and the mechanism was proposed. The ultrasound-pretreatment as a green method with the cavitation effect could form O₂^–• and H₂O₂ in situ as an initiator. TiO₂ and Au nanoparticles (AuNPs) were in situ decorated on the Ti₃C₂ surface to form Ti₃C₂-TiO₂-AuNPs, and it was proved as a highly efficient booster which could catalyze and aggregate H₂O₂ to the O₂^–•. The utilization rate of intermediates has been greatly improved. Exosomes as model targets can be sensitively detected by the ECL sensor. The detection limit was 195 particles μL⁻¹. The detection results of exosomes in actual samples are satisfactory. We believe that the ultrasound-pretreatment strategy could be extended to the sensitive detection in the biological sample.

1. Introduction

Luminol is a classic organic ECL emitter, which is favored because of its low oxidation potential, nontoxicity, and so on [1], [2], [3]. In the air-saturated luminol, dissolved oxygen with low toxicity and natural stability is a potential and green endogenous coreactant of luminol [4], [5], [6]. However, the low content and conversion rate of dissolved oxygen result unsatisfied ECL intensity of luminol. To solve the problem, coreactants are usually introduced to activate the air-saturated luminol to generate more the reactive oxygen species (ROS). So far, numerous species, such as, enzymes, nanomaterials (Cu₂O@AuNPs, Pt-Au@ZnO and Au-Ag-Pt heteronanostructures [7], [8], [9] and single-atom catalysts [10], [11], [12], [13]) have been developed as the accelerator to promote conversion of O₂ to ROS. The developing of green and effective means to boost the ECL strength of the air-saturated luminol is still a hot topic.

It should be noted that ultrasound, as a unique non-invasive means, has played a significant role in biochemical research and clinical treatment [14], [15]. Among them, high-intensity focused ultrasound has the inherent advantages of reducing off target damage and improving ultrasound dept, which is considered to be one of the most promising minimally invasive methods [16], [17]. High negative pressure in the ultrasound focus area can generate cavitation bubbles, then the cavitation bubbles oscillate and collapse. In the process of bubble shrinkage, the internal gas is compressed, resulting in the decomposition of water molecules and O₂ to generate ROS, such as O₂^–•, ¹O₂, H₂O₂ and so on [18], [19]. So far, only a few papers on the application of ultrasound in ECL have been reported [20], [21]. Our research group has done the related work in the field of ECL sensing, especially in improving selectivity and sensitivity by using nanomaterials [3], [5]. We speculated that the combination of ultrasound and nanomaterials will be a potential means to boost ECL signal of air-saturated luminol.

Titanium dioxide (TiO₂) has become an emerging material in recent years in the biosensing [22], [23], [24], [25]. It was reported that TiO₂ could catalyze the oxidation of H₂O₂ and aggregate ROS, such as O₂^–• and •OH owing to the band gap dependent catalytic performance of TiO₂, which contributes significantly to the ECL emission of luminol [26]. Recently, based on TiO₂ nano-hybrid, such as AuAg/TiO₂ [27], Att-TiO₂ [28], Fe₃O₄-TiO₂@NH₂ [29] with strong catalytic performance and good conductivity have attracted much attention in the ECL sensing. However, pre-synthesis of TiO₂ usually requires multi-step process and the synthesized TiO₂ has a large size, leading to small specific surface area and relatively few active sites of the nano-hybrid. Therefore, it is necessary to find an effective and easy synthesis method of TiO₂ with small size to exert its excellent performance in the ECL biosensing. In situ growth of TiO₂ on the nanomaterial could be a promising approach. Fortunately, Ti₃C₂ MXenes as a novel two-dimensional nanomaterial has rich exposed metal sites, excellent reducibility, catalysis and electrical conductivity [30], [31], [32], [33], [34], [35], [36], which can provide rich Ti source to generate the small size of TiO₂ in situ. Ti₃C₂ nanosheets can be used as support platform for TiO₂, which can effectively prevent the self-aggregation of TiO₂. We expected the Ti₃C₂-TiO₂ generated in situ has an effect of “two birds with one stone” to enhance the ECL response of the air-saturated luminol.

In the present work, we designed a ECL platform with synergistic effect to achieve the goal of sensitive determination of exosomes (Scheme 1). Ultrasound with ultrasonic cavitation effect was a pretreatment means and TiO₂ and AuNPs were generated in situ on Ti₃C₂ to form Ti₃C₂-TiO₂-AuNPs as nanoprobes. The Ti₃C₂-TiO₂-AuNPs nanoprobes can catalyze H₂O₂ and aggregate ROS generated from ultrasound-pretreatment via cavitation ingeniously to greatly boost the ECL emission of the air-saturated luminol. In the Scheme 1A, the Ti₃C₂-TiO₂-AuNPs nano hybrid was formed by the reducibility of Ti. The Scheme 1B shows the construction of ECL sensor for exosome detection. The CD63 aptamer2 (Apt2) was modified on sodium alginate (SA). Moreover, the Ti₃C₂-TiO₂-AuNPs nano hybrid conjugated with numerous CD63 aptamer1 (Apt1) via Au-S band was captured on the exosomes, and the assembled electrode was detected the exosomes in the luminol pretreated by ultrasound. Scheme 1C shows the illustration for the synergistic effect of ultrasound-pretreatment and Ti₃C₂-TiO₂-AuNPs for the air-saturated luminol. The ultrasound-pretreatment strategy could generate superoxide radical (O₂^–•) and H₂O₂ effectively. More importantly, Ti₃C₂-TiO₂-AuNPs nanoprobes not only catalyze H₂O₂ but also aggregate ROS generated in situ via ultrasound-pretreatment, which can short the diffusion distance and improve the intermediate utilization rate, leading the enhancement of the ECL emission for the air-saturated luminol. The designed ECL biosensor can detect exosomes even in serum, blood, and urine samples, thus proving that it is a promising method for the routine monitoring of exosomes in clinical diagnostics.

2. Experimental

2.1. Materials and reagents

Reagents, materials, instruments, preparation of Ti₃C₂, Ti₃C₂-TiO₂ and Ti₃C₂-TiO₂-AuNPs are shown in Supplementary Material.

2.2. Exosome extraction and characterization

The culture of MDA-MB-231 cells and the extraction of exosomes refer to our previous work [5]. The TEM image of exosome revealed that the size of approximately is 50 nm [37] (Fig. S1).

2.3. Construction of the ECL platform

The glassy carbon electrode (GCE) with a diameter of 3 mm was polished with 0.3 and 0.05 μm alumina powder by turns and ultrasonic cleaning with ethanol and deionized water respectively. 6 μL of SA solution was modified on the GCE to form SA/GCE. Then, SA/GCE was activated in NHS and EDC (200 mM, 800 mM, v/v = 1:1) at 37°C for 1 h. Then, the SA/GCE was incubated in CD63 Apt2 (1 μM) at 37°C for 2 h to obtain Apt2/SA/GCE. Afterwards, different concentrations of exosomes (100 μL) were incubated on the Apt2/SA/GCE at 37°C for 2 h to form exosomes/Apt2/SA/GCE. The exosomes/Apt2/SA/GCE was incubated in 35 μL Ti₃C₂-TiO₂-AuNPs-Apt1 nanoprobes for 2 h to obtain an ECL biosensor. Finally, the ECL measurement was performed in luminol solution with ultrasound-pretreatment to detect exosomes. In each of the above steps, the electrode needs to be cleaned with ultrapure water and dried with N₂ to remove non-specific adsorption.

2.4. Ultrasound -pretreatment and ECL measurement

In this experiment, ultrasound was used as a pretreatment means. The air-saturated luminol solution (50 μM) was pretreated with ultrasound (7.5 W) at 1 MHz for 5 min, and then ultrasound was stopped and removed. Next, the modified electrode (Ti₃C₂-TiO₂-AuNPs-Apt1/exosomes/Apt2/SA/GCE) was used to detect exosomes in the pretreatment luminol solution.

In this work, a three-electrode system was used, the working electrode was the modified electrode, the reference electrode was Ag/AgCl (saturated KCl solution), and the auxiliary electrode was a platinum wire. The scanning potential was set from 0 to 0.6 V, and the scanning speed was 0.1 V s⁻¹. The PMT was 700 V.

3. Results and discussion

3.1. Characterization of the samples

To prove the preparation of Ti₃C₂-TiO₂ and Ti₃C₂-TiO₂-AuNPs, transmission electron microscopy (TEM), high resolution transmission electron microscope (HRTEM), X-ray diffraction (XRD) and X-ray photoelectron spectroscopy (XPS) characterization and EDS analysis were conducted. In the TEM image, the average diameter of Ti₃C₂ nanosheets was approximately 500 nm with smooth surfaces (Fig. 1A). After oxidation, the layered structure of Ti₃C₂ was retained and TiO₂ particles formed on the Ti₃C₂ nanosheets in situ can be observed (Fig. 1B and Fig. S2). The Ti₃C₂-TiO₂ was also characterized by XRD (Fig. 1E). The strongest peak of (1 0 4) crystal plane disappeared, the peak of (0 0 2) moved slightly to the left because of expansion of the interlayer of Ti₃C₂, which indicated that Ti₃C₂ nanosheets have been successfully synthesized. Meanwhile, the specific diffraction peaks of (1 0 1), (0 0 4), (2 0 0) and (2 1 3) crystal planes of TiO₂ appeared and the (0 0 2) peak of Ti₃C₂ existed, indicating that the Ti₃C₂-TiO₂ was successfully synthesized.

Moreover, the XPS full spectrum of Ti₃C₂-TiO₂ was shown in Fig. S3A. In the C1s spectra of Ti₃C₂-TiO₂ (Fig. S3B), the peak value (284.7 eV) belonged to adventitious carbon (C-C). The C1s spectra at 281.75 and 285.9 eV correspond to C-Ti and C-O. The wide peak at 455–465 eV, which were related with low valence and high valence Ti species. The peaks at ∼455.4 eV and ∼461.5 eV belonged to Ti-C and Ti-O bonds (Fig. S3C). The formation of Ti₃C₂-TiO₂ nano hybrid could lead to the transition of Ti (II) to Ti (IV). Two typical peaks of O1s of Ti₃C₂-TiO₂ appeared, which were related with Ti-O and C-Ti-Ox (Fig. S3D). The results showed that TiO₂ could be in-situ grown on Ti₃C₂.

The TEM and SEM images of the Ti₃C₂-TiO₂-AuNPs were shown in Fig. 1C and Fig. S4A, indicating that the AuNPs were dispersed on the Ti₃C₂ nanosheets. Furthermore, Fig. 1D was the HRTEM image of Ti₃C₂-TiO₂-AuNPs, it clearly exhibited the lattice fringe spacings of 0.23 and 0.35 nm, belonged to the (1 0 3) plane of Ti₃C₂ and the (1 0 1) plane of anatase TiO₂. Meanwhile, a clear lattice fringe spacing of about 0.20 nm could be observed, which was consistent with the Au (1 1 1) plane. The characterization of Ti₃C₂-TiO₂-AuNPs by EDS analysis was also proved. Ti and O elements came from Ti₃C₂ and TiO₂, C element came from the Ti₃C₂. Au element came from the AuNPs (Fig. S4B). In addition, in the UV–vis spectra (Fig. S4C), comparing with curve a, the absorption peak of Ti₃C₂-TiO₂ displayed red shift slightly (curve b). In the curve c, due to the surface plasma absorption of AuNPs, a characteristic absorption peak appeared at 550 nm approximately. The formation of Ti₃C₂-TiO₂-AuNPs was also recorded by the digital photo (Fig. S4D). The Ti₃C₂-TiO₂ was well dispersed in water with dark green (a), the HAuCl₄ solution resulted in yellow (b). In contrast, the mixture of Ti₃C₂-TiO₂ and HAuCl₄ leaded to purplish red (c), indicating reduction of AuCl₄^- to form the Ti₃C₂-TiO₂-AuNPs. The EDS mapping of Ti₃C₂-TiO₂-AuNPs was shown in Fig. S5. C, O, Ti, F and Au elements appeared clearly, which could be more directly to demonstrate the successful synthesis of Ti₃C₂-TiO₂-AuNPs.

3.2. The effect of ultrasound-pretreatment and the Ti₃C₂-TiO₂-AuNPs

Firstly, the critical role of Ti₃C₂-TiO₂-AuNPs modified electrode in the air-saturated luminol with ultrasound-pretreatment for 5 min was studied. An ultralow ECL strength was gained on the bare GCE (Fig. 2A, curve a). After modification with Ti₃C₂, Ti₃C₂-TiO₂ and Ti₃C₂-AuNPs, Ti₃C₂-TiO₂-AuNPs, the ECL intensity was increased in turn (curve b, c, d and e). These results indicated that the Ti₃C₂-TiO₂-AuNPs nano hybrid showed the best electrocatalytic activity and conductivity than that of Ti₃C₂, Ti₃C₂-TiO₂ or Ti₃C₂-AuNPs. Amazingly, aiding by the ultrasound-pretreatment, the Ti₃C₂-TiO₂-AuNPs/GCE in the air-saturated luminol showed the highest ECL signal (curve f), which indicated that the ROS was generated in situ via the unique cavitation effect of ultrasound, which may be an important factor in enhancing ECL emission. In addition, the stability of the Ti₃C₂-TiO₂-AuNPs/GCE with ultrasound-pretreatment was studied under consecutive scanning for 10 cycles. These responses almost remained unchanged, and the RSD was 2.70% (Fig. 2B).

In addition to the ECL performance, the ECL enhancement mechanism was studied. As shown in the Fig. 2C, the Ti₃C₂-TiO₂-AuNPs/GCE exhibited a strong ECL emission in the air-saturated luminol with ultrasound-pretreatment, a low ECL strength was gained in the luminol which was saturated N₂ with ultrasound-pretreatment. These results showed that the dissolved oxygen was important in this system. It is well known that the O₂^–• was important as the intermediate, and it could oxidize luminol to 3-aminophthalate to generate ECL signal. Therefore, a commercially specific radical scavenger benzoquinone (BQ) for O₂^–• was introduced to confirm the nature of the involved reactive species. It was seen from the Fig. 2C, the addition of BQ induced 86 % quenching of the ECL signal, which confirmed that O₂^–• is mainly in the ROS generated by ultrasound-pretreatment and Ti₃C₂-TiO₂-AuNPs.

Since the reactive O₂^–• could yield H₂O₂ in the aqueous solution, we tended to confirm the existence of H₂O₂. The in-situ generation of H₂O₂ via ultrasound-pretreatment was validated by the H₂O₂ specific colorimetric method, in which HRP only catalyzed H₂O₂ to oxidize the substrate TMB to a colored product (TMB*, 650 nm). Compared with curve b in Fig. 2D, a specific absorbance was observed at about 650 nm by ultrasound-pretreatment (curve a), which confirmed the existence of H₂O₂. Moreover, the maximum absorption peak at 650 nm exhibited a linear fit with the H₂O₂ concentration (R² = 0.997) in the range of 10–50 μM (Fig. S6). According to the linear equation of A = -0.018 + 0.034C, the concentration of H₂O₂ generated by ultrasound-pretreatment was calculated as 26.47 μM.

The high-intensity focused ultrasound with the cavitation effect could form O₂^–• and H₂O₂ in situ as an initiator. Ti₃C₂-TiO₂-AuNPs was proved as a highly efficient booster, which could catalyze H₂O₂ to the O₂^–•. The synergistic effects of ultrasound-pretreatment and Ti₃C₂-TiO₂-AuNPs greatly improved the ECL intensity in the air-saturated luminol. The ECL mechanism for the application of ultrasound-pretreatment and Ti₃C₂-TiO₂-AuNPs in the air-saturated luminol was proposed in Scheme 1C and Eqs. (1) - (6). A large amount of luminol anions (LH^-) were electrochemically oxidized to luminol anionic radicals (L^•-) [Eq. (1)]. Dissolved O₂ could obtain e^- on the electrode to produce O₂^–• [Eq. (2)]. Ultrasound-pretreatment effectively generated H₂O₂ and O₂^–• via cavitation effect [Eq. (3)]. Ti₃C₂-TiO₂-AuNPs could absorb and catalyze the H₂O₂ to generate more O₂^–• [Eq. (4)]. Lastly, excited-state intermediate (AP*) was formed via the reaction of L^•- with O₂^–•, which returned to the ground state to obtain excellent ECL emission [Eq. (5), (6)].

L H^{-} - e^{-} \to L^{\cdot -} + H^{+}

(1)

O_{2} + e^{-} \to O_{2}^{- \cdot}

(2)

O_{2} \overset{Ultrasound}{\to} O_{2}^{- \cdot} + H_{2} O_{2}

(3)

H_{2} O_{2} \to_{catalyze a n d a b s o r b}^{T i_{3} C_{2} - T i O_{2} - A u N P s} m o r e O_{2}^{- \cdot} + H^{+}

(4)

L^{\cdot -} + O_{2}^{- \cdot} \to A P^{*} + N_{2}

(5)

A P^{*} \to A P + h v

(6)

3.3. Electrochemical characterizations of the ECL platform

Cyclic voltammetry (CV) was used to characterize the construction of this sensor. After modifying SA on bare electrodes, a smaller current (Fig. 3A, b) than that of the bare GCE (Fig. 3A, a) was shown distinctly. After Apt2 (c) and exosomes (curve d) were incubated, the peak current gradually decreased, because the non-conductive material retarded the electron transfer between the GCE and electroactive material. Finally, when the nanoprobes were connected to the surface of exosomes, the current significantly decreased, owing to the electron transport on electrode surface was blocked by the negatively charged nanoprobes (curve e). The assembly process of this biosensor was also characterized via electrochemical impedance spectroscopy (Fig. 3B). The SA, Apt2, exosomes and Ti₃C₂-TiO₂-AuNPs-Apt1 were modified on the GCE in turn, the arch became bigger gradually, indicating that charge-transfer resistance increased gradually. The above results indicated that the electrode assembly was successful.

3.4. Detection of exosomes

Exosomes are vesicles derived from cells (30–200 nm), which carry rich nucleic acids, proteins and lipids for intercellular communication. Because exosomes are highly related to the changes in physiological and pathological conditions for the diseases, especially cancer [38], they are widely considered as potential biomarkers for biological analysis and disease treatment.

The ECL intensity towards different concentrations of exosomes was studied under the optimal experimental conditions (Fig. S7). The ECL response increased with the concentration of exosomes increased from 5.0 × 10² to 1.0 × 10⁵ particles μL⁻¹ with a good linear relationship (Fig. 4A). The linear equation is I = -10411.56 + 4839.51LgC. The limit of detection (LOD) was 195 particles μL⁻¹, which was calculated via formula of LOD=(3σ-b)/k (σ is the background standard deviation, b is the intercept of the linear regression equation, and k is the slope of the linear regression equation), and the squared correlation coefficient (R²) was 0.996. These results showed that this ECL platform had excellent performance for the exosome detection.

Fig. 4 — (A) ECL response of this platform with different concentrations of exosomes (a-f: 5.0 × 10², 1.0 × 10³, 5.0 × 10³, 1.0 × 10⁴, 5 × 10⁴, 1.0 × 10⁵ particles μL⁻¹, respectively) in the air-saturated luminol with ultrasound-pretreatment. (B) The corresponding linear calibration curve.

3.5. Stability and reproducibility of this platform

Moreover, measuring the ECL responses for 10 cycles was used to prove the stability, the ECL signals did not change significantly (Fig. 5). The reproducibility was evaluated via five Ti₃C₂-TiO₂-AuNPs-Apt1/exosomes/Apt2/SA/GCE, which was prepared by the same modification method. The RSD was 1.50% with 1.0 × 10⁴ particles μL⁻¹ exosomes for the ECL response of five electrodes, showing that this platform had excellent reproducibility.

3.6. Sample testing

The feasibility of the constructed sensors to detect target in actual samples is important of clinical diagnosis. Exosomes (1.0 × 10³ and 5.0 × 10³ particles μL⁻¹) were added into serum, urine and blood samples for recovery experiments, respectively. The recovery rates are between 96.22% and 108.33% with the RSD of 1.67% to 5.51% (Table S1), showing that this biosensor had excellent anti-interference ability in complex matrix and showed potential in clinical sample detection.

4. Conclusion

A novel ECL sensor for detecting exosomes was designed through combining ultrasound-pretreatment with Ti₃C₂-TiO₂-AuNPs nano hybrid. Ultrasound-pretreatment as green and effective means could generate H₂O₂ and O₂^–• in situ via ultrasonic cavitation effect. Moreover, Ti₃C₂-TiO₂-AuNPs nano hybrid with strong catalytic performance could catalyze and aggregate ROS to generate more O₂^–•, thereby enhancing ECL emission of the air-saturated luminol. As a proof of concept, the designed ECL platform was successfully used for exosome detection without additional co-reaction reagent. The LOD was 195 particles μL⁻¹. More importantly, this sensor could be used in detecting exosomes in serum, urine and blood. This green and concise method provides possibilities for the application of the air-saturated luminol.

5. Author statement

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Neither the entire article nor any part of its content has been published or has been accepted elsewhere and is not submitted to any other journals.

Declaration of Competing Interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Acknowledgments

This work was financially supported by the Natural Science Foundation of Shandong Province (ZR2020MB063) and the Taishan Scholar Program of Shandong Province (ts201511027), China.

Footnotes

^{Appendix A}

Supplementary data to this article can be found online at https://doi.org/10.1016/j.ultsonch.2023.106330.

Appendix A. Supplementary data

The following are the Supplementary data to this article:

Supplementary data 1

mmc1.docx^{(2.5MB, docx)}

Data availability

No data was used for the research described in the article.

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Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Supplementary Materials

Supplementary data 1

mmc1.docx^{(2.5MB, docx)}

Data Availability Statement

No data was used for the research described in the article.

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[b0130] 26.Ma X.G., Wang C., Wu F.X., Guan Y.R., Xu G.B. TiO2 Nanomaterials in Photoelectrochemical and Electrochemiluminescent Biosensing. Topics Curr. Chem. 2020;37:28. doi: 10.1007/s41061-020-0291-y. [DOI] [PubMed] [Google Scholar]

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[b0140] 28.Wang Y.Z., Zhong H., Li X.R., Liu G.Q., Yang K., Ma M., Zhang L.L., Yin J.Z., Cheng Z.P., Wang J.K. Nonenzymatic electrochemiluminescence glucose sensor based on quenching effect on luminol using attapulgite-TiO2. Sensor. Actuat. B-Chem. 2016;230:449–455. [Google Scholar]

[b0145] 29.Wu X.P., Zhong X., Chai Y.Q., Yuan R. Electrochemiluminescence acetylcholine biosensor based on biofunctional AMs-AChE-ChO biocomposite and electrodeposited graphene-Au-chitosan nanocomposite. Electrochim. Acta. 2014;147:735–742. [Google Scholar]

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[b0165] 33.Zhang H.X., Wang Z.H., Wang F., Zhang Y.M., Wang H.Y., Liu Y. Ti3C2 MXene mediated prussian blue in situ hybridization and electrochemical signal amplification for the detection of exosomes. Talanta. 2021;224 doi: 10.1016/j.talanta.2020.121879. [DOI] [PubMed] [Google Scholar]

[b0170] 34.Tang R., Xiong S., Gong D., Deng Y., Wang Y., Su L., Ding C., Yang L., Liao C. Ti3C2 2D MXene: Recent progress and perspectives in photocatalysis. ACS Appl. Mater. Interfaces. 2020;12(51):56663–56680. doi: 10.1021/acsami.0c12905. [DOI] [PubMed] [Google Scholar]

[b0175] 35.Wang L., Zhang H.X., Zhuang T.T., Liu J.X., Sojic N., Wang Z.H. Sensitive electrochemiluminescence biosensing of polynucleotide kinase using the versatility of two-dimensional Ti3C2TX MXene nanomaterials. Anal. Chim. Acta. 2022;1191 doi: 10.1016/j.aca.2021.339346. [DOI] [PubMed] [Google Scholar]

[b0180] 36.Wei Z., Zhang H., Wang Z. High-intensity focused ultrasound combined with Ti3C2-TiO2 to enhance electrochemiluminescence of luminol for the sensitive detection of polynucleotide kinase. ACS Appl. Mater. Interfaces. 2023;15(3):3804–3811. doi: 10.1021/acsami.2c19539. [DOI] [PubMed] [Google Scholar]

[b0185] 37.Cynthia A., Sepideh M., Houman K., Milad A., Mohammad S.S.Z., Mahsa J., Dariush S., Behzad B., Masood D., Tohid K. Docosahexaenoic acid (DHA) inhibits pro-angiogenic effects of breast cancer cells via down-regulating cellular and exosomal expression of angiogenic genes and microRNAs. Life Sci. 2020;258 doi: 10.1016/j.lfs.2020.118094. [DOI] [PubMed] [Google Scholar]

[b0190] 38.Wan S., Zhang L., Wang S., Liu Y., Wu C., Cui C., Sun H., Shi M., Jiang Y., Li L., Qiu L., Tan W. Molecular recognition-based DNA nanoassemblies on the surfaces of nanosized exosomes. J. Am. Chem. Soc. 2017;139(15):5289–5292. doi: 10.1021/jacs.7b00319. [DOI] [PMC free article] [PubMed] [Google Scholar]

PERMALINK

Ultrasound-pretreatment combined with Ti3C2-TiO2-AuNPs enhancing the electrogenerated chemiluminescence of the air-saturated luminol for exosomes detection

Huixin Zhang

Xin Zhou

Feifei Zhang

Jianfei Xia

Zonghua Wang