CN114088793A - Organic electrochemical transistor sensor and preparation method and application thereof - Google Patents
Organic electrochemical transistor sensor and preparation method and application thereof Download PDFInfo
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Abstract
The invention relates to a preparation method of an organic electrochemical transistor sensor, which comprises the following steps: preparing two-dimensional layered transition metal carbide; preparing a nano composite material; preparing an organic electrochemical transistor sensor; modifying the organic electrochemical transistor sensor. The invention also relates to the application of the organic electrochemical transistor sensor in high-sensitivity detection of survivin. The organic electrochemical transistor sensor can carry out high-sensitivity detection on the survivin.
Description
Technical Field
The invention relates to the technical field of chemical detection, in particular to an organic electrochemical transistor sensor and a preparation method and application thereof.
Background
Survivin (survivin), a 16.5-kD protein, belongs to the family of apoptosis Inhibitors (IAPs) and is an important oncogenic protein highly expressed in osteosarcomas. Normal cells do not require survivin, however, survivin is of great significance for cell division and apoptosis inhibition of osteosarcoma tumor cells. In addition to the functions of regulating cell division and inhibiting apoptosis, the increase of survivin protein level has been considered as a new potential osteosarcoma diagnostic biomarker, which can be used as a reference index for determining the pathological classification of osteosarcoma.
Various methods have been reported for detecting survivin, such as an enzyme-linked immunosorbent assay and a plasmon resonance assay, and then the methods have disadvantages of expensive instruments, complicated operation, and limited sensitivity and specificity, and thus it is necessary to develop a convenient method with high sensitivity and high specificity for detecting survivin.
Organic electrochemical transistors (OECTs) have shown great potential and advantage in biological and biochemical sensing applications due to their unique advantages, such as high sensitivity, low operating voltage, biocompatibility and flexibility, and simplified manufacturing processes, enabling their operating environment in water. OECT biosensors based on poly (3, 4-ethylenedioxythiophene) poly (styrenesulfonate) (PEDOT: PSS) have been widely used to detect bacteria, dopamine, DNA, cells and ions, indicating that OECT sensitivity is superior to those of conventional methods.
Ti3C2Tx is a two-dimensional layered early transition metal carbide/carbonitride (MXene) [ Tx denotes surface termination function (e.g. F, O and OH)]Showing great promise, MXene is widely used in biosensor design and manufacture, including the following advantages: 1) up to 9880S cm-1High excellent metal conductivity; 2) up to 1500F cm-3High capacitance, intercalation/deintercalation and redox reactions; 3) ti3C2The hydrophilic surface of Tx MXene makes it suitable for various solution processes such as spray coating and spin coating for manufacturing biosensor devices in aqueous environments. Compared with nano materials such as PPy, PANI, rGO, C-dots, AuNPs, ZnO and the like, the unique embedding/de-embedding nano structure enables MXene @ PEDOT: PSS as an ideal binder candidate synergistically enables high performance OECT biosensors with high volumetric capacitance. Therefore, the survivin can be effectively detected by constructing an organic electrochemical transistor.
Disclosure of Invention
The invention aims to provide an organic electrochemical transistor sensor and a preparation method and application thereof, aiming at the defects in the prior art.
In order to achieve the purpose, the invention adopts the technical scheme that:
the first aspect of the invention provides a method for preparing an organic electrochemical transistor sensor, which comprises the following steps:
s1, preparing two-dimensional layered transition metal carbide: selective etching of Ti in aqueous LiF/HCl3AlC2The aluminum layer in (1) to obtain Ti3C2Tx MXene；
S2, preparing the nano composite material: ti prepared in step S13C2Mixing Tx MXene with PEDOT (PSS) in a ratio of 1:2-2:1 to obtain MXene/PEDOT (PSS);
s3, preparing an organic electrochemical transistor sensor: washing a natural oxide p + silicon substrate with acetone, isopropanol and water in sequence, drying, and patterning through a photoresist and a shadow mask to obtain the organic electrochemical transistor sensor;
s4, modifying the organic electrochemical transistor sensor: the organic electrochemical transistor sensor prepared in the step S3 is coated with MXene/PEDOT: PSS prepared in the step S2 on its surface, and a survivin antibody is modified on the gate electrode.
Preferably, step S1 includes:
LiF was dissolved in HCl under magnetic stirring, and Ti was added slowly3AlC2Stirring at 20-60 deg.C for a period of time, repeatedly washing with deionized water for 3-9 times until pH reaches 6-7, and drying to obtain Ti3C2Tx MXene。
Preferably, the patterning in step S3 includes:
the source electrode and the drain electrode of Cr/Au are deposited on p by thermal evaporation through a channel structure+Cleaning the silicon substrate by deionized water, drying by a nitrogen gun, and baking at the temperature of 100-140 ℃ for 15-45 seconds.
Preferably, the working pressure of Cr thermal evaporation deposition is 1.2Pa, and the thickness is 10 nm.
Preferably, the working pressure of Au thermal evaporation deposition is 0.5Pa, and the thickness is 50 nm.
Preferably, the parameters for the modification of the survivin antibody in step S4 include: the pH value is 7.0; the time is 30 minutes; the temperature is 37 ℃; the concentration of survivin antibody protein was 10. mu.g/mL.
A second aspect of the present invention is to provide an organic electrochemical transistor sensor fabricated using the fabrication method as described above.
A third aspect of the present invention provides the use of an organic electrochemical transistor sensor as described above in the highly sensitive detection of survivin.
By adopting the technical scheme, compared with the prior art, the invention has the following technical effects:
the organic electrochemical transistor sensor can carry out high-sensitivity detection on the survivin.
Drawings
FIG. 1a is a schematic diagram of the preparation process of MXene/PEDOT: PSS nanocomposite;
FIG. 1b is a schematic diagram of the structure of an organic electrochemical transistor sensor;
FIGS. 1c and 1d are circuit diagrams characterizing an organic electrochemical transistor sensor in a liquid electrolyte by equivalent electric double layer capacitance;
FIG. 1e is a graph of XRD analysis of MXene and MXene/PEDOT: PSS nanocomposites;
FIG. 1f is a SEM top view of MXene/PEDOT: PSS nanocomposite;
FIG. 1g is an SEM side view of MXene/PEDOT: PSS nanocomposite;
where CGE is the capacitance of the gate/electrolyte interface and CEC is the capacitance of the electrolyte/channel interface;
FIG. 2a is a schematic flow diagram of layer-by-layer assembly on a gate electrode in an organic electrochemical transistor sensor;
FIG. 2b is a graph of EIS results for an organic electrochemical transistor sensor;
FIG. 2c is a graph of cyclic voltammetry results for an organic electrochemical transistor sensor;
FIG. 2d is a graph showing the results of the contact angles of naked gold, Au/MAA/anti-survivin protein/BSA, respectively;
FIG. 3a is a schematic diagram of the sensing mechanism of an organic electrochemical transistor sensor;
FIG. 3b is I of an organic electrochemical transistor sensorDS-t result graph;
FIG. 3c is a graph of survivin protein concentration as a result of changes in drain-source current for an organic electrochemical transistor sensor;
FIG. 3d is a graph of the results of conventional differential pulse voltammetry analysis of an organic electrochemical transistor sensor;
FIG. 3e is a graph of the DPV measurement of survivin concentration for the organic electrochemical transistor sensor;
wherein each error bar in c and e represents three replicate assays (n-3, standard deviation);
FIGS. 4a and 4b are graphs of the results of selectivity testing of organic electrochemical transistor sensors;
FIG. 4c is a graph of the results of a repeatability test of an organic electrochemical transistor sensor;
FIG. 4d is a graph of the results of stability testing of an organic electrochemical transistor sensor;
FIG. 5a is a calibration chart of a commercial ELISA kit for survivin protein;
FIG. 5b is a graph of organic electrochemical transistor sensor differences for 22 clinical biological samples;
fig. 5c is a graph of the correlation results of the organic electrochemical transistor sensor with a commercial ELISA kit.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
It should be noted that the embodiments and features of the embodiments may be combined with each other without conflict.
The invention is further described with reference to the following drawings and specific examples, which are not intended to be limiting.
Example 1
The embodiment provides a preparation method of an organic electrochemical transistor sensor, which comprises the following steps:
s1, preparing two-dimensional layered transition metal carbide: 0.99g LiF was dissolved in 20mL HCl under magnetic stirring and 1.25g Ti was added slowly (10 min)3AlC2Stirring for 10 hours at 40 ℃, repeatedly washing for 6 times by using deionized water until the pH value reaches 6.4, drying for 12 hours in a vacuum oven (45 ℃), and freeze-drying for 12 hours to obtain Ti3C2Tx MXene；
S2, as shown in fig. 1a, preparing a nanocomposite: ti prepared in step S13C2Mixing Tx MXene with PEDOT (PSS) in a ratio of 1:2-2:1 to obtain MXene/PEDOT (PSS);
s3, preparing an organic electrochemical transistor sensor: natural oxide p+Washing a silicon substrate (size: 1.5cm multiplied by 1cm) with acetone, isopropanol and water in sequence, drying, patterning by photoresist S1813(Shipley) and a shadow mask, and depositing patterned Cr/Au source and drain electrodes (thickness: 5nm/45nm) on p through channel structure thermal evaporation+On a silicon substrate, the working pressure of Cr thermal evaporation deposition is 1.2Pa, and the thickness is 10 nm; the working pressure of Au thermal evaporation deposition is 0.5Pa, and the thickness is 50 nm; after deposition is finished, cleaning with deionized water, drying with a nitrogen gun, and baking at 100-;
s4, modifying the organic electrochemical transistor sensor: as shown in FIG. 1b, the surface of the organic electrochemical transistor sensor prepared in step S3 is coated with MXene/PEDOT: PSS prepared in step S2, and the gate electrode is modified with survivin antibody.
The characterization parameters of the prepared organic electrochemical transistor sensor are shown in figures 1 c-g.
Example 2
To verify the functionalization process of the gate electrode in an organic electrochemical transistor sensor as described in example 1, this example characterizes CV, EIS, contact angle, and AFM.
As shown in fig. 2, this example performed an electrochemical impedance spectroscopy to verify the layer-by-layer process; EIS shows that Ret in the original state is 347.5 omega, and EIS values of Au/MAA/anti-survivin and Au/MAA/anti-survivin/BSA are 359.2 omega and 385.2 omega respectively; in CV analysis, the bare gold electrode showed a redox current of 78.2 μ A at 293mV (vs Ag/AgCl); in contrast, the response currents of Au/MAA/anti-survivin and Au/MAA/anti-survivin/BSA were reduced to 74.3. mu.A and 72.3. mu.A, respectively; at the same time, at 10mM K3[Fe(CN)6]/K4[Fe(CN)6]Scanning Rate dependence of the Mixed solution in the study, the square root (V) of the electrochemical Current and the scanning Rate1/2) Shows a linear character, indicating that the surface controlled process has iredox＝8.28×V1/2-a linear relationship of 23.57 (correlation coefficient 0.9985); this demonstrates the excellent performance of the gate electrode in the efficient transport of electrons.
Example 3
In order to improve the sensing performance of the organic electrochemical transistor sensor as described in embodiment 1, the present embodiment optimizes important factors including: pH, incubation time, temperature and concentration of the survivin antibody, and the optimized parameters are as follows: the pH was 7.0, the incubation time was 30 minutes, the temperature was 37 ℃ and the survivin antibody protein concentration was 10. mu.g/mL.
As shown in FIG. 3a, the organic electrochemical transistor sensor responds to channel current with various concentrations of survivin protein added, and the higher the concentration of protein solution used to capture the gate electrode, the change in channel current observed (I)DS) The lower the concentration of the targeted survivin protein is, the monotonic relationship exists between the concentration of the targeted survivin protein and the change of the channel current change is shown; as shown in FIG. 3b, the signal-to-noise ratio (S/N) > 3, satisfies the detection limit (10)-2ng/mL), on the contrary, the channel current is slightly changed under the condition of no survivin antibody protein functionalization, thereby illustrating the working mechanism of OECT; as shown in FIG. 3c, the proposed organic electrochemical transistor sensor achieves sensitive quantitation of survivin, an ultra-wide linear range 101-102And 10-5×105ng·mL-1Linear corresponding to the regression curve as DeltaIDS(μA)＝29.98×log[concentrationsurvivin]+115.38(R20.96) and Δ IDS(μA)＝42.78×log[concentrationsurvivin]-19.20(R20.97). As shown in fig. 3d-e, the survivin antibody modified gate electrode was also used as a working electrode in a conventional Differential Pulse Voltammetry (DPV) assay, with a limit of detection (LOD) of 100pg/mL (S/N-3) and a sensitivity of 35.3 μ a (ng · mL)-1)-1cm-2。
Example 4
To evaluate the selectivity of the organic electrochemical transistor sensor for the survivin protein assay as described in example 1, this example performed a measurement by adding an interfering molecule.
Typical high molecular weight proteins (CD63, PSA, CD44), low molecular weight molecules (glucose, tyrosine, lysine) were selected as interferents. As shown in FIGS. 4a-b, IDSThe curve at 1 ng.mL-1survivin (14.01 + -0.4 muA) induction showed high response of varying current. Correspondingly, 10ng/mL CD44(0.31 +/-0.31), PSA (0.33 +/-0.27), CD63(0.33 +/-0.27), glucose (0.42 +/-0.03), lysine (0.33 +/-0.27) and tyrosine (0.45 +/-0.29) show low response; at the same time, five sensors were tested to determine 1ng mL-1Δ in survivinCurrent responseTo evaluate reproducibility. RSD of 2.33% (fig. 4c) demonstrates the high reproducibility of the OECT sensor. In the stability monitoring (FIG. 4d), I after 1 week (443.3. + -. 8.12. mu.A), 2 weeks (440.1. + -. 8.24. mu.A), 1 month (438.1. + -. 8.14. mu.A) and 2 months (436.5. + -. 8.22. mu.A) was monitored compared to the original reaction (445. + -. 8.2. mu.A)DSIs negligible in change, IDSDescend<4％。
Example 5
This example uses an organic electrochemical transistor sensor as described in example 1 to detect survivin protein in clinical biological samples of serum.
Serum samples from a series of patients were collected by standard protocols and tested for survivin protein values by commercial ELISA kits, the calibration chart being shown in figure 5 a. The positive and negative samples are distinguished as shown in FIG. 5b (N)positive＝11，Nnegative＝11，p<0.01). As shown in FIG. 5cIt is shown that the serum survivin concentration is linearly related between the measurement of the organic electrochemical transistor sensor and the measurement of the ELISA (Y ═ 1.0015 × X +0.0039, R20.9717), indicating that there is no significant difference between the organic electrochemical transistor sensor and the commercial ELISA kit.
In conclusion, the organic electrochemical transistor sensor can be used for detecting survivin with high sensitivity.
While the invention has been described with reference to a preferred embodiment, it will be understood by those skilled in the art that various changes in form and detail may be made therein without departing from the spirit and scope of the invention.
Claims (8)
1. A method of making an organic electrochemical transistor sensor, comprising the steps of:
s1, preparing two-dimensional layered transition metal carbide: selective etching of Ti in aqueous LiF/HCl3AlC2The aluminum layer in (1) to obtain Ti3C2Tx MXene；
S2, preparing the nano composite material: ti prepared in step S13C2Mixing Tx MXene with PEDOT (PSS) in a ratio of 1:2-2:1 to obtain MXene/PEDOT (PSS);
s3, preparing an organic electrochemical transistor sensor: natural oxide p+Washing the silicon substrate with acetone, isopropanol and water in sequence, drying, and patterning with a photoresist and a shadow mask to obtain the organic electrochemical transistor sensor;
s4, modifying the organic electrochemical transistor sensor: the organic electrochemical transistor sensor prepared in the step S3 is coated with MXene/PEDOT: PSS prepared in the step S2 on its surface, and a survivin antibody is modified on the gate electrode.
2. The method according to claim 1, wherein step S1 includes:
LiF was dissolved in HCl under magnetic stirring, and Ti was added slowly3AlC2Stirring at 20-60 deg.C for a period of time, repeatedly washing with deionized water for 3-9 times until pH reaches 6-7, and drying to obtain Ti3C2Tx MXene。
3. The method of claim 1, wherein the patterning in step S3 includes:
the source electrode and the drain electrode of Cr/Au are deposited on p by thermal evaporation through a channel structure+Cleaning the silicon substrate by deionized water, drying by a nitrogen gun, and baking at the temperature of 100-140 ℃ for 15-45 seconds.
4. The method according to claim 3, wherein the working pressure for thermal evaporation deposition of Cr is 1.2Pa and the thickness is 10 nm.
5. The method according to claim 3, wherein the working pressure for thermal evaporation deposition of Au is 0.5Pa and the thickness is 50 nm.
6. The method of claim 1, wherein the parameters for modifying the survivin antibody in step S4 include: the pH value is 7.0; the time is 30 minutes; the temperature is 37 ℃; the concentration of survivin antibody protein was 10. mu.g/mL.
7. An organic electrochemical transistor sensor manufactured by the manufacturing method according to any one of claims 1 to 6.
8. Use of the organic electrochemical transistor sensor of claim 7 for highly sensitive detection of survivin.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20200096475A1 (en) * | 2017-05-09 | 2020-03-26 | Alma Mater Studiorum - Universita' Di Bologna | ORGANIC ELECTROCHEMICAL TRANSISTOR BASED ON CONDUCTIVE POLYMER AND Ag, AgX (X = Cl, I, Br) HALIDE NANOPARTICLES, AS CHEMICAL SENSOR |
| CN112578011A (en) * | 2020-12-08 | 2021-03-30 | 中国科学院苏州生物医学工程技术研究所 | Sensor and detection method for detecting dopamine and uric acid |
| US20210102900A1 (en) * | 2018-04-20 | 2021-04-08 | Qingdao University | Biosensor based on trititanium dicarbide two-dimensional metal carbide catalyzed luminol electrogenerated chemiluminescence probe and preparation method |
| CN112924505A (en) * | 2021-02-04 | 2021-06-08 | 大连理工大学 | Preparation method and application of intelligent wearable heavy metal sensing system based on bismuth film |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20200096475A1 (en) * | 2017-05-09 | 2020-03-26 | Alma Mater Studiorum - Universita' Di Bologna | ORGANIC ELECTROCHEMICAL TRANSISTOR BASED ON CONDUCTIVE POLYMER AND Ag, AgX (X = Cl, I, Br) HALIDE NANOPARTICLES, AS CHEMICAL SENSOR |
| US20210102900A1 (en) * | 2018-04-20 | 2021-04-08 | Qingdao University | Biosensor based on trititanium dicarbide two-dimensional metal carbide catalyzed luminol electrogenerated chemiluminescence probe and preparation method |
| CN112578011A (en) * | 2020-12-08 | 2021-03-30 | 中国科学院苏州生物医学工程技术研究所 | Sensor and detection method for detecting dopamine and uric acid |
| CN112924505A (en) * | 2021-02-04 | 2021-06-08 | 大连理工大学 | Preparation method and application of intelligent wearable heavy metal sensing system based on bismuth film |
Non-Patent Citations (1)
| Title |
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| XU PING等: "Combination of ultrathin micro-patterned MXene and PEDOT:Poly(styrenesulfonate) enables organic electrochemical transistor for amperometric determination of survivin protein in children osteosarcoma", MICROCHIMICA ACTA, vol. 188, pages 6 * |
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