Electric field-induced controllable molecular assembly biosensor, preparation method and application thereof

Abstract

The application provides an electric field-induced controllable molecular assembly biosensor, a preparation method and application thereof. The biosensor of the application comprises probe molecules which are orderly and orientally distributed on an interface. The biosensor of the application can stably work in a liquid phase environment; and specific interaction between the probe molecules and target substances provides an effective method for sensitive identification of disease biomarkers and environmental hazardous substances. The application realizes high sensitivity, rapid response and reliable signal analysis of biomarkers by applying an external electric field to directionally induce semiconductor materials and probe molecules to assemble a composite semiconductor layer containing the probe molecules.

Description

Technical Field

[0001] This invention belongs to the field of bioelectronic sensors, specifically relating to a biosensor with controllable molecular assembly induced by an electric field, its preparation method, and its application. Background Technology

[0002] Sensors can convert the signal to be measured into an electrical signal and are widely used in fields such as health diagnosis, drug screening, food safety and environmental monitoring. With the development of materials and solution processing technology, organic electronic devices such as organic transistors have been extensively studied in the field of sensing ((1) E. Macchia et al., Nat. Commun. 2018, 9, 3223. (2) H. Liu et al., Adv. Sci. 2024, 11, 2305347. (3) D. Ye et al., Adv. Mater. 2021, 33, 2100489.).

[0003] Organic electrochemical transistors (OETTs) possess both excellent signal conversion and amplification capabilities, making them an effective carrier for constructing biosensors. OETTs have advantages such as a wide range of material sources, high intrinsic flexibility, good biocompatibility, and the ability to be fabricated in large areas using solution methods. They have shown great application potential in the field of biosensing and have been widely used in health fields such as biological detection, early disease diagnosis, and drug development ((1) K. Guo et al., Nat. Biomed. Eng. 2021, 5, 666. (2) H. Liu et al., Sci. Adv. 2021, 7, eabg8387. (3) H. Liu et al., Adv. Sci. 2024, 11, 2305347.).

[0004] Biomarkers serve as biochemical indicators of structural or functional changes in human systems, organs, tissues, and cells. Monitoring biomarker content and conformation can be used for disease diagnosis and early warning, drug efficacy evaluation, and assessment of new therapies. However, because biomarkers in the human body are often present in low concentrations and exist in complex solution environments such as blood and tissue fluid, their detection is easily affected by interference from multiple ions, molecules, pH, and other signals, resulting in low sensitivity and poor selectivity. Therefore, there is a need to develop biosensors that can operate stably in liquid environments and achieve high sensitivity, high selectivity, and rapid response. Summary of the Invention

[0005] To overcome the shortcomings of existing technologies, one object of the present invention is to provide a biosensor comprising probe molecules ordered and oriented at an interface, which possesses numerous advantages such as high sensitivity, high selectivity, and rapid response. Another object of the present invention is to provide a method for preparing the biosensor, which utilizes solution processing technology to deposit an organic semiconductor thin film, and during the film deposition process, applies an external electric field to induce the controlled assembly of the semiconductor layer and / or probe molecules.

[0006] The technical solution of the present invention is as follows:

[0007] A biosensor comprising probe molecules with an ordered orientation distributed on an interface.

[0008] According to the embodiments of this aspect, the probe molecule is capable of specifically binding to a biomarker.

[0009] According to an embodiment of the present invention, the probe molecules are orderly oriented and distributed on at least a portion of the interface of the semiconductor layer in the biosensor.

[0010] According to an embodiment of the present invention, the semiconductor layer is an organic semiconductor layer.

[0011] According to the embodiments of this aspect, the probe molecules are controllably assembled and orderly oriented on the interface by an applied electric field.

[0012] According to an embodiment of the present invention, the semiconductor layer is assembled in a controlled manner by direction-induced external electric field.

[0013] According to an embodiment of the present invention, the semiconductor layer is first deposited using solution processing technology to form an organic semiconductor thin film, and then formed by controlled assembly oriented by applying an external electric field during the thin film deposition process.

[0014] According to an embodiment of the present invention, the semiconductor layer and the probe molecules are simultaneously and controllably assembled by an applied electric field in a directional induction process.

[0015] According to an embodiment of the present invention, an organic semiconductor thin film is first deposited using solution processing technology, and then, during the thin film deposition process, an external electric field is applied to induce orientation to form the semiconductor layer and probe molecules with ordered orientation distribution on at least a portion of the interface of the semiconductor layer.

[0016] According to an embodiment of the present invention, the biosensor includes a substrate, an electrode layer, a semiconductor layer, and an electrolyte layer; the semiconductor layer is located between the substrate and the electrolyte layer; the probe molecules are ordered and oriented at the interface of at least a portion of the semiconductor layer, and the probe molecules are capable of specifically binding to biomarkers.

[0017] According to an embodiment of the present invention, the biomarker is a disease marker, such as a disease marker derived from bodily fluids. Preferably, the biomarker is at least one of a cancer marker, an inflammation marker, a diabetes marker, and a kidney disease marker.

[0018] According to an embodiment of the present invention, the body fluid includes at least one of blood, plasma, serum, lymph, urine, saliva, sweat, and tears.

[0019] According to an embodiment of the present invention, the probe molecule is selected from at least one of enzymes, antibodies, nucleic acid aptamers, and drug molecules.

[0020] Preferably, the drug molecule is selected from at least one of natural drug molecules, chemically synthesized drug molecules, genetically engineered drug molecules, and drug-active precursor molecules. For example, the drug molecule is selected from at least one of gefitinib, osimertinib, captopril, afatinib, edalisib, rosuvastatin, and pravastatin.

[0021] For example, the probe molecule is rosuvastatin calcium, and the biomarker is C-reactive protein (CRP).

[0022] According to an embodiment of the present invention, the semiconductor layer comprises a semiconductor material. Preferably, the semiconductor material is selected from at least one or a mixture of two or more semiconductors such as polythiophene semiconductors, pyrrolopyrrole semiconductors, or naphthalimide semiconductors. Exemplarily, the semiconductor layer comprises at least one of PEDOT:PSS, PBTTT, P3HT, DPP2T-TT, P(g2T-TT), and PgBTTT.

[0023] According to an embodiment of the present invention, the semiconductor layer has at least one of a thin film structure, a bulk structure, a sheet structure, etc.

[0024] According to an embodiment of the present invention, the thickness of the semiconductor layer is 10nm-1000nm, preferably 10nm-200nm, more preferably 20nm-100nm, for example 40nm.

[0025] According to an embodiment of the present invention, the substrate may be a rigid substrate and / or a flexible substrate.

[0026] Preferably, the rigid substrate is at least one selected from glass, ceramic, plexiglass, and plastic. For example, the substrate is Corning glass.

[0027] Preferably, the flexible substrate is at least one of polyvinyl alcohol (PVA), polyester (PET), polyimide (PI), polyethylene naphthalate (PEN), paper, textile materials, silicone rubber, polylactic acid, polyurethane, etc.

[0028] According to an embodiment of the present invention, the electrode layer includes a source electrode layer, a drain electrode layer, and a gate electrode layer arranged sequentially at intervals.

[0029] Preferably, the lower surface of the source electrode layer is in contact with the substrate, the upper surface is in contact with the semiconductor layer, one side of the surface near the drain electrode layer is in contact with the semiconductor layer, and the other side of the surface is in contact with the electrolyte layer; the lower surface of the drain electrode layer is in contact with the substrate, the upper surface is in contact with the semiconductor layer, one side of the surface near the source electrode layer is in contact with the semiconductor layer, and the other side of the surface is in contact with the electrolyte layer; furthermore, the semiconductor layer between the source electrode layer and the drain electrode layer is integral.

[0030] Preferably, the lower surface of the gate electrode layer is in contact with the substrate, and the upper surface is in contact with the electrolyte layer.

[0031] Preferably, the probe molecules are oriented in an orderly manner on the interface of the gate electrode layer on the side opposite to the substrate.

[0032] According to an embodiment of the present invention, the electrode layer includes a source electrode layer, a drain electrode layer, and a gate electrode layer. The source electrode layer and the drain electrode layer are disposed on a substrate at intervals, and an electrolyte layer is disposed between the gate electrode layer and the substrate on which the source electrode layer and the drain electrode layer are disposed. Preferably, the lower surface of the source electrode layer is in contact with the substrate, the upper surface is in contact with the semiconductor layer, one side of the surface near the drain electrode layer is in contact with the semiconductor layer, and the other side of the surface is in contact with the electrolyte layer; the lower surface of the drain electrode layer is in contact with the substrate, the upper surface is in contact with the semiconductor layer, one side of the surface near the source electrode layer is in contact with the semiconductor layer, and the other side of the surface is in contact with the electrolyte layer; further, the semiconductor layer between the source electrode layer and the drain electrode layer is integral. Further, the gate electrode layer is disposed above the source electrode layer and the drain electrode layer, immersed in the electrolyte layer, and not in contact with the semiconductor layer.

[0033] Preferably, the probe molecules are arranged in an orderly orientation on the interface of the semiconductor layer on the side opposite to the substrate.

[0034] According to an embodiment of the present invention, the source electrode layer and the drain electrode layer may be parallel plate electrodes or differential finger electrodes. Further, the distance between the source electrode layer and the drain electrode layer is 10 nm-1000 μm, preferably 10-100 nm, for example, 20 nm, 30 nm, 40 nm, or 50 nm. Preferably, the thickness of the source electrode layer and the drain electrode layer may be the same or different, and are independently selected from 10-1000 nm, preferably 10-100 nm, for example, 20 nm, 30 nm, 40 nm, or 50 nm.

[0035] According to an embodiment of the present invention, the thickness of the gate electrode layer is 10-1000 nm, preferably 10-100 nm, for example 20 nm, 30 nm, 40 nm, or 50 nm.

[0036] According to an embodiment of the present invention, the shape of the electrode layer may be selected from those known in the art, such as a needle electrode with a thickness of 0.1-5 mm, preferably 0.2-2 mm, for example 0.5 mm.

[0037] According to an embodiment of the present invention, the source electrode layer, drain electrode layer and gate electrode layer are made of the same or different materials, and are independently selected from at least one of metal, alloy, metal oxide, electrochemical electrode, heavily doped semiconductor and conductive polymer.

[0038] Preferably, the metal includes at least one of gold, silver, aluminum, titanium, copper, tin, or aluminum.

[0039] Preferably, the alloy comprises at least one of a magnesium-silver alloy, a platinum alloy, or a nickel-zinc alloy.

[0040] Preferably, the metal oxide includes at least one of indium tin oxide, manganese dioxide, or lead dioxide.

[0041] Preferably, the electrochemical electrode includes silver / silver chloride, platinum-carbon, or carbon black electrodes.

[0042] Preferably, the heavily doped semiconductor comprises inorganically doped silicon, wherein the doping element is selected from phosphorus, boron, and arsenic. Exemplarily, the heavily doped semiconductor is, for example, phosphorus-doped silicon, boron-doped silicon, or arsenic-doped silicon. Further, the mass percentage of the doping element is 0.1%-3%, preferably 0.5%-2%, and for example, 1%-1.5%.

[0043] Preferably, the conductive polymer includes at least one of polyaniline, polypyrrole, and polythiophene. Preferably, the number average molecular weight of the conductive polymer is 100-1,000,000, more preferably 100-10,000, and for example, 100-10,000.

[0044] Preferably, the number average molecular weight of the polyaniline is 450-1,000,000, for example, 20,000.

[0045] Preferably, the number average molecular weight of the polypyrrole is 300-1,000,000, for example, 20,000.

[0046] Preferably, the polythiophene has the molecular formula [C4SR1R2]. n R1 and R2 are independently selected from C, O, N or H, n is a positive integer, and the number average molecular weight of the polythiophene is 400-1,000,000, for example 20,000.

[0047] According to an embodiment of the present invention, the electrolyte layer comprises at least one of an ionic solution, an ionic liquid, and an ionic organic composite electrolyte. Preferably, the ionic solution is selected from at least one of PBS buffer, HEPES buffer, and physiological saline. Preferably, the ionic liquid is selected from ionic liquids known in the art, such as [EMIM] and [TFSI]. Preferably, the ionic organic composite electrolyte comprises an ionic liquid and a polymer, wherein the mass content of the ionic liquid is 10-50 wt%, for example, 50 wt%. Further, the polymer is selected from polymethyl methacrylate (PMMA), poly(vinylidene fluoride-co-hexafluoropropylene) (PVDF-HFP), and polyethylene oxide (PEO).

[0048] For example, the ion-containing organic composite electrolyte is selected from [EMIM][TFSI] / PMMA (methylimidazolium bis(trifluoromethanesulfonyl)imide salt and polymethyl methacrylate).

[0049] According to an embodiment of the present invention, the thickness of the electrolyte layer can be 1-100 μm, preferably 1-50 μm, for example 5 μm.

[0050] The present invention also provides a method for preparing the above-mentioned biosensor, which includes at least the step of forming probe molecules. The step specifically includes: preparing a solution containing probe molecules, and under the orientation induction of an applied electric field, the probe molecules are controllably assembled and orderly oriented and distributed on the interface of the biosensor.

[0051] According to an embodiment of the present invention, the method further includes a step of forming a semiconductor layer, which specifically includes: preparing a solution containing semiconductor material, depositing a semiconductor thin film using solution processing technology, and arranging the thin film under the directional induction of an applied electric field to form the semiconductor layer.

[0052] According to an embodiment of the present invention, the formation of probe molecules and the formation of a semiconductor layer can be carried out simultaneously, specifically including: preparing a solution containing probe molecules and semiconductor materials, depositing a semiconductor thin film using solution processing technology, and forming a semiconductor layer and probe molecules with ordered orientation distribution on at least a portion of the interface of the semiconductor layer under the directional induction of an applied electric field.

[0053] According to an embodiment of the present invention, the solution processing technology can be performed using methods known in the art, such as spin coating, drop coating, shear stretching, vapor deposition, or roll-to-roll printing.

[0054] According to an embodiment of the present invention, the applied electric field refers to the application of a constant voltage, which can be selected from conditions known in the art, such as applying a constant voltage of 30V.

[0055] According to an embodiment of the present invention, the mass ratio of semiconductor material to probe molecules is not specifically limited, as long as the probe molecules can be distributed on the surface of the composite semiconductor layer.

[0056] According to embodiments of the present invention, the solvents in the solution containing the probe molecules and the solution containing the semiconductor material are the same or different, and are independently selected from solvents capable of dissolving the probe molecules and / or the semiconductor material; further, the solvent is, for example, at least one of chlorobenzene, chloroform, and o-dichlorobenzene.

[0057] According to an embodiment of the present invention, the preparation method further includes the steps of forming an electrode layer and an electrolyte layer. Specifically, the preparation method includes: forming a source electrode layer and a drain electrode layer on a substrate surface, respectively, followed by forming a semiconductor layer and probe molecules that are ordered and oriented at least partially distributed on the interface of the semiconductor layer, forming an electrolyte layer and a gate electrode layer, thereby obtaining the temperature sensor.

[0058] Alternatively, a source electrode layer, a drain electrode layer, and a gate electrode layer are disposed on the same surface of a substrate, and then semiconductor layers are disposed on the source electrode layer and the drain electrode layer, respectively. A semiconductor layer and probe molecules with ordered orientations distributed on at least a portion of the interface of the semiconductor layer are formed on the gate electrode layer side, and then an electrolyte layer is disposed to obtain the biosensor.

[0059] According to an embodiment of the present invention, the substrate can be cleaned using known methods.

[0060] According to an embodiment of the present invention, the method for forming a source electrode layer and a drain electrode layer on a substrate surface specifically involves preparing the source electrode layer and the drain electrode layer on the substrate by at least one of the following methods: vacuum phase deposition, printing, magnetron sputtering, or metal sheet bonding, such as vacuum thermal deposition. Preferably, the vacuum thermal deposition method can be performed under conditions known in the art, as long as the source electrode layer and the drain electrode layer can be obtained.

[0061] According to an embodiment of the present invention, the electrolyte layer can be obtained by at least one of the following methods: photocoupling, spin coating / drop coating, casting, spraying, or 3D printing, etc.

[0062] According to an embodiment of the present invention, the gate electrode layer can be prepared by the following methods: lamination (e.g., laminating flexible metal electrodes), shear stretching, spin coating (e.g., spin coating semiconductors), deposition, spraying (e.g., spraying inorganic non-metallic conductive materials), vapor deposition (e.g., vapor deposition of metals or alloys), and 3D printing.

[0063] According to an embodiment of the present invention, the semiconductor layer and / or electrolyte layer may further undergo annealing. Preferably, the annealing conditions are: annealing at or above 50°C for more than 1 hour, for example, annealing at 50-100°C for 1-5 hours.

[0064] The present invention also provides applications of the above-described biosensor, such as for identifying biomarkers.

[0065] Beneficial effects

[0066] The biosensor of this invention can operate stably in a liquid environment; utilizing the specific interaction between probe molecules and target substances, it provides an effective method for the sensitive identification of disease biomarkers and environmental hazards. This invention achieves high sensitivity, rapid response, and reliable signal analysis for biomarkers by applying an external electric field to directionally induce the assembly of semiconductor materials and probe molecules into a composite semiconductor layer containing probe molecules.

[0067] 1. The biosensor of the present invention is universal and has no specific requirements on the type and type of inorganic active layer material, organic active layer material, and organic-inorganic hybrid material; at the same time, various types of flexible, elastic materials and wearable materials can be used to prepare this type of sensor.

[0068] 2. Based on patterning and integration technology, this invention can prepare arrayed multifunctional sensing matrices, realize the functional modification of multiple probe molecules and biomolecule detection, and provide a universal biomolecule detection platform and structure-activity relationship research method.

[0069] 3. By screening low-cost materials, this invention enables the large-scale fabrication of flexible devices for the integration of highly sensitive biomolecular recognition and detection sensors with functional modifications of drug molecules.

[0070] 4. This invention enables the application of flexible wearable bioelectronic devices in fields such as real-time detection of biomarkers, drug screening and exploration of drug efficacy, health monitoring and disease diagnosis. Attached Figure Description

[0071] Figure 1 This is a schematic diagram of a biosensor structure for electric field-induced controllable molecular assembly; where a is a schematic diagram of an organic electrochemical transistor structure with the probe on the side gate, and b is a schematic diagram of an organic electrochemical transistor structure with the probe on the organic semiconductor layer of the channel.

[0072] Figure 2 This is a schematic diagram of the shearing film stretching device (a) used in this invention and the preparation under an applied electric field (b); an applied electric field is constructed during the solution processing by applying an applied voltage between a silicon doctor blade connected to the pressure point probe in the upper right and an aluminum substrate in the lower right; the device for applying the applied voltage can be, but is not limited to, various electrical power meters, such as B2902A;

[0073] Figure 3 The molecular structural formula of the material used in this invention;

[0074] Figure 4 The diagram shows the current change of the biosensor prepared in Example 1, where a is the output curve of the biosensor prepared in Example 1; and b is the transfer curve of the biosensor prepared in Example 1.

[0075] Figure 5 The biosensor prepared in Example 1 senses the changes in the analyte CRP. In this figure, a is the transfer curve of the biosensor prepared in Example 1 for different concentrations of CRP (the concentration of CRP gradually increases along the direction of the arrow in the figure); b is the sensing response value (voltage offset value) extracted at different concentrations of CRP.

[0076] Figure 6 The image shows the sensing changes of the biosensor prepared in Example 2 for the analyte CRP. In the image, a is the transfer curve of the biosensor prepared in Example 2 for different concentrations of CRP (the concentration of CRP gradually increases along the direction of the arrow in the figure); b is the sensing response value (voltage offset value) extracted at different concentrations of CRP.

[0077] Figure 7 This is a bar graph showing the selective recognition response of the biosensor obtained in Example 1 of the present invention to different substances. Detailed Implementation

[0078] The technical solution of the present invention will be further described in detail below with reference to specific embodiments. It should be understood that the following embodiments are merely illustrative and explanatory of the present invention, and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are covered within the scope of protection intended by the present invention.

[0079] Unless otherwise stated, the raw materials and reagents used in the following examples are commercially available products or can be prepared by known methods.

[0080] Example 1

[0081] Semiconductor layers are prepared by solution deposition, such as... Figure 1 The specific steps for the biosensor with the structure shown in Figure a are as follows:

[0082] 1) Prepare a PEDOT:PSS solution doped with 5% (v / v) ethylene glycol (PEDOT is poly(3,4-ethylenedioxythiophene), stirred at room temperature for 1 h, wherein PSS is polystyrene sulfonate, and PEDOT:PSS is a conductive polymer used as a semiconductor active material for devices). The structure of PEDOT:PSS is shown below. Figure 3 The PEDOT:PSS model is PH1000, the mass ratio of PEDOT to PSS is 1:2.5, and the number average molecular weight of PEDOT is 50,000.

[0083] 2) A 1mm thick glass slide (glass substrate) is sequentially subjected to ultrasonic treatment with secondary water, ethanol, and acetone, rinsed, and dried with nitrogen, and then subjected to a vacuum of 5×10⁻⁶. -6 Under the condition of Pa At a speed of [unclear], metallic chromium is deposited on the upper surface of the glass slide to a thickness of 5 nm, and then [unclear]... Gold is deposited onto metallic chromium at a speed of 25 nm to form three parallel chromium-gold electrodes, which are the source electrode, drain electrode, and gate electrode, respectively. Specifically, as shown below... Figure 1 As shown in Figure a, the spacing between the three electrodes is 30 nm; the thickness of the chromium-gold electrode is 30 nm, resulting in a substrate with three chromium-gold electrodes.

[0084] 3) Dissolve the rosuvastatin calcium drug molecule in the PEDOT:PSS solution obtained in step (1) to prepare a PEDOT:PSS solution containing 1 mg / mL rosuvastatin calcium.

[0085] 4) Oxygen plasma treatment step 2) After evaporating the chromium-gold electrode, the bottom surface of the glass substrate is covered with the source and drain electrodes, and then... Figure 2Using a shear-stretching film apparatus, the PEDOT:PSS solution containing drug probe molecules obtained in step 3) is deposited on the surface of the gate electrode. A constant voltage of 30V is applied during the deposition process, followed by thermal annealing at 120°C for 20 minutes to obtain a composite semiconductor layer containing probe molecules (see details). Figure 2 (See diagram b for the fabrication process under an applied electric field).

[0086] 5) The semiconductor layer material PEDOT:PSS solution obtained in step 1) is deposited on the source electrode and drain electrode by spin coating, and then thermally annealed at 120°C for 20 minutes to obtain an organic semiconductor layer with a thickness of 60 nm.

[0087] 6) Preparation of electrolyte layer: A 20mm² area... 2 20 μl of 1×PBS buffer solution was added as an electrolyte layer to a 0.5 mm thick PDMS bath. The solution covered the semiconductor layers on the source / drain electrodes and the side gate electrodes.

[0088] The biosensor consists of an electrolyte layer, electrodes, a semiconductor layer containing probe molecules, and a semiconductor layer without probe molecules. The source and drain electrode layers are made of chromium and gold, and the electrode layer is 30 nm thick. The gate electrode is made of the semiconductor material PEDOT:PSS and the drug probe molecule rosuvastatin calcium, and is 40 nm thick. The organic semiconductor active layer is made of the semiconductor material PEDOT:PSS, and the active layer is 60 nm thick.

[0089] See Figure 4 As shown in Figure a, the output curve of the source-drain current is adjusted under the condition that the gate voltage changes from 1.3V to -0.3V and the source-drain voltage changes from 0V to -0.5V.

[0090] See Figure 4 As shown in Figure b, the transfer curve of the source-drain current is adjusted under the conditions that the source-drain voltage is -0.05V (the source-drain voltage refers to the voltage applied between the source electrode and the drain electrode) and the gate voltage changes from 1.6V to 0.2V.

[0091] Therefore, the biosensor obtained in this embodiment can operate stably at a low operating voltage (-0.05V) and has a large subthreshold slope range and signal amplification effect.

[0092] See Figure 5 As shown in Figure a, the biosensor prepared in this embodiment exhibits corresponding transfer curves for different concentrations of the analyte CRP, showing a larger voltage offset (ΔV) compared to the baseline curve of 1×PBS.

[0093] See Figure 5As shown in Figure b, the biosensor prepared in this embodiment can detect the analyte CRP over a wide concentration range (10). -10 -10 -4 It has a good sensing response within mg / l.

[0094] Example 2

[0095] This embodiment prepares as follows: Figure 1 The specific steps for the biosensor shown in b are as follows:

[0096] 1) Referring to step 1) of Example 1, prepare a PEDOT:PSS solution doped with 5% volume fraction of ethylene glycol (PEDOT is poly(3,4-ethylenedioxythiophene));

[0097] 2) Referring to step 2) of Example 1, two parallel chromium-gold electrodes are formed on the upper surface of a 1mm thick glass sheet, serving as the source electrode and drain electrode, respectively. Specifically, as shown in... Figure 1 As shown in Figure b, the spacing between the two electrodes is 30 nm; the thickness of the chromium-gold electrode is 30 nm, resulting in a substrate with two chromium-gold electrodes.

[0098] 3) Prepare a PEDOT:PSS solution containing 1 mg / mL rosuvastatin calcium by referring to step 3) of Example 1.

[0099] 4) After oxygen plasma treatment step 2), the bottom surface of the glass substrate after chromium-gold electrode deposition is then subjected to the PEDOT:PSS solution containing drug probe molecules obtained in step 3). Figure 2 A shear-stretching device is used to deposit electrodes on the surface of a substrate containing electrodes. A constant voltage of 30V is applied during the deposition process, followed by thermal annealing at 120°C for 20 minutes to obtain a composite semiconductor layer containing probe molecules.

[0100] 5) Preparation of the electrolyte layer: A 10 mm² area... 2 10 μl of 1×PBS buffer solution was added as an electrolyte layer to a 0.5 mm thick PDMS bath. The solution covered the semiconductor layer on the source and drain electrodes.

[0101] 6) Preparation of the gate electrode: An Ag / AgCl electrode is placed on the surface of the electrolyte layer as the gate electrode.

[0102] See Figure 6 As shown in Figure a, the biosensor prepared in this embodiment exhibits corresponding transfer curves for different concentrations of the analyte CRP, showing a larger voltage offset (ΔV) compared to the baseline curve of 1×PBS.

[0103] See Figure 6As shown in b, the biosensor prepared in this embodiment, with the drug probe rosuvastatin calcium located in the organic semiconductor layer, can also detect the analyte CRP over a wide concentration range (10). -10 -10 -4 It has a good sensing response within mg / l.

[0104] Example 3

[0105] The biosensor device obtained in Example 1 was used to test CRP (concentration of 10). -5 Selective identification and detection of g / mL), and additionally, for concentrations of 10 g / mL. -5 The following other substances were selectively identified and detected at g / mL: α1-acid glycoprotein (α1-AGP), serum albumin (BSA), hemoglobin (Hb), and transferrin (TRF). The biosensor prepared in Example 1 was used to detect systems containing the above-mentioned interfering substances and CRP, and the results are as follows: Figure 7 As shown, the biosensor modified by electric field-induced controllable molecular assembly constructed in Example 1 can achieve selective recognition and detection of CRP.

[0106] The exemplary embodiments of the present invention have been described above. However, the scope of protection of this application is not limited to the above embodiments. Any modifications, equivalent substitutions, improvements, etc., made by those skilled in the art within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A biosensor, characterized in that, The biosensor comprises probe molecules with an ordered orientation distributed on the interface.

2. The biosensor according to claim 1, characterized in that, The probe molecules are oriented and distributed in an ordered manner on the interface through controlled assembly induced by an applied electric field. Preferably, the probe molecule is capable of specifically binding to the biomarker.

3. The biosensor according to claim 1 or 2, characterized in that, The probe molecules are arranged in an orderly orientation on at least a portion of the interface of the semiconductor layer in the biosensor; Preferably, the semiconductor layer is an organic semiconductor layer.

4. The biosensor according to claim 3, characterized in that, The semiconductor layer is assembled in a controlled manner by direction induction under an applied electric field; Preferably, the semiconductor layer is formed by first depositing an organic semiconductor thin film using solution processing technology, and then by controlled assembly oriented by applying an external electric field during the thin film deposition process; Preferably, the semiconductor layer and the probe molecules are simultaneously and controllably assembled under directional induction by an applied electric field; Preferably, an organic semiconductor thin film is first deposited using solution processing technology, and then, during the film deposition process, an external electric field is applied to induce orientation to form the semiconductor layer and probe molecules with ordered orientation distribution on at least a portion of the interface of the semiconductor layer.

5. The biosensor according to any one of claims 1-4, characterized in that, The biosensor includes a substrate, an electrode layer, a semiconductor layer, and an electrolyte layer; the semiconductor layer is located between the substrate and the electrolyte layer; the probe molecules are ordered and oriented at the interface of at least a portion of the semiconductor layer, and the probe molecules are capable of specifically binding to biomarkers.

6. The biosensor according to claim 5, characterized in that, The electrode layer includes a source electrode layer, a drain electrode layer, and a gate electrode layer that are spaced apart, and the probe molecules are oriented in an orderly manner on the interface of the gate electrode layer on the side opposite to the substrate. Alternatively, the electrode layer includes a source electrode layer, a drain electrode layer, and a gate electrode layer, with the source electrode layer and drain electrode layer disposed at intervals on the substrate, and an electrolyte layer disposed between the gate electrode layer and the substrate on which the source electrode layer and drain electrode layer are disposed, and the probe molecules are orderly oriented and distributed on the interface of the semiconductor layer on the side opposite to the substrate.

7. The method for preparing the biosensor according to any one of claims 1-6, characterized in that, It includes at least the step of forming probe molecules, which specifically includes: preparing a solution containing probe molecules, and under the directional induction of an applied electric field, the probe molecules are controllably assembled and orderly oriented and distributed on the interface of the biosensor.

8. The preparation method according to claim 7, characterized in that, The preparation method further includes a step of forming a semiconductor layer, which specifically includes: preparing a solution containing semiconductor material, depositing a semiconductor thin film using solution processing technology, and assembling the thin film in a controlled manner to form the semiconductor layer under the directional induction of an applied electric field.

9. The preparation method according to claim 8, characterized in that, The simultaneous formation of probe molecules and semiconductor layer specifically includes: preparing a solution containing probe molecules and semiconductor materials; depositing a semiconductor thin film using solution processing technology; and, under the directional induction of an applied electric field, controllably assembling a semiconductor layer and probe molecules with ordered orientation distribution on at least a portion of the interface of the semiconductor layer.

10. The application of the biosensor according to any one of claims 1-6 in identifying biomarkers.

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