CN114295587A - SPR sensor based on two-dimensional metal organic framework and preparation and application thereof - Google Patents
SPR sensor based on two-dimensional metal organic framework and preparation and application thereof Download PDFInfo
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Abstract
The invention discloses a SPR sensor based on a two-dimensional metal organic framework, wherein a Cu-TCPP 2D MOF sensitization layer is arranged on a gold chip, and the sensitization layer is prepared by the following steps: dissolving and adding copper nitrate, trifluoroacetic acid and polyvinylpyrrolidone into a first mixed solution of N, N-dimethylformamide and ethanol to form a second mixed solution; meanwhile, TCPP is dissolved in the first mixed solution, and then is dropwise added into the second mixed solution under stirring to form a third mixed solution; carrying out ultrasonic treatment on the third mixed solution and heating; cooling to room temperature, centrifuging the obtained Cu-TCPP 2D MOF solution, washing with ethanol, and dissolving in ethanol solution again for storage; dropping the stored Cu-TCPP 2D MOF solution on the surface of the gold chip for incubation, and washing the gold chip by deionized water to wash away the redundant Cu-TCPP 2D MOF solution; and then drying the gold chip by using nitrogen, and fixing the Cu-TCPP 2D MOF on the surface of the gold chip to form a Cu-TCPP 2D MOF sensitization layer.
Description
Technical Field
The invention relates to an SPR sensor, in particular to an SPR sensor based on a two-dimensional metal organic framework and preparation and application thereof.
Background
Surface Plasmon Resonance (SPR) spectroscopy is an optical technique for detecting the interaction of molecules on the surface of a gold film by monitoring the change in Refractive Index (RI). Compared with traditional analysis methods such as colorimetric method, fluorescence method and electrochemical analysis method, the non-labeling, simple and real-time characteristics of SPR enable the SPR to be widely applied to disease diagnosis, environmental and food safety supervision. The SPR signal response is outwards exponentially attenuated in the direction vertical to the interface, and is not sensitive to RI change exceeding 200nm of the sensing interface surface, so that the conventional SPR biosensor is difficult to detect macromolecular targets such as cells, bacteria and even exosomes. The detection based on the traditional sandwich method is complex in construction and needs multiple times of complex operations. Therefore, it is of great interest to construct SPR sensors based on a direct method without pre-labeling and outer layer sandwich.
To meet the demand for ultra-sensitive detection of trace disease biomarkers, nanomaterials have been proposed to modify the plasma sensing interface, thereby improving the sensitivity of SPR. By flowing functionalized AuNPs (gold nanoparticles) to the surface of the sensor chip to detect human igg, 25-fold signal enhancement was achieved, with excellent local surface plasmon resonance forming a strong coupling effect with the surface plasmon waves of the gold membrane. However, the disordered distribution of nanoparticles can interfere with the optical response of surface plasmon resonance, and the bulkiness and complexity of periodic nanoparticle structures limits their practical applications.
The two-dimensional material (graphene, molybdenum disulfide and the like) has larger specific surface area, high electron mobility and light absorption rate, and sensitive layer biomolecules for enhancing SPR signals can be constructed by the two-dimensional material under the condition of not interfering interaction, so that the sensitivity of SPR can be improved, and accurate detection is provided for trace biomarkers. Although the sensitivity of SPR sensors based on two-dimensional materials is improved, the detection accuracy and quality factor are reduced. Tunable and low-loss plasmons of graphene have attracted strong interest, but graphene suffers from the following disadvantages that will limit its direct application in SPR sensing: first, the relatively inefficient coupling between incident light and the graphene plasma is detrimental to sensitive measurements of the environment outside the sensing interface. Second, graphene may generate greater damping in Surface Plasmons (SPs) due to its high virtual dielectric constant, resulting in reduced detection accuracy. Third, one of the challenges of using graphene on SPR sensors is that it is prone to uncontrollable aggregation and difficult to homogenize and disperse well due to the high van der waals attraction between graphene sheets (5.9kJ mol/carbon), and thus the modification process of such two-dimensional materials in the sensing interface is more complicated. In conclusion, graphene alone is not the best candidate for high-sensitivity detection, and its application in SPR is mainly focused on graphene-other material composite structures. For the sensitization work of the current SPR sensor, an ideal two-dimensional material with a highly ordered internal structure, good photoelectric property, excellent solubility, dispersibility and easy modification is urgently needed to be found for constructing a huge potential sensor with the SPR performance.
Accordingly, those skilled in the art have endeavored to develop a SPR sensor having higher sensitivity.
Disclosure of Invention
In view of the above-mentioned drawbacks of the prior art, the present invention is directed to developing an SPR sensor with higher sensitivity.
In order to achieve the purpose, the invention discloses an ultrathin M-TCP 2D MOF (metalloporphyrin two-dimensional metal organic framework) nanosheet, and particularly a Cu-TCPP 2D MOF nanosheet which has an ultrathin thickness, a highly ordered structure, a plurality of active sites on the surface, a larger specific surface area and good solubility and dispersibility. The pi-stacked electroactive porphyrin molecules of TCPP-based 2D MOFs can perform charge transport in the MOF structure and have good electron mobility. Transient photocurrent generated by the 2D MOF under visible light irradiation promotes generation of efficient photo-generated carriers, and compared with graphene, the coupling effect between incident light and 2D MOF plasma is more excellent. In addition, Cu-TCPP has good in vitro and in vivo biocompatibility due to Cu2+The dd band transition and its two-dimensional ultra-thin property of (2) can produce excellent near infrared light absorption, and has many advantages in SPR sensitization.
The invention firstly provides a SPR sensor based on a two-dimensional metal organic framework, wherein a Cu-TCPP 2D MOF sensitization layer is arranged on a gold chip, and the sensitization layer is prepared by the following steps:
dissolving and adding copper nitrate, trifluoroacetic acid and polyvinylpyrrolidone into a first mixed solution of N, N-dimethylformamide and ethanol to form a second mixed solution; meanwhile, TCPP is dissolved in the first mixed solution, and then is dropwise added into the second mixed solution under stirring to form a third mixed solution; carrying out ultrasonic treatment on the third mixed solution and heating; cooling to room temperature, centrifuging the obtained Cu-TCPP 2D MOF solution, washing with ethanol, and dissolving in ethanol solution again for storage;
dropping the stored Cu-TCPP 2D MOF solution on the surface of the gold chip for incubation, and washing the gold chip by deionized water to wash away the redundant Cu-TCPP 2D MOF solution; and then drying the gold chip by using nitrogen, and fixing the Cu-TCPP 2D MOF on the surface of the gold chip to form a Cu-TCPP 2D MOF sensitization layer.
Further, in the first mixed solution, the volume ratio of the N, N-dimethylformamide to the ethanol is 3: 1; sonicating the third mixed solution for 10 minutes and heating to 80 ℃ for 3 hours; the incubation time on the gold chip surface is 30 minutes; the centrifugal treatment rotating speed is 8000r.p.m. and the time is 10 minutes.
Further, an incubation box is provided, the incubation box comprises a first box body and a second box body which can be buckled, a gold chip is placed in the first box body, and a Cu-TCPP 2D MOF solution is dripped into the second box body; then the first box body and the second box body are buckled, so that the gold film of the gold chip is soaked in the Cu-TCPP 2D MOF solution, the incubation box is clamped from two ends by rotating the clamping jaws, and the incubation box is rotated around the axis of the incubation box to carry out centrifugal incubation.
The invention also provides a preparation method of the SPR sensor based on the two-dimensional metal organic framework, which comprises the following steps:
dissolving and adding copper nitrate, trifluoroacetic acid and polyvinylpyrrolidone into a first mixed solution of N, N-dimethylformamide and ethanol to form a second mixed solution; meanwhile, TCPP is dissolved in the first mixed solution, and then is dropwise added into the second mixed solution under stirring to form a third mixed solution; carrying out ultrasonic treatment on the third mixed solution and heating; cooling to room temperature, centrifuging the obtained Cu-TCPP 2D MOF solution, washing with ethanol, and dissolving in ethanol solution again for storage;
dropping the stored Cu-TCPP 2D MOF solution on the surface of the gold chip for incubation, and washing the gold chip by deionized water to wash away the redundant Cu-TCPP 2D MOF solution; and then drying the gold chip by using nitrogen, and fixing the Cu-TCPP 2D MOF on the surface of the gold chip to form a Cu-TCPP 2D MOF sensitization layer.
Further, in the first mixed solution, the volume ratio of the N, N-dimethylformamide to the ethanol is 3: 1; sonicating the third mixed solution for 10 minutes and heating to 80 ℃ for 3 hours; the incubation time on the gold chip surface is 30 minutes; the centrifugal treatment rotating speed is 8000r.p.m. and the time is 10 minutes.
Further, an incubation box is provided, the incubation box comprises a first box body and a second box body which can be buckled, a gold chip is placed in the first box body, and a Cu-TCPP 2D MOF solution is dripped into the second box body; then the first box body and the second box body are buckled, so that the gold film of the gold chip is soaked in the Cu-TCPP 2D MOF solution, the incubation box is clamped from two ends by rotating the clamping jaws, and the incubation box is rotated around the axis of the incubation box to carry out centrifugal incubation.
The invention also provides an application of the SPR sensor based on the two-dimensional metal organic framework in detection of PD-L1 exosomes, which is characterized by comprising the following steps:
dissolving and adding copper nitrate, trifluoroacetic acid and polyvinylpyrrolidone into a first mixed solution of N, N-dimethylformamide and ethanol to form a second mixed solution; meanwhile, TCPP is dissolved in the first mixed solution, and then is dropwise added into the second mixed solution under stirring to form a third mixed solution; carrying out ultrasonic treatment on the third mixed solution and heating; cooling to room temperature, centrifuging the obtained Cu-TCPP 2D MOF solution, washing with ethanol, and dissolving in ethanol solution again for storage;
dropping the stored Cu-TCPP 2D MOF solution on the surface of the gold chip for incubation, and washing the gold chip by deionized water to wash away the redundant Cu-TCPP 2D MOF solution; then drying the gold chip by using nitrogen, and fixing the Cu-TCPP 2D MOF on the surface of the gold chip to form a Cu-TCPP 2D MOF sensitization layer;
loading a gold chip in an SPR instrument, then flowing the peptide with the function of specifically capturing PD-L1 exosomes on the surface of the gold chip with a sensitization layer, and fixing the peptide on the surface of the gold chip through pi-pi stacking; the PD-L1 exosome test solution is then flowed over the gold chip surface and detected by SPR instruments.
Further, in the first mixed solution, the volume ratio of the N, N-dimethylformamide to the ethanol is 3: 1; sonicating the third mixed solution for 10 minutes and heating to 80 ℃ for 3 hours; the incubation time on the gold chip surface is 30 minutes; the centrifugal treatment rotating speed is 8000r.p.m. and the time is 10 minutes.
Further, the peptide was flowed over the surface of the gold chip at a flow rate of 5. mu.L/min; the PD-L1 exosome solution to be tested flows through the surface of the gold chip at the flow rate of 5 muL/min.
Further, an incubation box is provided, the incubation box comprises a first box body and a second box body which can be buckled, a gold chip is placed in the first box body, and a Cu-TCPP 2D MOF solution is dripped into the second box body; then the first box body and the second box body are buckled, so that the gold film of the gold chip is soaked in the Cu-TCPP 2D MOF solution, the incubation box is clamped from two ends by rotating the clamping jaws, and the incubation box is rotated around the axis of the incubation box to carry out centrifugal incubation.
The invention uses the Cu-TCPP 2D MOF which has the advantages of ultrathin thickness, highly ordered structure, many surface active sites, large specific surface area, excellent electron mobility and light absorption and the like as the sensitization layer, thereby obviously improving the SPR sensitivity. According to the invention, a direct SPR detection method based on the 2D MOF sensitization layer is constructed without constructing a complex traditional sandwich, and target analysis and detection can be carried out only by three-step sample injection. The multifunctional peptide used in the invention has high specificity with the target, and the detection sample can complete the analysis and detection without pretreatment.
The conception, the specific structure and the technical effects of the present invention will be further described with reference to the accompanying drawings to fully understand the objects, the features and the effects of the present invention.
Drawings
FIG. 1 is an atomic force microscope image of a step-wise build of a sensing interface in a preferred embodiment of the invention;
FIG. 2 is a calibration chart of sensor selectivity verification and SPR angle change caused by different concentrations of PD-L1 exosomes in a preferred embodiment of the present invention.
FIG. 3 is a schematic view of the operation of the incubation box in a preferred embodiment of the present invention.
Detailed Description
The technical contents of the preferred embodiments of the present invention will be more clearly and easily understood by referring to the drawings attached to the specification. The present invention may be embodied in many different forms of embodiments and the scope of the invention is not limited to the embodiments set forth herein.
The determination method and the steps for detecting PD-LI exosomes using SPR sensors based on Cu-TCPP 2D MOF in this example are as follows:
(1) synthesis of Cu-TCPP 2D MOF:
a mixed solution of 12mL N, N-Dimethylformamide (DMF) and ethanol (V: V ═ 3:1) was added to a 20mL bottle, and then copper nitrate (3.6mg, 0.015mmol), trifluoroacetic acid (1.0M × 10 μ L), polyvinylpyrrolidone (PVP, 10.0mg) were added and dissolved therein. Meanwhile, TCPP (4.0mg, 0.005mmol) was dissolved in a mixed solution of 4ml dmdff and ethanol (V: V ═ 3:1), and then added dropwise to the above mixed solution with stirring. Thereafter, the mixed solution was sonicated for 10 minutes and heated to 80 ℃ for 3 hours. After heating ceased, the solution was cooled to room temperature and the resulting magenta Cu-TCPP 2D MOF solution was centrifuged at 8000r.p.m. for 10 minutes and washed twice with ethanol. Finally the obtained Cu-TCPP 2D MOF was re-dissolved in 10ml ethanol solution and stored at 4 ℃.
(2) SPR sensor performance analysis
Different refractive index measurements were analyzed, including deionized water and 1-16.7 wt% sodium chloride solution (RI 1.33330, 1.33500, 1.33675, 1.34170, 1.34986, 1.36370). 300 μ different mass fractions of sodium chloride solution were flowed to the gold chip and rinsed with deionized water. The SPR angular displacement is measured and collected throughout the monitoring process. SPR signal changes were compared between bare gold chips and two-dimensional MOF-modified gold chips using the same concentration of multifunctional peptide and exosomes.
(3) Construction of SPR sensor chip for detecting PD-L1 exosome
50 μ L of a 2DMOF nanosheet solution at an appropriate concentration was dropped onto the surface of the treated gold chip. After 30 minutes of incubation, the gold chips were washed with deionized water to wash away excess 2DMOF nanosheet solution. And drying the gold chip by using nitrogen again, and fixing the two-dimensional MOF nanosheet on the surface of the gold chip. The modified gold chip was fixed in the SPR instrument. Then 300. mu.L of 100. mu.g/mL peptide having an anti-contamination function and specifically capturing the PD-L1 exosome function was flowed through the surface of 2D MOF nanoplate at a flow rate of 5. mu.L/min and the peptide was immobilized on the surface of the gold chip by π - π stacking. Then 300. mu.L of PD-L1 exosomes at different concentrations, which specifically bind to the peptide, were flowed over the gold chip surface at a flow rate of 5. mu.L/min. The buffer solution used for baseline was deionized water and the entire sampling process was performed under the monitoring of the SPR instrument.
SPR monitoring of PD-L1 exosomes:
the PD-L1 exosome concentrations used were: 104、5×104、105、5×105、106、5×106exosomes/mL.
And (3) testing conditions are as follows: SPR measurements of different concentrations of PD-L1 exosomes were performed using an SPR meter at room temperature.
Referring to the drawings, FIG. 1 is an atomic force microscope image of a stepwise build of a sensing interface. The 2D images and cross-sectional thickness maps of gold chips (a), 2D MOF-modified chips (B), exosome-capturing sensory interfaces (C) were characterized by atomic force microscopy, respectively. Two-dimensional MOFs were modified on flat smooth bare gold sheets and the layered structure of Cu-TCPP was again confirmed by atomic force microscopy. The morphological structure of the bare gold chip showed good uniformity with a roughness of 5 nm. As is clear from FIG. 1B, the MOF is a two-dimensional sheet-like structure with a relatively uniform morphology and a thickness of about 10nm (minus the bare gold sheet roughness of 5 nm). Then, the sensing interface for capturing the exosomes is characterized, and the exosomes with the length of 120nm can be clearly observed to exist on the 2D MOF, which indicates that the construction of the sensing interface is successful.
FIG. 2 is a calibration chart of sensor selectivity verification and SPR angle change caused by different concentrations of PD-L1 exosomes. It is well known that a variety of interfering substances are present in the blood environment,such as impure proteins, ascorbic acid, glutathione, glucose, etc. The recognition domain (FHYQRDTPKSYN) in the peptide was shown to have a strong binding capacity to PD-L1 with a dissociation constant (KD) of 0.51. mu.M. Whether the peptide recognition domain can specifically capture the high-expression PD-L1 extracellular domain on an exosome is one of the key parts for constructing an accurate and sensitive blood environment exosome sensor. FIG. 2A, B records signal changes caused by interfering substances in blood (GSH, AA, Glu, BSA) and exosomes flowing across the SPR sensor chip surface. The mean values of the changes in SPR signal by the four interfering substances were only 7m DEG, -1m DEG, 8m DEG and 15m DEG, respectively, compared to the change in signal at 97m DEG caused by the target PD-L1 exosome, with a p-value of less than 0.01 indicating a very significant difference. It can be seen that the four interfering substances do not have a significant effect on the SPR sensor. These results confirm that the fabricated sensor has excellent specificity for exosomes. To verify the linear relationship between the amount of the substance detected and the signal response of the SPR sensor, the PD-L1 exosome concentrations were used as follows: 104、5×104、105、5×105、106、5×106exosomes/mL. FIG. 2C, D is an SPR curve of changes in signal caused by gradient concentration targets flowing across the surface of a sensor chip and a linear fit of the resulting data. The SPR angular displacement increases with increasing PD-L1 exosome concentration, and the change in SPR angle has a good linear relationship to the logarithm of PD-L1 exosome concentration. Linear equation is Δ θ ═ 0.0324lgCExo0.11984, the linear correlation coefficient is 0.989, the detection limit is 16.7 exosomes/mL, and the detection requirement is met.
As shown in figure 3, in order to fix the two-dimensional MOF nano-sheets on the surface of the gold film 3 of the gold chip 4 more quickly and tightly and save the incubation time, an incubation box is provided, and comprises a first box body 1 and a second box body 2 which can be buckled. And then a gold chip 4 is placed in the first box body 1, and a 2DMOF nanosheet solution 5 is dripped into the second box body 2. Then buckle first box body 1 and second box body 2 for the gold membrane 3 of gold chip 4 soaks in 2DMOF nanosheet solution, through rotatory clamping jaw 6 from both ends centre gripping incubation box, and it is rotatory around its axis and carry out centrifugal incubation, make two-dimentional MOF nanosheet in 2DMOF nanosheet solution deposit more fast more closely and fix on gold membrane 3 surface of gold chip 4.
The invention uses the Cu-TCPP 2D MOF with various advantages of ultrathin thickness, highly ordered structure, many surface active sites, large specific surface area, excellent electron mobility and light absorption and the like as the sensitization layer, thereby obviously improving the SPR sensitivity. The method constructs a direct SPR detection method based on the 2D MOF sensitization layer, the used multifunctional peptide has multiple functions and a whole body of anchoring the 2D MOF, preventing interface nonspecific adsorption, capturing PD-L1 exosomes specifically and the like, the complex and fussy interface construction can be omitted, and target analysis and detection can be carried out only by 3 steps. The method has high specificity, selectivity, simplicity and sensitivity, and can detect trace amount of PD-L1 exosomes. The constructed simple and rapid SPR sensor chip has good expandability and wide prospects, and the method provides a new platform for clinically detecting a few of micro target biomolecules.
The foregoing detailed description of the preferred embodiments of the invention has been presented. It should be understood that numerous modifications and variations could be devised by those skilled in the art in light of the present teachings without departing from the inventive concepts. Therefore, the technical solutions available to those skilled in the art through logic analysis, reasoning and limited experiments based on the prior art according to the concept of the present invention should be within the scope of protection defined by the claims.
Claims (10)
1. A SPR sensor based on a two-dimensional metal organic framework is characterized in that a Cu-TCPP 2D MOF sensitization layer is arranged on a gold chip, wherein the sensitization layer is prepared through the following steps:
dissolving and adding copper nitrate, trifluoroacetic acid and polyvinylpyrrolidone into a first mixed solution of N, N-dimethylformamide and ethanol to form a second mixed solution; meanwhile, TCPP is dissolved in the first mixed solution, and then is dropwise added into the second mixed solution under stirring to form a third mixed solution; carrying out ultrasonic treatment on the third mixed solution and heating; cooling to room temperature, centrifuging the obtained Cu-TCPP 2D MOF solution, washing with ethanol, and dissolving in ethanol solution again for storage;
dropping the stored Cu-TCPP 2D MOF solution on the surface of the gold chip for incubation, and washing the gold chip by deionized water to wash away the redundant Cu-TCPP 2D MOF solution; and then drying the gold chip by using nitrogen, and fixing the Cu-TCPP 2DMOF on the surface of the gold chip to form a Cu-TCPP 2D MOF sensitization layer.
2. The two-dimensional metal organic framework-based SPR sensor according to claim 1, wherein the volume ratio of N, N-dimethylformamide to ethanol in said first mixed solution is 3: 1; sonicating the third mixed solution for 10 minutes and heating to 80 ℃ for 3 hours; the incubation time on the gold chip surface is 30 minutes; the centrifugal treatment rotating speed is 8000r.p.m. and the time is 10 minutes.
3. The SPR sensor based on a two-dimensional metal organic framework according to claim 1, wherein an incubation box is provided, the incubation box comprises a first box body and a second box body which can be buckled, a gold chip is placed in the first box body, and a Cu-TCPP 2D MOF solution is dripped into the second box body; then the first box body and the second box body are buckled, so that the gold film of the gold chip is soaked in the Cu-TCPP 2D MOF solution, the incubation box is clamped from two ends by rotating the clamping jaws, and the incubation box is rotated around the axis of the incubation box to carry out centrifugal incubation.
4. A preparation method of an SPR sensor based on a two-dimensional metal organic framework is characterized by comprising the following steps:
dissolving and adding copper nitrate, trifluoroacetic acid and polyvinylpyrrolidone into a first mixed solution of N, N-dimethylformamide and ethanol to form a second mixed solution; meanwhile, TCPP is dissolved in the first mixed solution, and then is dropwise added into the second mixed solution under stirring to form a third mixed solution; carrying out ultrasonic treatment on the third mixed solution and heating; cooling to room temperature, centrifuging the obtained Cu-TCPP 2D MOF solution, washing with ethanol, and dissolving in ethanol solution again for storage;
dropping the stored Cu-TCPP 2D MOF solution on the surface of the gold chip for incubation, and washing the gold chip by deionized water to wash away the redundant Cu-TCPP 2D MOF solution; and then drying the gold chip by using nitrogen, and fixing the Cu-TCPP 2DMOF on the surface of the gold chip to form a Cu-TCPP 2D MOF sensitization layer.
5. The method for preparing a SPR sensor based on a two-dimensional metal organic framework according to claim 4, wherein in the first mixed solution, the volume ratio of N, N-dimethylformamide to ethanol is 3: 1; sonicating the third mixed solution for 10 minutes and heating to 80 ℃ for 3 hours; the incubation time on the gold chip surface is 30 minutes; the centrifugal treatment rotating speed is 8000r.p.m. and the time is 10 minutes.
6. The preparation method of the SPR sensor based on the two-dimensional metal organic framework, according to claim 4, wherein an incubation box is provided, the incubation box comprises a first box body and a second box body which can be buckled, a gold chip is placed in the first box body, and a Cu-TCPP 2D MOF solution is dripped into the second box body; then the first box body and the second box body are buckled, so that the gold film of the gold chip is soaked in the Cu-TCPP 2D MOF solution, the incubation box is clamped from two ends by rotating the clamping jaws, and the incubation box is rotated around the axis of the incubation box to carry out centrifugal incubation.
7. The application of an SPR sensor based on a two-dimensional metal-organic framework in detecting PD-L1 exosomes is characterized by comprising the following steps:
dissolving and adding copper nitrate, trifluoroacetic acid and polyvinylpyrrolidone into a first mixed solution of N, N-dimethylformamide and ethanol to form a second mixed solution; meanwhile, TCPP is dissolved in the first mixed solution, and then is dropwise added into the second mixed solution under stirring to form a third mixed solution; carrying out ultrasonic treatment on the third mixed solution and heating; cooling to room temperature, centrifuging the obtained Cu-TCPP 2D MOF solution, washing with ethanol, and dissolving in ethanol solution again for storage;
dropping the stored Cu-TCPP 2D MOF solution on the surface of the gold chip for incubation, and washing the gold chip by deionized water to wash away the redundant Cu-TCPP 2D MOF solution; then drying the gold chip by using nitrogen, and fixing the Cu-TCPP 2DMOF on the surface of the gold chip to form a Cu-TCPP 2D MOF sensitization layer;
loading a gold chip in an SPR instrument, then flowing the peptide with the function of specifically capturing PD-L1 exosomes on the surface of the gold chip with a sensitization layer, and fixing the peptide on the surface of the gold chip through pi-pi stacking; the PD-L1 exosome test solution is then flowed over the gold chip surface and detected by SPR instruments.
8. The use of the two-dimensional metal-organic framework based SPR sensor of claim 7 for the detection of PD-L1 exosomes, wherein, in the first mixed solution, the volume ratio of N, N-dimethylformamide to ethanol is 3: 1; sonicating the third mixed solution for 10 minutes and heating to 80 ℃ for 3 hours; the incubation time on the gold chip surface is 30 minutes; the centrifugal treatment rotating speed is 8000r.p.m. and the time is 10 minutes.
9. Use of a two-dimensional metal-organic framework based SPR sensor according to claim 7 for the detection of PD-L1 exosomes, wherein the peptide is flowed over the surface of the gold chip at a flow rate of 5 μ L/min; the PD-L1 exosome solution to be tested flows through the surface of the gold chip at the flow rate of 5 muL/min.
10. The application of the SPR sensor based on the two-dimensional metal-organic framework in detecting PD-L1 exosomes, according to claim 7, wherein an incubation box is provided, the incubation box comprises a first box body and a second box body which can be buckled, a gold chip is placed in the first box body, and a Cu-TCPP 2D MOF solution is dripped into the second box body; then the first box body and the second box body are buckled, so that the gold film of the gold chip is soaked in the Cu-TCPP 2D MOF solution, the incubation box is clamped from two ends by rotating the clamping jaws, and the incubation box is rotated around the axis of the incubation box to carry out centrifugal incubation.
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