阅读说明：本技术 一种基于纳米复合材料的生物传感器及其构建方法 (Biosensor based on nano composite material and construction method thereof ) 是由 惠俊敏 于 2020-10-28 设计创作，主要内容包括：本发明公开了一种基于纳米复合材料的生物传感器,包括基因探针和复合材料,复合材料的表面固定连接有辅助工作基因探针,复合材料的表面固定连接有玻璃碳电极,本发明涉及生物传感器技术领域。该基于纳米复合材料的生物传感器及其构建方法,玻碳电极在打磨后进行抛光,能够将玻碳电极抛光处理的更加充分,通过对抛光后的玻碳电极进行超声清洗能够有效的提高玻碳电极自身清洁程度,提高了玻碳电极的敏感程度,使得生物传感器整体能够对低浓度的检测源进行有效结合,间接的提升了检测效率和检测的精确度,使得玻碳电极在后期安装时能够与纳米材料稳定的结合在一起,大大的缩短了生物传感器整体的制造时间,提高了生产效率。(The invention discloses a biosensor based on a nano composite material, which comprises a gene probe and a composite material, wherein the surface of the composite material is fixedly connected with an auxiliary working gene probe, and the surface of the composite material is fixedly connected with a glassy carbon electrode. This biosensor based on nano-composite and construction method thereof, glassy carbon electrode polishes after polishing, can polish the more abundant of handling glassy carbon electrode, carry out ultrasonic cleaning through the glassy carbon electrode after polishing and can effectually improve glassy carbon electrode self clean degree, the sensitive degree of glassy carbon electrode has been improved, make biosensor whole can effectively combine the detection source of low concentration, indirect promotion detection efficiency and the accuracy that detects, make glassy carbon electrode can be in the same place with the stable combination of nano-material when the later stage installation, great shortening the holistic manufacturing time of biosensor, and the production efficiency is improved.)

1. A nanocomposite-based biosensor comprising a gene probe (1) and a composite (2), characterized in that: the surface of the composite material (2) is fixedly connected with an auxiliary working gene probe (3), and the surface of the composite material is fixedly connected with a glassy carbon electrode (4).

2. A construction method of a biosensor based on a nano composite material is characterized by comprising the following steps: the method specifically comprises the following steps:

step 1: basic treatment of glassy carbon electrode surface: placing the glassy carbon electrode on the surface of an abrasive material with the diameter of 1-1.5 microns for grinding, transferring the glassy carbon electrode to the surface of the abrasive material with the diameter of 0.05-0.08 microns for polishing after grinding for 5-10 minutes;

step 2: advanced treatment of the surface of the glassy carbon electrode: transferring all the polished glassy carbon electrodes into a mixed solution of deionized water and ethanol for ultrasonic cleaning, stopping the machine for 1 minute every 5 minutes of cleaning, and taking out the glassy carbon electrodes for natural drying after 3-5 times of cleaning;

and step 3: high-order treatment of the surface of the glassy carbon electrode: scanning in electrolyte with pH value of 7 at 0.8V for 120 s, placing the glassy carbon electrode in electrolyte with pH value of 7 at 0.2-1.2V for 30-60 s, placing the glassy carbon electrode on wet flannelette, and adding a small amount of water for polishing;

and 4, step 4: preparing nano materials: placing aluminum silicate in purified water, completely stripping the aluminum silicate into a single-piece layer, gradually adding polyurethane into the mixture, allowing the aluminum silicate and the polyurethane to react naturally, removing the purified water after mixing for 5-10 minutes, allowing the layered aluminum silicate to aggregate, and clamping the polyurethane between the layered aluminum silicate to form a nano material;

and 5: basic processing of related probes: dispersing the gene probe in a hydrochloric acid diluent, dripping the gene probe to the surface of a glassy carbon electrode, then placing the glassy carbon electrode in a thermostat for incubation for 30-50 minutes, and then washing the electrode;

step 6: high-order processing of related probes: dispersing the auxiliary working gene probe in a hydrochloric acid solution, centrifugally purifying the prepared nano material, mixing 150-microliter of the nano material with 30-60 microliter of the auxiliary working gene probe, and then carrying out oscillation cultivation in an oscillator for 10-20 minutes;

and 7: assembling treatment: and gradually adding the solution after the oscillation is finished to the surfaces of the gene probe and the glassy carbon electrode solution to form the biosensor.

3. The method of claim 2, wherein the nanocomposite-based biosensor is constructed by: in the step 1, the grinding material is MgO powder with more than 200 meshes.

4. The method of claim 2, wherein the nanocomposite-based biosensor is constructed by: in the step 3, the electrolyte is a PBS solution.

5. The method of claim 2, wherein the nanocomposite-based biosensor is constructed by: in the step 3, when the glassy carbon electrode is polished, severe pollution or pockmarks exist, and scratches can be subjected to mechanical polishing treatment.

6. The method of claim 2, wherein the nanocomposite-based biosensor is constructed by: in the step 4, an auto-heating method is used when the purified water is removed.

7. The method of claim 2, wherein the nanocomposite-based biosensor is constructed by: in step 6, the device needs to be kept at a constant temperature during the shaking culture.

8. The method of claim 2, wherein the nanocomposite-based biosensor is constructed by: in the step 4, when the two are naturally reacted, the ambient temperature must be adjusted to room temperature.

Technical Field

The invention relates to the technical field of biosensors, in particular to a biosensor based on a nano composite material and a construction method thereof.

Background

A biosensor is an instrument that is sensitive to a biological substance and converts its concentration into an electrical signal for detection. Is an analysis tool or system composed of immobilized biological sensitive material as recognition element (including enzyme, antibody, antigen, microbe, cell, tissue, nucleic acid, etc.), proper physicochemical transducer (such as oxygen electrode, photosensitive tube, field effect tube, piezoelectric crystal, etc.) and signal amplification device. The biosensor has the functions of a receptor and a transducer. In designing a biosensor, it is an extremely important prerequisite to select a functional recognition substance suitable for a measurement target. Taking into account the characteristics of the resulting composite. The transducer is selected according to the chemical change or physical change caused by the sensitive element prepared by the molecular recognition functional substance, and the method is another important link for developing a high-quality biosensor. The generation or consumption of light, heat, chemical substances, etc. in the sensitive element can produce corresponding variation. Based on these variations, an appropriate transducer may be selected.

Many nanomaterials construct the biological detection platform at present and can carry out the short-term test to numerous sensitive viruses, but in actual detection, biosensor has certain requirement to the self concentration of testing source when detecting, and the self concentration base of testing source can not directly be regulated and control to traditional nanomaterial biosensor's manufacturing is very complicated, and the manufacturing time is longer.

Disclosure of Invention

Technical problem to be solved

Aiming at the defects of the prior art, the invention provides a biosensor based on a nano composite material and a construction method thereof, and solves the problems that in actual detection, the biosensor has certain requirements on the concentration of a detection source during detection, the concentration base of the detection source cannot be directly regulated and controlled, and the traditional nano material biosensor is extremely complex in manufacture and long in manufacture time.

(II) technical scheme

In order to achieve the purpose, the invention is realized by the following technical scheme: a biosensor based on a nano composite material comprises a gene probe and a composite material, wherein the surface of the composite material is fixedly connected with an auxiliary working gene probe, and the surface of the composite material is fixedly connected with a glassy carbon electrode.

The invention also discloses a construction method of the biosensor based on the nano composite material, which comprises the following steps:

step 1: basic treatment of glassy carbon electrode surface: placing the glassy carbon electrode on the surface of an abrasive material with the diameter of 1-1.5 microns for grinding, transferring the glassy carbon electrode to the surface of the abrasive material with the diameter of 0.05-0.08 microns for polishing after grinding for 5-10 minutes;

step 2: advanced treatment of the surface of the glassy carbon electrode: transferring all the polished glassy carbon electrodes into a mixed solution of deionized water and ethanol for ultrasonic cleaning, stopping the machine for 1 minute every 5 minutes of cleaning, and taking out the glassy carbon electrodes for natural drying after 3-5 times of cleaning;

and step 3: high-order treatment of the surface of the glassy carbon electrode: scanning in electrolyte with pH value of 7 at 0.8V for 120 s, placing the glassy carbon electrode in electrolyte with pH value of 7 at 0.2-1.2V for 30-60 s, placing the glassy carbon electrode on wet flannelette, and adding a small amount of water for polishing;

and 4, step 4: preparing nano materials: placing aluminum silicate in purified water, completely stripping the aluminum silicate into a single-piece layer, gradually adding polyurethane into the mixture, allowing the aluminum silicate and the polyurethane to react naturally, removing the purified water after mixing for 5-10 minutes, allowing the layered aluminum silicate to aggregate, and clamping the polyurethane between the layered aluminum silicate to form a nano material;

and 5: basic processing of related probes: dispersing the gene probe in a hydrochloric acid diluent, dripping the gene probe to the surface of a glassy carbon electrode, then placing the glassy carbon electrode in a thermostat for incubation for 30-50 minutes, and then washing the electrode;

step 6: high-order processing of related probes: dispersing the auxiliary working gene probe in a hydrochloric acid solution, centrifugally purifying the prepared nano material, mixing 150-microliter of the nano material with 30-60 microliter of the auxiliary working gene probe, and then carrying out oscillation cultivation in an oscillator for 10-20 minutes;

and 7: assembling treatment: gradually adding the solution after the oscillation is finished to the surfaces of the gene probe and the glassy carbon electrode solution to form the biosensor

Preferably, in the step 1, the grinding material is MgO powder with a mesh size of 200 mesh or more.

Preferably, in the step 3, the electrolyte is a PBS solution.

Preferably, in the step 3, when the glassy carbon electrode is ground, severe pollution or pockmarks exist, and scratches can be subjected to mechanical polishing treatment.

Preferably, in the step 4, an autothermal heating method is used for removing the purified water.

Preferably, in step 6, the device needs to be kept at a constant temperature during shaking culture.

Preferably, in the step 4, when the both are naturally reacted, the ambient temperature must be adjusted to room temperature.

(III) advantageous effects

The invention provides a biosensor based on a nano composite material and a construction method thereof. Compared with the prior art, the method has the following beneficial effects:

(1) the biosensor based on the nano composite material and the construction method thereof are characterized in that the method comprises the following steps of 1: basic treatment of glassy carbon electrode surface: placing the glassy carbon electrode on the surface of an abrasive material with the diameter of 1-1.5 microns for grinding, transferring the glassy carbon electrode to the surface of the abrasive material with the diameter of 0.05-0.08 microns for polishing after grinding for 5-10 minutes; step 2: advanced treatment of the surface of the glassy carbon electrode: the method comprises the steps of transferring all polished glassy carbon electrodes into mixed liquid of deionized water and ethanol for ultrasonic cleaning, stopping for 1 minute every 5 minutes of cleaning, taking out the glassy carbon electrodes for natural drying after 3-5 times of cleaning, and polishing the glassy carbon electrodes after polishing through the combined arrangement of the step 1 and the step 2, so that the polished glassy carbon electrodes can be more fully polished, the self cleaning degree of the glassy carbon electrodes can be effectively improved by performing ultrasonic cleaning on the polished glassy carbon electrodes, the sensitivity of the glassy carbon electrodes is improved, the whole biosensor can effectively combine low-concentration detection sources, and the detection efficiency and the detection accuracy are indirectly improved.

(2) The biosensor based on the nano composite material and the construction method thereof are characterized in that the method comprises the following steps of 3: high-order treatment of the surface of the glassy carbon electrode: scanning in electrolyte with pH value of 7 at 0.8V for 120 s, placing the glassy carbon electrode in electrolyte with pH value of 7 at 0.2-1.2V for 30-60 s, placing the glassy carbon electrode on wet flannelette, and adding a small amount of water for polishing; and 4, step 4: preparing nano materials: the aluminum silicate is placed in purified water, the aluminum silicate is completely stripped into a single-piece layer, then the polyurethane is gradually added into the mixture, the aluminum silicate and the purified water react naturally, after the aluminum silicate and the purified water are mixed for 5-10 minutes, the layered aluminum silicate is gathered, the polyurethane is clamped between the layered aluminum silicate, the nano material is formed, through the setting in the step 3, the glassy carbon electrode can be stably combined with the nano material in the later installation process, the overall manufacturing time of the biosensor is greatly shortened, the production efficiency is improved, through the setting in the step 4, the purified water is removed from the nano material in the production process, the overall concentration of the nano material can be improved, and the overall reliability of the biosensor is improved.

(3) The biosensor based on the nano composite material and the construction method thereof are characterized in that the method comprises the following steps: basic processing of related probes: dispersing the gene probe in a hydrochloric acid diluent, dripping the gene probe to the surface of a glassy carbon electrode, then placing the glassy carbon electrode in a thermostat for incubation for 30-50 minutes, and then washing the electrode; step 6: high-order processing of related probes: dispersing the auxiliary working gene probe in a hydrochloric acid solution, centrifugally purifying the prepared nano material, mixing 150-microliter of the auxiliary working gene probe with 30-60 microliter of the nano material, then carrying out oscillation cultivation in an oscillator for 10-20 minutes, and further processing the gene probe through the combined arrangement of the step 5 and the step 6, so that the gene probe can be firmly and uniformly distributed on the surface of the nano material, the later detection reliability is indirectly improved, the whole step operation is clear, the logic is clear, and the method is convenient for workers to execute.

Drawings

FIG. 1 is a schematic structural diagram of the present invention.

In the figure, 1, gene probe; 2. a composite material; 3. an auxiliary working gene probe; 4. a glassy carbon electrode.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings and the attached tables 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 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.

Referring to table 1 and fig. 1, a biosensor based on a nanocomposite includes a gene probe 1 and a composite 2, an auxiliary working gene probe 3 is fixedly connected to a surface of the composite 2, and a glassy carbon electrode 4 is fixedly connected to a surface of the composite.

The embodiment of the invention provides three technical schemes: a biosensor based on a nano composite material and a construction method thereof specifically comprise the following embodiments:

example 1

Step 1: basic treatment of glassy carbon electrode surface: placing the glassy carbon electrode on the surface of a 1-micron abrasive material for grinding, transferring the glassy carbon electrode to the surface of a 0.05-micron abrasive material for polishing after grinding for 5 minutes;

step 2: advanced treatment of the surface of the glassy carbon electrode: transferring all the polished glassy carbon electrodes into a mixed solution of deionized water and ethanol for ultrasonic cleaning, stopping the machine for 1 minute every 5 minutes of cleaning, and taking out the glassy carbon electrodes for natural drying after 3 times of cleaning;

and step 3: high-order treatment of the surface of the glassy carbon electrode: scanning in 0.8V electrolyte with pH value of 7 for 120 seconds, then placing the glassy carbon electrode in 0.2V electrolyte for cyclic scanning for 30 seconds, then placing the glassy carbon electrode on wet flannelette, and adding a small amount of water for polishing;

and 4, step 4: preparing nano materials: placing aluminum silicate in purified water, completely stripping the aluminum silicate into a single-piece layer, gradually adding polyurethane into the mixture, allowing the aluminum silicate and the polyurethane to react naturally, removing the purified water after mixing for 5 minutes, allowing the layered aluminum silicate to aggregate, and clamping the polyurethane between the layered aluminum silicate to form a nano material;

and 5: basic processing of related probes: dispersing the gene probe in a hydrochloric acid diluent, dripping the gene probe to the surface of a glassy carbon electrode, placing the glassy carbon electrode in a thermostat for incubation for 30 minutes, and then washing the electrode;

step 6: high-order processing of related probes: dispersing the auxiliary working gene probe in a hydrochloric acid solution, centrifugally purifying the prepared nano material, mixing 150 microliters of the nano material with 30 microliters of the auxiliary working gene probe, and then oscillating and culturing for 10 minutes in an oscillator;

and 7: assembling treatment: and gradually adding the solution after the oscillation is finished to the surfaces of the gene probe and the glassy carbon electrode solution to form the biosensor.

Example 2

Step 1: basic treatment of glassy carbon electrode surface: placing the glassy carbon electrode on the surface of a 1.25-micron abrasive material for grinding, and transferring the glassy carbon electrode to the surface of a 0.06-micron abrasive material for polishing after 7.5 minutes of grinding;

step 2: advanced treatment of the surface of the glassy carbon electrode: transferring all the polished glassy carbon electrodes into a mixed solution of deionized water and ethanol for ultrasonic cleaning, stopping the machine for 1 minute every 5 minutes of cleaning, and taking out the glassy carbon electrodes for natural drying after cleaning for 4 times;

and step 3: high-order treatment of the surface of the glassy carbon electrode: scanning in 0.8V electrolyte with pH value of 7 for 120 seconds, then placing the glassy carbon electrode in 0.7V electrolyte for circulating scanning for 45 seconds, then placing the glassy carbon electrode on wet flannelette, and adding a small amount of water for polishing;

and 4, step 4: preparing nano materials: placing aluminum silicate in purified water, completely stripping the aluminum silicate into a single-piece layer, gradually adding polyurethane into the mixture, allowing the aluminum silicate and the polyurethane to react naturally, removing the purified water after mixing for 7.5 minutes, allowing the layered aluminum silicate to aggregate, and clamping the polyurethane between the layered aluminum silicate to form a nano material;

and 5: basic processing of related probes: dispersing the gene probe in a hydrochloric acid diluent, dripping the gene probe to the surface of a glassy carbon electrode, placing the glassy carbon electrode in a thermostat for culturing for 40 minutes, and then washing the electrode;

step 6: high-order processing of related probes: dispersing the auxiliary working gene probe in a hydrochloric acid solution, centrifugally purifying the prepared nano material, mixing 175 microliters of the nano material with 45 microliters of the auxiliary working gene probe, and then oscillating and culturing for 15 minutes in an oscillator;

and 7: assembling treatment: and gradually adding the solution after the oscillation is finished to the surfaces of the gene probe and the glassy carbon electrode solution to form the biosensor.

Example 3

Step 1: basic treatment of glassy carbon electrode surface: placing the glassy carbon electrode on the surface of a 1.5-micron abrasive material for grinding, transferring the glassy carbon electrode to the surface of a 0.08-micron abrasive material for polishing after grinding for 10 minutes;

step 2: advanced treatment of the surface of the glassy carbon electrode: transferring all the polished glassy carbon electrodes into a mixed solution of deionized water and ethanol for ultrasonic cleaning, stopping the machine for 1 minute every 5 minutes of cleaning, and taking out the glassy carbon electrodes for natural drying after 5 times of cleaning;

and step 3: high-order treatment of the surface of the glassy carbon electrode: scanning in electrolyte with pH value of 7 at 0.8V for 120 s, placing the glassy carbon electrode in electrolyte with pH value of 7 at 1.2V, circularly scanning for 60 s, placing the glassy carbon electrode on wet flannelette, and adding a small amount of water for polishing;

and 4, step 4: preparing nano materials: placing aluminum silicate in purified water, completely stripping the aluminum silicate into a single-piece layer, gradually adding polyurethane into the mixture, allowing the aluminum silicate and the polyurethane to react naturally, removing the purified water after mixing for 10 minutes, allowing the layered aluminum silicate to aggregate, and clamping the polyurethane between the layered aluminum silicate to form a nano material;

and 5: basic processing of related probes: dispersing the gene probe in a hydrochloric acid diluent, dripping the gene probe to the surface of a glassy carbon electrode, placing the glassy carbon electrode in a thermostat for culturing for 50 minutes, and then washing the electrode;

step 6: high-order processing of related probes: dispersing the auxiliary working gene probe in a hydrochloric acid solution, centrifugally purifying the prepared nano material, mixing 200 microliters of the nano material with 60 microliters of the auxiliary working gene probe, and then oscillating and culturing for 20 minutes in an oscillator;

and 7: assembling treatment: and gradually adding the solution after the oscillation is finished to the surfaces of the gene probe and the glassy carbon electrode solution to form the biosensor.

Through the combined arrangement of the step 1 and the step 2, the glassy carbon electrode is polished after being polished, so that the polishing treatment of the glassy carbon electrode can be more sufficient, the self-cleaning degree of the glassy carbon electrode can be effectively improved through the ultrasonic cleaning of the polished glassy carbon electrode, the sensitivity of the glassy carbon electrode is improved, the whole biosensor can be effectively combined with a low-concentration detection source, the detection efficiency and the detection accuracy are indirectly improved, the glassy carbon electrode can be stably combined with a nano material during the later installation through the arrangement of the step 3, the whole manufacturing time of the biosensor is greatly shortened, the production efficiency is improved, and through the arrangement of the step 4, the pure water is removed from the nano material in the production, the whole concentration of the nano material can be improved, and the whole reliability of the biosensor is improved, through the combined setting of step 5 and step 6, through the further processing to gene probe for gene probe can be firm and even distribution on the nano-material surface, the indirect reliability that has improved later stage and detected, and whole step operation is clear, and the logic is clear, makes things convenient for the staff to carry out.

Comparative experiment

According to the three embodiments, the existing manufacturer can produce three biosensors, after the three biosensors are basically processed, the three biosensors are compared with the common biosensor in the accuracy and production time, as shown in table 1, through laboratory tests, the processing accuracy in the embodiment is 89% at least, which is 7% higher than the comparison example, the production time is 8.9 hours at the longest, which is 2% shorter than the comparison example.

Table 1: accuracy and production time of biosensor and comparative example comparison table

And those not described in detail in this specification are well within the skill of those in the art.

It is noted that, herein, relational terms such as first and second, and the like may be used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Also, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus.

Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.