CN117288816A - MXene-DNA-based composite material, sensor, preparation method of sensor and application of sensor in detection of doxorubicin - Google Patents
MXene-DNA-based composite material, sensor, preparation method of sensor and application of sensor in detection of doxorubicin Download PDFInfo
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- CN117288816A CN117288816A CN202310961750.4A CN202310961750A CN117288816A CN 117288816 A CN117288816 A CN 117288816A CN 202310961750 A CN202310961750 A CN 202310961750A CN 117288816 A CN117288816 A CN 117288816A
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- AOJJSUZBOXZQNB-TZSSRYMLSA-N Doxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-TZSSRYMLSA-N 0.000 title claims abstract description 141
- 229960004679 doxorubicin Drugs 0.000 title claims abstract description 69
- 239000002131 composite material Substances 0.000 title claims abstract description 58
- 238000002360 preparation method Methods 0.000 title claims abstract description 19
- 238000001514 detection method Methods 0.000 title claims description 22
- 244000309466 calf Species 0.000 claims abstract description 32
- 210000001541 thymus gland Anatomy 0.000 claims abstract description 32
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- 238000000034 method Methods 0.000 claims abstract description 20
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- C01B—NON-METALLIC ELEMENTS; COMPOUNDS THEREOF; METALLOIDS OR COMPOUNDS THEREOF NOT COVERED BY SUBCLASS C01C
- C01B32/00—Carbon; Compounds thereof
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- C01B32/921—Titanium carbide
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y30/00—Nanotechnology for materials or surface science, e.g. nanocomposites
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- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
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- C01B21/00—Nitrogen; Compounds thereof
- C01B21/082—Compounds containing nitrogen and non-metals and optionally metals
- C01B21/0828—Carbonitrides or oxycarbonitrides of metals, boron or silicon
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- G01N27/28—Electrolytic cell components
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Abstract
The invention discloses a composite material based on MXene-DNA, a sensor, a preparation method thereof and application thereof in detecting doxorubicin, wherein the composite material adopts two-dimensional MXene as a carrier to load calf thymus DNA, and utilizes relatively strong interaction between calf thymus DNA and doxorubicin to specifically detect the doxorubicin and display the doxorubicin in an electric signal form, so that the concentration change of the doxorubicin is accurately detected in a complex body fluid environment. The sensor has higher sensitivity, is favorable for realizing real-time detection in practical application, thereby realizing the effect of real-time drug delivery and being the best absorption state of the drug. The loading of DNA is realized through electrostatic action and hydrogen bonding action; complex reaction conditions and additional reagents are not required, so the method is simple; the preparation of the sensor coats the composite material based on the MXene-DNA on the glassy carbon electrode, the preparation method is simple, and the sensor can be immediately manufactured and used or prepared for later use and has wide application.
Description
Technical Field
The invention belongs to the technical field of medical detection, and particularly relates to an MXene-DNA (deoxyribonucleic acid) composite material, a sensor, a preparation method thereof and application thereof in detection of doxorubicin.
Background
With the increasing number of cancer and tumor cases, small molecule drugs are widely used, and it is important to be able to accurately control and monitor the use of these drugs. On the one hand, in chemotherapy, it is important to detect and maintain reasonable concentrations of anticancer drugs to prevent side effects of the drugs. On the other hand, these drugs may pose serious health risks as they may be expelled through the patient's feces and urine and may contaminate the aquatic environment. Thus DNA and small molecule drugs are an effective therapeutic approach for the treatment and prevention of human diseases. Small molecule drugs can bind to specific genes on DNA, thereby regulating gene expression. For example, small molecule drugs can bind to cytosine polymerase on DNA, blocking replication of DNA, and thus blocking proliferation of cells. In addition, small molecule drugs can also bind to transcription factors on DNA, blocking transcription of genes, and thus blocking expression of genes. In addition, small molecule drugs can also block DNA binding by binding to specific base sequences of DNA, thereby blocking DNA transcription and replication. These processes produce changes in the electrical signal.
At present, the methods for detecting small molecules by fluorescence spectrum, mass spectrum, high performance liquid chromatography and other technologies are numerous, but the methods have the problems of complicated technical process, difficult elimination, large interference, high cost and the like.
Disclosure of Invention
In order to overcome the defects in the prior art, the first aim of the invention is to provide a composite material based on MXene-DNA, wherein a two-dimensional transition metal material with conductivity is combined with DNA, so that the combination of the specificity of doxorubicin can be realized, and the specificity of an electric signal is changed.
The second object of the present invention is to provide a method for preparing the above-mentioned MXene-DNA based composite material.
A third object of the invention is to provide a sensor.
A fourth object of the invention is to provide a method for manufacturing the above sensor.
A fifth object of the invention is to provide the use of the above sensor for detecting doxorubicin.
The first object of the invention can be achieved by adopting the following technical scheme:
a composite material based on MXene-DNA comprises a two-dimensional material MXene and calf thymus DNA, wherein the calf thymus DNA is connected to the surface of the two-dimensional material MXene through electrostatic action and/or hydrogen bonding.
Further, the two-dimensional material MXene is Ti 3 C 2 、Ti 2 C、Nb 2 C、V 2 C、(Ti 0.5 Nb 0.5 ) 2 C、(V 0.5 Cr 0.5 ) 3 C 2 、Ti 3 CN or Ta 4 C 3 Any one of the following.
Further, the mass ratio of the MXene to calf thymus DNA is (100-1000): 1.
the second object of the invention can be achieved by adopting the following technical scheme:
a preparation method of a composite material based on MXene-DNA comprises the following steps:
and (3) dissolving and mixing calf thymus DNA and MXene, and then carrying out ultrasonic treatment to obtain the MXene-DNA composite material probe.
Further, the temperature during ultrasonic treatment is 10-20 ℃, and the ultrasonic treatment time is 10-30min.
The third object of the invention can be achieved by adopting the following technical scheme:
a sensor comprising an MXene-DNA based composite material of any one of the above.
Further, the MXene-DNA based composite material is coated on a glassy carbon electrode.
The fourth object of the invention can be achieved by adopting the following technical scheme:
a method of manufacturing a sensor comprising the steps of:
and (3) dripping the composite material based on the MXene-DNA on a glassy carbon electrode, and fixing by adopting a Nafion solution after air drying.
Further, the concentration of the Nafion solution is 0.5-5%; the preferred concentration of Nafion solution is 0.5%.
The fifth object of the present invention can be achieved by adopting the following technical scheme:
the application of the MXene-DNA based composite material or the sensor in the detection of the doxorubicin is realized by an electrochemical method.
Furthermore, the electrochemical method is a three-electrode system, the working electrode is the sensor, the counter electrode is a platinum wire electrode, and the reference electrode is Ag/AgCl
Compared with the prior art, the invention has the beneficial effects that:
1. according to the composite material based on the MXene-DNA, the novel two-dimensional material MXene with high conductivity and large surface area is adopted as a carrier to load calf thymus DNA with higher stability and rich sequencing data as a sensing signal probe, the calf thymus DNA and the doxorubicin have relatively strong interaction, the doxorubicin molecule is combined with the DNA and has strong specificity, and other small organic molecules have no interaction or weak interaction with the DNA, so that the doxorubicin can be specifically detected and expressed in the form of an electric signal, and the concentration change of the doxorubicin can be accurately detected in a complex body fluid environment.
2. According to the preparation method of the composite material based on the MXene-DNA, the MXene is used as a carrier, and calf thymus DNA is adsorbed on the surface of the MXene through electrostatic action and hydrogen bonding action, so that the loading of the DNA is realized; the electrostatic action and the hydrogen bonding do not need complex reaction conditions and additional reagents, so the method is simple, and the obtained MXene-DNA composite material is suitable for various scenes.
3. The sensor and the application thereof in detecting the doxorubicin have higher sensitivity, can detect the change of the electric signal in a shorter time, are favorable for realizing real-time detection in practical application, and realize the effect of real-time drug delivery, thereby being the best absorption state of the drug.
4. The preparation method of the sensor is to coat the composite material based on MXene-DNA on the glassy carbon electrode, has simple preparation method, can be immediately prepared for use or prepared for later use, and has wide application.
Drawings
FIG. 1 is a schematic illustration of a MXene-DNA composite based sensor for real-time detection of doxorubicin in complex body fluids;
FIG. 2 is an i-t curve for an MXene-DNA composite based sensor in aqueous doxorubicin solutions of varying concentrations (pH=7); wherein the concentration of doxorubicin is 0.34nM, 0.69nM, 1.12nM, 1.38nM, 1.72nM, 2.24nM, 2.75nM, 3.44nM, respectively, according to the arrow direction;
FIG. 3 is a linear relationship between doxorubicin concentration and current based on an MXene-DNA composite sensor;
FIG. 4 is an i-t pattern of detection of multiple small molecules based on an MXene-DNA composite sensor;
FIG. 5 is a bar graph of a plurality of small molecules detected by an MXene-DNA composite based sensor and their current variation;
FIG. 6 is a graph comparing current changes for doxorubicin detection based on an MXene-DNA composite sensor with an MXene alone;
FIG. 7 is a graph comparing the current change of doxorubicin detected by DNA alone.
Detailed Description
The technical scheme of the present invention will be clearly and completely described in the following in connection with specific embodiments. It will be apparent that the described embodiments are merely some, but not all, embodiments of the invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
MXene has been found to be of great interest in the biosensing field due to its ultra-thin structure and unique physicochemical properties. MXene is a new two-dimensional transition metal carbide or nitride composed of transition metal carbides, carbonitrides and nitrides, and is typically synthesized by etching a "group layer of the" MAX "phase (where" M "represents the transition metal," a "is an element of group IIIA or IVA of the periodic table, and X is C and/or N) with hydrofluoric acid (HF). Similar to graphene, MXene is a very potential adsorption material due to its excellent planar conductivity and absorptivity, and its high specific surface area, sufficient surface functional groups, and good hydrophilicity.
The invention provides a composite material based on MXene-DNA, a sensor, a preparation method thereof and application thereof in detecting doxorubicin.
A composite material based on MXene-DNA comprises a two-dimensional material MXene and calf thymus DNA, wherein the calf thymus DNA is connected to the surface of the two-dimensional material MXene through electrostatic action and/or hydrogen bonding.
The MXene is used as a carrier, and the surface of the carrier and calf thymus DNA form hydrogen bond and electrostatic action, so that calf thymus DNA is adsorbed and loaded on the MXene to form the composite material based on the MXene-DNA. When the DNA contacts the MXene surface, hydrogen bonds and hydrophobic bonds on the MXene surface will interact with the DNA reverse adenine and cytosine bases to form hydrogen bonds and hydrophobic bond bonds. This will firmly fix the DNA on the MXene surface, effectively loading the DNA. Wherein DNA can be specifically bound with drug molecules, the binding leads to a change of DNA configuration and can be represented by a change of an electric signal, and MXene has excellent conductivity and lower dielectric constant, which is more beneficial to the construction of an electrochemical sensor.
As one embodiment, the two-dimensional material MXene is Ti 3 C 2 、Ti 2 C、Nb 2 C、V 2 C、(Ti 0.5 Nb 0.5 ) 2 C、(V 0.5 Cr 0.5 ) 3 C 2 、Ti 3 CN or Ta 4 C 3 Any one of the following.
MXene has a highly porous structure and a high specific surface area, which enables it to provide more adsorption sites, contributing to adsorption and immobilization of DNA; the high specific surface area and porous structure of MXene contribute to the improvement of the adsorption capacity of DNA and the loading amount of the carrier, thereby improving the detection sensitivity. The MXene material has good biocompatibility in organisms, which is one of the advantages as a DNA carrier. The detection environment needs to be detected in a biological environment, and thus the biocompatibility of the carrier is very important for ensuring the safety of the biological environment. MXene as a carrier has low toxicity to cells and biological tissues, and helps to maintain the integrity of materials and the safety of biological environments.
As one embodiment, the mass ratio of MXene to calf thymus DNA is (100-1000): 1. the mass ratio of the MXene to calf thymus DNA is 100:1,200: 1,300: 1,400: 1,500: 1,600: 1,700: 1,800: 1,900: 1, 1000:1, or (100-1000): 1. The electric signal can be well expressed within the mass ratio range, and the electric signal has high specificity, stability and sensitivity.
A preparation method of a composite material based on MXene-DNA comprises the following steps:
and (3) dissolving and mixing calf thymus DNA and MXene, and then carrying out ultrasonic treatment to obtain the MXene-DNA composite material.
The hydrogen bond and the hydrophobic bond on the surface of the MXene can interact with the reverse adenine and cytosine base of calf thymus DNA to form the hydrogen bond and the hydrophobic bond, so that the DNA is firmly fixed on the surface of the MXene without special reaction, and the bonding of the hydrogen bond and the hydrophobic bond can be realized after simple mixing.
As one embodiment, the temperature during the ultrasonic treatment is 10-20 ℃, and the ultrasonic treatment time is 10-30min.
As one embodiment, MXene may be prepared by the method of: adding concentrated hydrochloric acid into lithium fluoride, uniformly stirring, adding an MXene base material, and continuing stirring reaction to obtain a multi-layer MXene after the reaction is finished; and carrying out ultrasonic vibration on the multi-layer MXene for 1-5 hours to obtain the two-dimensional material MXene.
Through the preparation method, the stacked MXene is dispersed, so that the subsequent probe preparation is more uniformly dispersed.
As one embodiment, the stirring reaction is carried out at 30-50℃for 6-48h.
As one embodiment, the method further comprises a water washing and alcohol washing step after the reaction. The clean multi-layer MXene material is obtained by water washing and alcohol washing.
As one embodiment, calf thymus DNA is heat treated prior to dissolution mixing with MXene, and calf thymus DNA is dissolved in water.
As one embodiment, the temperature of the heat treatment is 80-95 ℃. Preferably, the temperature of the heat treatment is 90-95 ℃. Further preferably, the temperature of the heat treatment is 95 ℃.
As one embodiment, the method further comprises a centrifugation step after the ultrasonic treatment. Centrifugation can leave unreacted calf thymus DNA in the supernatant, and the resulting lower precipitate is the MXene-DNA composite.
A sensor comprising an MXene-DNA based composite material of any one of the above.
The combination of calf thymus DNA and doxorubicin can change an electric signal due to the change of the DNA configuration, so that the detection can be performed by the change of the electric signal, the doxorubicin can be accurately detected in real time, and the detection result has a relatively obvious linear relationship. MXene has excellent conductivity and lower dielectric constant, and is more beneficial to the construction of an electrochemical sensor. The sensor has higher specificity, stability and sensitivity.
As one embodiment thereof, the MXene-DNA based composite material is coated on a glassy carbon electrode. The detection of doxorubicin knots by the MXene-DNA based composite material is shown in the form of an electrical signal, so that the detection can be directly carried out by coating the detection on a glassy carbon electrode and directly carrying out electrochemical detection by using the glassy carbon electrode.
A method of manufacturing a sensor comprising the steps of:
and (3) dripping the composite material based on the MXene-DNA on a glassy carbon electrode, and fixing by adopting a Nafion solution after air drying.
Specifically, after natural air drying, the dropwise added Nafion solution is fixed. Since simple MXene-loaded DNA is easily detached after being dried on an electrode, the Nafion is used for fixation modification, and the method has an effect without modification, but during the detection of an aqueous solution, the material is easily detached or detached entirely due to stirring, and the method is unstable.
Furthermore, nafion is an ion exchange polymer with highly fluorinated polytetrafluoroethylene groups that form a gel-like structure in aqueous solution and has a high negative charge density that enables Nafion to interact with either cationic or positive groups and provides a stable environment to immobilize and protect modified probes. And the DNA has the following effects on detection of small molecule drugs:
stability is enhanced: the Nafion modification can improve the stability of the probe, so that the probe can better withstand different experimental conditions, such as high temperature, high salt concentration or acid-base environment. Nafion can prevent the degradation or damage of the probe and prolong the service life of the probe.
Sensitivity is improved: nafion modification can enhance the interaction between the probe and the target molecule, thereby improving the detection sensitivity. The negative charge of Nafion can attract and localize cations or positive groups so that the cations or positive groups are closer to the interaction sites between the probes and the target molecules, and the progress of the binding reaction is promoted.
Inhibition of non-specific binding: the Nafion modification can reduce nonspecific binding and interference of background signals. The structure of Nafion can prevent non-specific binding substances from entering, thereby improving the selectivity and the specificity of the probe.
Promote immobilization of probes: nafion has gel-like properties and can firmly fix probes on the modified surface. This helps control the positioning and alignment of the probes, improving the reproducibility and consistency of the experiment.
As one embodiment, the concentration of the Nafion solution is 0.5% -5%. Preferably, the concentration of the Nafion solution is 0.5% -2%; further preferably, the concentration of Nafion solution is 0.5%.
As one embodiment, the glassy carbon electrode used was 3mm in diameter.
As one embodiment, the MXene-DNA composite material on the glassy carbon electrode is coated in an amount of 2-5 mug/mm 2 。
The application of the MXene-DNA based composite material or the sensor in the detection of the doxorubicin is realized by an electrochemical method.
Because of the relatively strong interaction between calf thymus DNA and doxorubicin, doxorubicin interferes with the structure and function of DNA by double-strand binding with DNA, and doxorubicin molecule binding with DNA has very strong specificity, while other small organic molecules have no interaction or weak interaction with DNA, the sensor prepared by the probe has specificity when detecting doxorubicin, thus the concentration change of doxorubicin can be accurately detected in complex body fluid environment. The novel two-dimensional material MXene with high conductivity and large surface area is used as a carrier to load calf thymus DNA with higher stability and rich sequencing data, and is used as a sensing signal probe to detect doxorubicin in real time and display the doxorubicin in the form of an electric signal, so that the doxorubicin is convenient to observe.
As one embodiment, the electrochemical method is a three-electrode system, the working electrode is the sensor, the counter electrode is a platinum wire electrode, and the reference electrode is Ag/AgCl.
Specific examples are described below.
Example 1
Thin layer MXene (Ti) 3 C 2 T x ) Is prepared from
1.5g of lithium fluoride powder is weighed, 20mL of 12M concentrated hydrochloric acid is added, and after uniform stirring, 1g of MXene base material Ti is added 3 AlC 2 Stirring for 24h at 40 ℃ to obtain clean multi-layer Ti by water washing and alcohol washing 3 C 2 T x A material; multilayer Ti 3 C 2 T x The material is vigorously vibrated at low temperature and is sonicated for 3 hours to obtain a few-layer Ti 3 C 2 T x Two-dimensional nanomaterial MXene.
Example 2
MXene-DNA composite material probe and preparation thereof
Firstly, thermally treating and dissolving calf thymus DNA, and mixing the treated DNA solution with the MXene solution of the example 1 in an oscillating way, wherein the mass ratio of the DNA to the MXene is 1:500; then, the ice bath is subjected to ultrasonic treatment at 16 ℃ for 20min, and then, the centrifugation is carried out to remove the supernatant, wherein the sediment is the MXene-DNA composite probe.
Example 3
MXene-DNA composite material probe and preparation thereof
Firstly, thermally treating and dissolving calf thymus DNA, and mixing the treated DNA solution with the MXene solution of the example 1 in an oscillating way, wherein the mass ratio of the DNA to the MXene is 1:100; then, the ice bath is subjected to ultrasonic treatment at 10 ℃ for 30min, and then, the ice bath is subjected to centrifugation, so that the supernatant is removed, and the sediment is the MXene-DNA composite probe.
Example 4
MXene-DNA composite material probe and preparation thereof
Firstly, thermally treating and dissolving calf thymus DNA, and mixing the treated DNA solution with the MXene solution of the example 1 in an oscillating way, wherein the mass ratio of the DNA to the MXene is 1:1000; then, the ice bath is subjected to ultrasonic treatment at 20 ℃ for 10min, and then, the ice bath is subjected to centrifugation, so that the supernatant is removed, and the sediment is the MXene-DNA composite probe.
Example 5
Preparation of MXene-DNA composite material sensor
Polishing a Glassy Carbon Electrode (GCE), and carrying out cyclic voltammetry test on the polished Glassy Carbon Electrode (GCE) in a 0.1M potassium ferricyanide/potassium chloride solution to judge whether the conditions are met; then the MXene-DNA composite probe of example 2 was prepared into a 5mg/ml solution, 4. Mu.L was dropped on a Glassy Carbon Electrode (GCE), and after it was naturally air-dried, 2. Mu.L of Nafion solution with a concentration of 0.5% was dropped on the glassy carbon electrode, and after it was air-dried, the sensor was obtained.
Comparative example 1
Polishing a Glassy Carbon Electrode (GCE), and carrying out cyclic voltammetry test on the polished Glassy Carbon Electrode (GCE) in a 0.1M potassium ferricyanide/potassium chloride solution to judge whether the conditions are met; then, the MXene prepared in example 1 was prepared into a 5mg/ml solution, 4. Mu.L was dropped on a Glassy Carbon Electrode (GCE) and naturally dried, then 2. Mu.L of Nafion solution with a concentration of 0.5% was dropped on the glassy carbon electrode and dried to obtain a sensor.
Comparative example 2
Polishing a Glassy Carbon Electrode (GCE), and carrying out cyclic voltammetry test on the polished Glassy Carbon Electrode (GCE) in a 0.1M potassium ferricyanide/potassium chloride solution to judge whether the conditions are met; then, the calf thymus DNA is dissolved by heat treatment, the treated DNA solution is prepared into 5mg/ml solution, 4 mu L of the solution is dripped on a Glassy Carbon Electrode (GCE), natural air drying is carried out, then 2 mu L of Nafion solution with the concentration of 0.5% is dripped on the glassy carbon electrode, and the sensor is obtained after the solution is air-dried.
Electrochemical detection of different concentrations of doxorubicin using the sensor of example 5
Pre-preparing doxorubicin mother solution (5 mg/ml), diluting 100 times for standby;
a three-electrode system was used, the working electrode was the sensor of example 5, the counter electrode was a platinum wire electrode, the reference electrode was Ag/Agcl (saturated KCL solution), a 100ml electrolytic cell was used, and 50ml deionized water (ph=7) was poured as the electrolyte;
adopting an i-t mode in an electrochemical workstation to detect, adding the prepared doxorubicin solution in the test process, and slowly stirring to ensure that the doxorubicin solution added in the test can be uniformly and fully received by a sensor to be detected in real time; a schematic diagram for real-time detection of doxorubicin is shown in FIG. 1;
in the test process, the prepared sensor is firstly linked with an electrode of an electrochemical workstation, and the mode of the electrochemical workstation is set as i-t. Then, the test is started, the test environment is an aqueous solution, 1ml of pre-prepared doxorubicin aqueous solution is dripped after the current value is stable, and 1ml of doxorubicin aqueous solution is continuously dripped after the current value is stable for about 60 seconds, and the current reduction phenomenon of the sensor of the embodiment 5 along with the increase of the doxorubicin concentration is obtained by 8 times of circulation, as shown in fig. 2.
As can be seen from the above test procedure and the results of fig. 2, the sensor of example 5 achieves that a change in concentration is detected in real time during the dripping. And the linear curve of the relation between the doxorubicin concentration and the current is obtained after treatment, as shown in figure 3, the current change of the sensor is in a linear relation within the range of 0.34nM-1.72nM of doxorubicin concentration, the linear equation is i= 29.28901-2.18572 (DOX), and the correlation coefficient R is shown in figure 3 2 = 0.99915. Indicating that the doxorubicin concentration in the liquid can be quantitatively measured from the electrical signal.
Electrochemical detection of different small molecules using the sensor of example 5
In order to study the specificity of the composite sensor in detecting doxorubicin, a three-electrode system was used, and the preconditions were the same as described above. In the test process, the prepared sensor is firstly linked with an electrode of an electrochemical workstation, and the mode of the electrochemical workstation is set as i-t. The test was then started, at which point the test environment was an aqueous solution. After the current value is stable, 1ml of a pre-prepared rhodamine water solution is added dropwise, after the current value is stable, 1ml of a rhodamine water solution is continuously added dropwise, and 1ml of an doxorubicin water solution is added dropwise after the current is stable, wherein each small molecule water solution is added dropwise for 2 times in a circulating way, namely rhodamine, doxorubicin, erythromycin, tetracycline hydrochloride, penicillin sodium and glutathione respectively; the concentration of all the small molecule water solutions is 0.05mg/ml; the i-t pattern of the sensor for detection of various small molecules is shown in FIG. 4.
And processing the data to prepare the variation of the current of different small molecules under the detection of the prepared composite material sensor shown in figure 5, wherein the variation is the largest when detecting the doxorubicin. The sensor of example 5 is shown to have specificity in binding to doxorubicin and relatively strong interaction with doxorubicin, and thus shows a large variation in electrical signal. Therefore, the sensor has specificity in detecting the doxorubicin, so that the concentration change of the doxorubicin can be accurately detected in a complex body fluid environment.
Adriamycin was detected by using the sensor of example 5, the sensor prepared by MXene alone of comparative example 1, and the sensor prepared by DNA alone of example 2 for comparison
Pre-preparing doxorubicin mother solution (5 mg/ml), diluting 100 times for standby;
a three-electrode system was used, the working electrode was the sensor of comparative example 1 or the sensor of comparative example 2, the counter electrode was a platinum wire electrode, the reference electrode was Ag/Agcl (saturated KCL solution), a 100ml electrolytic cell was used, and 50ml deionized water (ph=7) was poured as an electrolyte;
adopting an i-t mode in an electrochemical workstation to detect, adding the prepared doxorubicin solution in the test process, and slowly stirring to ensure that the doxorubicin solution added in the test can be uniformly and fully received by a sensor to be detected in real time;
in the test process, the prepared sensor is firstly linked with an electrode of an electrochemical workstation, and the mode of the electrochemical workstation is set as i-t. Then, the test is started, the test environment is an aqueous solution, 1ml of pre-prepared doxorubicin aqueous solution is dripped after the current value is stable, and 1ml of doxorubicin aqueous solution is continuously dripped after the current value is stable for about 60 seconds, and the circulation is performed for 8 times, and the result is shown in fig. 6 and 7.
As can be seen from fig. 6, when the pure MXene of comparative example 1 was supported on the glassy carbon electrode, there was no significant stepwise change in the current when the aqueous doxorubicin solution at the same concentration was detected, so that it could be judged that the DNA in the MXene-DNA composite of example 2 coated on the sensor of example 5 interacted with doxorubicin to cause a change in the electrical signal, not interference due to other factors.
As can be seen from fig. 7, the sensor of comparative example 2 has a weak current signal and a small current value compared to the sensor of example 5; this is because the sensor of example 5 is enhanced in electrical signal due to the addition of MXene and facilitates immobilization of DNA for realizing the device. In conclusion, the novel two-dimensional material MXene with high conductivity and large surface area is used as a carrier to load calf thymus DNA with higher stability and rich sequencing data as a sensing signal probe to detect the doxorubicin in real time, and the doxorubicin is expressed in the form of an electric signal, so that the doxorubicin is convenient to observe. The doxorubicin molecule is combined with DNA with strong specificity, and other small organic molecules have no interaction or weak interaction with DNA, so that the sensor prepared by the probe has specificity when detecting doxorubicin, thereby accurately detecting the concentration change of doxorubicin in complex body fluid environment. The sensor has higher sensitivity in the detection process, can detect the change of the electric signal in a shorter time, is favorable for realizing real-time detection in practical application, realizes the effect of real-time drug delivery, and is the optimal absorption state of the drug.
The above embodiments are only preferred embodiments of the present invention, and the scope of the present invention is not limited thereto, but any insubstantial changes and substitutions made by those skilled in the art on the basis of the present invention are intended to be within the scope of the present invention as claimed.
Claims (10)
1. The composite material based on the MXene-DNA is characterized by comprising a two-dimensional material MXene and calf thymus DNA, wherein the calf thymus DNA is connected to the surface of the two-dimensional material MXene through electrostatic action and/or hydrogen bonding action.
2. The MXene-DNA based composite material according to claim 1, wherein the two-dimensional material MXene is Ti 3 C 2 、Ti 2 C、Nb 2 C、V 2 C、(Ti 0.5 Nb 0.5 ) 2 C、(V 0.5 Cr 0.5 ) 3 C 2 、Ti 3 CN or Ta 4 C 3 Any one of the following.
3. A composite material based on MXene-DNA according to claim 1, wherein,
the mass ratio of the MXene to calf thymus DNA is (100-1000): 1.
4. the preparation method of the MXene-DNA based composite material is characterized by comprising the following steps of:
and (3) dissolving and mixing calf thymus DNA and MXene, and then carrying out ultrasonic treatment to obtain the MXene-DNA composite material probe.
5. The method for preparing a MXene-DNA based composite according to claim 4, wherein the temperature is 10-20deg.C and the time of the ultrasound is 10-30min.
6. A sensor comprising an MXene-DNA based composite material according to any one of claims 1-3.
7. The sensor of claim 6, wherein the MXene-DNA based composite is coated on a glassy carbon electrode.
8. A method of manufacturing a sensor, comprising the steps of:
and (3) dripping the composite material based on the MXene-DNA on a glassy carbon electrode, and fixing by adopting a Nafion solution after air drying.
9. Use of an MXene-DNA based composite material according to any one of claims 1-3 or a sensor according to claim 7 or claim 8 for the detection of doxorubicin, characterized in that doxorubicin is detected by electrochemical means.
10. Use according to claim 9, wherein the electrochemical method is a three-electrode system, the working electrode is the sensor according to claim 6 or claim 7, the counter electrode is a platinum wire electrode, and the reference electrode is Ag/AgCl.
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