CN109540991A - Functional metal organic framework material, FKN sensor of its building and preparation method thereof - Google Patents
Functional metal organic framework material, FKN sensor of its building and preparation method thereof Download PDFInfo
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Abstract
The present invention relates to a kind of functional metal organic framework materials, the FKN sensor of its building and preparation method thereof.The gold nanoparticle functional metal organic backbone for having synthesized a kind of sulfonated calixarenes cladding of novel porosity, constructs sensitive FKN electrochemical sensor, detects the lowest limit up to 7.4 pg/mL.Advantage is embodied in first: pSC-AuNPs multilayer film and shows good biocompatibility and high electron-transporting, and electrochemical signals obtain significant improvement, and pSC-AuNPs identifies target protein FKN in conjunction with FKN antibody.Second: on the surface MOFs, aptamer modified pSC-AuNPs can be fixed by Host-guest Recognition, improve the load factor of aptamer, using the excellent catalytic performance of MOF, obtain sensitive H2O2Restore electric signal.Be compared with other methods, this method have the characteristics that preparation is easy, equipment is simple, it is time-consuming less, response is fast and finished product stability is high.The present invention detects FKN using the method for MOFs material and electrochemistry for the first time, realizes the Sensitive Detection to the sample for simulating practical FKN.
Description
Technical field
The present invention relates to a kind of functional metal organic framework materials, the FKN sensor of its building and preparation method thereof.
Background technique
Fractalkine (FKN, also referred to as CX3CL1) has CX3C chemotactic factor (CF) structural domain, and thin in the endothelium of activation
It is expressed in born of the same parents, neuron, dendritic cells and vascular smooth muscle cells.FKN is a kind of transmembrane protein, by with specific receptor
CX3CR1 in conjunction between carrying out mediated cell adherency and signal transduction.It has recently been demonstrated that other than chemotaxis and adhesiveness,
FKN plays indispensable role in certain diseases such as Alzheimer disease, inflammation and obesity.FKN is detection Ba Leisi
The biomarker of special esophagus disease.So far, most of researchs about FKN concentrate on the machine to interact with various diseases
In system, and the method for quantitative detection FKN is seldom.FKN mainly passes through the reagent based on enzyme-linked immunosorbent assay (ELISA)
Box measures.In order to delicately and quantitatively measure FKN, it is still desirable to construct new system.
Metal-organic framework materials (MOFs) are a kind of coordination polymers quickly grown nearly ten years.It is usually with metal
Ion is tie point, and using organic ligand as linking group material, being formed has porous, high surface area, structure and function diversity
And the three-dimensional framework of remarkable thermal stability.Due to these excellent performances, MOFs has been used for gas storage, separates, catalysis,
Energy, the fields such as drug conveying and chemical sensitisation.Gold nano grain (AuNPs) is widely used in various biosensors, has aobvious
Optics, electricity and chemical property, high chemical stability are readily synthesized and become novel hybride nanometer with surface-functionalized
The important component of material.The MOFs of metal nanoparticle functionalization shows peroxidase sample activity.
Calixarenes typically refers to the big ring being made of the phenol units of methylene-bridged, and referred to as crown ether and cyclodextrin it
Third generation host compound afterwards.Up to the present, the mixing nano material of You great Huan supermolecule and AuNPs composition has obtained
Research well.Sulfonated calixarenes (pSC) is a kind of Water-Soluble Calixarenes with rigidity and open cavity, to guest molecule
With sizable binding ability, it is used as the stabilizer of metal nanoparticle.Therefore, the nanoparticle that calixarenes is modified is
As the hot topic in chemical sensor.
Therefore, we be badly in need of preparing it is a kind of cost is relatively low, it is highly selective, can the non-enzymatic of fast and reliable safety detection FKN pass
Sensor, for the detection of clinical treatment, the diagnosis of disease early period is of great significance for this.
Summary of the invention
One of the objects of the present invention is to provide a kind of functional metal organic framework materials.
The second object of the present invention is to provide the FKN sensor constructed by the framework material.
The third object of the present invention is the preparation method of the sensor.
In order to achieve the above objectives, the present invention uses following mechanism: having synthesized a kind of sulfonated calixarenes of novel porosity
The gold nanoparticle functional metal organic backbone (pSC-AuNPs/MOFs) of cladding, constructs sensitive FKN sensor, is used in combination
Make oxidation H2O2.On the one hand, different layers of conduction pSC-AuNPs films are fixed on gold electrode table by host-guest interaction
Face.PSC-AuNPs multilayer film shows good biocompatibility and high electron-transporting, by successively assembling pSC-AuNPs,
Electrochemical signals obtain significant improvement, and pSC-AuNPs identifies target protein FKN in conjunction with FKN antibody.On the other hand, in MOFs
Surface, aptamer modified pSC-AuNPs can be fixed by Host-guest Recognition.From Cu (II) to Cu, the electronics of (III) turns
Shifting can be by MOFs to H2O2The catalysis of reduction and improved, cause significant signal to read.PSC-AuNPs/MOFs is not only
With quick electron transport ability, and increase the load capacity that can identify the FKN aptamer of FKN.Design in this way,
The protrusion analysis ability of pSC-AuNPs/MOFs signal amplification strategy provides potential application for the clinical detection of FKN.
According to above-mentioned mechanism, the present invention adopts the following technical scheme:
A kind of functional metal organic framework material, it is characterised in that the structure of the framework material are as follows: with Cu2+Centered on gold
Belong to ion, 2- amino terephthalic acid (TPA) is that ligand has synthesized a kind of metal organic frame (MOFs) material under conditions of solvent heat
Expect Cu (BDC-NH2)2, BDC expression p-phthalic bridge；The copper base metal skeleton Surface-modification of Nanoparticles has amino group.
A kind of FKN sensor is constructed, it is characterised in that the sensing using above-mentioned functional metal organic framework material
Device are as follows: using the fixed FKN antibody of the pSC-AuNPs self-assembled multilayer film in gold electrode surfaces as biorecognition surface；Simultaneously
A kind of gold nanoparticle functionalization metal-organic framework materials of porosity sulfonic group calixarenes cladding are prepared for for signal post
Label are to be catalyzed H2O2。
A method of preparing above-mentioned FKN sensor, it is characterised in that the specific steps of this method are as follows:
It a. is 10 by 2.8mL concentration-2Mol/L pSC and concentration are 2.63 × 10-2Mol/L gold chloride (HAuCl4) be protected from light and stir
Mix 20min.Being rapidly added 2.8mL concentration later is 10-2Sodium borohydride (the NaBH of mol/L4), it is stirred to react to the color of solution
Claret is eventually become from the faint yellow dark brown that becomes, stops reaction, is protected from light standing after 5-8 hours the pSC- prepared
AuNPs be transferred in brown bottle be stored in 4 DEG C it is spare；
B. to prepare pSC-AuNPs/MOFs, then 100 μ L pSC-AuNPs are instilled into gained MOFs stirring 10-12 hours
By 200 μ L, 1 μM of NH2The FKN Apt of sealing end is added in above-mentioned pSC-AuNPs/MOFs solution, passes through stirring 10-12 hours
The Apt/pSC-AuNPs/MOFs of acquisition is modified to the three-dimensional electrode surface constructed in three-dimensional gold nano afterwards, utilizes MOFs peroxidating
The effect of hydrogen analogue enztme is catalyzed H2O2Decomposition achieve the purpose that detect FKN.
Above-mentioned metal organic frame the preparation method comprises the following steps: stirring under, by polyvinylpyrrolidone, 2- amino terephthaldehyde
Acid and Cu (NO3)2·3H2O is dissolved in n,N-Dimethylformamide and ethyl alcohol by the molar ratio of 1:3:10 and mixes by the volume ratio of 1:1
It closes in object, mixes well, heated 4-6 hours at 100 DEG C；Remove supernatant, then be added DMF to dissolve gained sediment,
By the solution at 100 DEG C of heating 8-10 hours to remove unreacted reagent, centrifuge separation obtains metal organic frame.
The present invention utilizes the FKN sensor of metal-organic framework materials building, is fixed on the pSC-AuNPs of gold electrode surfaces
Multilayer film shows good biocompatibility and high electron-transporting, and aptamer modified pSC-AuNPs can pass through Subjective and Objective
Identification is fixed on MOFs material, by MOFs to H2O2Catalysis and achieve the purpose that detect FKN.The MOFs of synthesis is tied
Structure has the catalytic activity of simulation catalase, the partial size of the MOFs nano particle: about 1.5um.pSC-AuNPs/
MOFs not only has quick electron transport ability, but also increases the load capacity that can identify the FKN aptamer of FKN.In electrification
Catalysis is carried out to hydrogen peroxide under the conditions of and has the function that FKN sensor.
The FKN of various concentration carries out Electrochemical Scanning in the hydrogen peroxide PBS buffer solution containing 10mM, acquisition
DPV map.It can see the increase with FKN concentration, the redox reaction peak position for corresponding to FKN at 0.01V voltage
Increasing.By the detection of the FKN solution to 10pg/mL-10 μ g/mL concentration, FKN concentration and redox peak current are established
Linear relationship, research finds that the sensor has good reproducibility, stability and selectivity, finally molten using the serum of mark-on
Liquid carries out the detection of actual sample, the variation of electrochemical signals can be also detected in serum, illustrates to utilize MOFs peroxidating
The sensor of hydrogen analogue enztme can detect FKN, and be applied in the detection of actual sample.
The advantages and features of the present invention is as described below: the present invention detects FKN using MOFs material for the first time, is also simultaneously
The method of electrochemistry detects FKN for the first time.The present invention is in order to develop sensitive FKN electrochemica biological sensor, benefit
The pSC-AuNPs self-assembled multilayer film in gold electrode surfaces is used to fix FKN antibody as biorecognition surface.It is prepared for one simultaneously
The gold nanoparticle functionalization metal-organic framework materials of the novel porosity sulfonic group calixarenes cladding of kind are used for signal label
To be catalyzed H2O2.The pSC-AuNPs multilayer film for being fixed on gold electrode surfaces shows good biocompatibility and high electron-transport
Property, aptamer modified pSC-AuNPs can be fixed on MOFs material by Host-guest Recognition, by MOFs to H2O2's
It is catalyzed and achievees the purpose that detect FKN.The MOFs structure of synthesis has the catalytic activity of simulation catalase, the MOFs
The partial size of nano particle: about 1.5um.PSC-AuNPs/MOFs not only has quick electron transport ability, but also increases
It can identify the load capacity of the FKN aptamer of FKN.Catalysis is carried out to hydrogen peroxide under electrochemical conditions and reaches FKN sensor
Effect.PSC-AuNPs multilayer film has good biocompatibility and high electron-transporting.In conjunction with the good catalysis energy of MOFs
Power realizes quantitative FKN with sensitive electrochemical immunosensor and detects, and detection is limited to 7.4pg/mL.
The present invention constructs a kind of novel analog enzyme sensor with metal-organic framework materials detection FKN, and metal is had
Machine framework material is applied in electrochemical interface detection, achieves satisfied result.This detection method can be observed intuitively
The numerical value of specific peak current detects the concentration of FKN, while effectively can also play inspection in complicated actual sample detection
Survey the effect of FKN.The analogue enztme FKN sensor that we construct provides more for the detection of FKN in disease surveillance and real life
For convenient and fast method, same principle is also extrapolated to the detection of other materials, and application prospect is extensive.
Detailed description of the invention
Fig. 1 is scanning of the metal-organic framework materials system modified electrode of the invention constructed in the FKN of various concentration
DPV figure.
Fig. 2 is the scanning electron microscope diagram of metal-organic framework materials.
Specific embodiment
After now specific embodiments of the present invention are described in.
Embodiment 1
The FKN sensor of functional metal organic framework material, its building in the present embodiment and preparation method thereof step
It is as follows:
(1) gold nano grain of pSC functionalization the preparation method comprises the following steps: by volume be 250mL round-bottomed flask load onto brandreth
The ultrapure water of 126mL and stirring is added in platform, and it is 10 that 2.8mL concentration, which is then added,-2Mol/L pSC and concentration are 2.63 × 10- 2Mol/L gold chloride (HAuCl4) it is protected from light stirring 20min.Being rapidly added 2.8mL concentration later is 10-2The sodium borohydride of mol/L
(NaBH4), stop after being vigorously stirred two hours, the color that solution can be observed in whole process becomes dark brown most from faint yellow
After become claret, the gold nano grain of the pSC functionalization prepared is transferred to brown bottle after standing 5 hours after stopping by stirring
In be stored in 4 DEG C it is spare.
(2) preparation method of metal-organic framework materials: firstly, under stiring dissolving 0.20g polyvinylpyrrolidone
In the mixture of 4mL n,N-Dimethylformamide and 4mL ethyl alcohol.Then, by the 2- amino terephthalic acid (TPA) of 5.43mg and
Cu (the NO of 24.20mg3)2·3H2O is mixed into the DMF of 4mL and is added in said mixture.By said mixture ultrasound
After twenty minutes, it is heated 5 hours at 100 DEG C in autoclave for processing.Then, remove supernatant, 20mL is then added
DMF is to dissolve gained sediment.Solution is further heated to 8 hours at 100 DEG C to remove unreacted reagent.Finally, logical
It crosses centrifugation and obtains resulting MOFs, and store at 4 DEG C for further use.
(3) 100 μ L pSC-AuNPs the preparation method of FKN sensor: are instilled into gained MOFs stirring 10-12 hours to make
Standby pSC-AuNPs/MOFs, then by 200 μ L, 1 μM of NH2The FKN aptamer of sealing end is added to above-mentioned pSC-AuNPs/MOFs solution
In, pass through the three-dimensional electricity for constructing the Apt/pSC-AuNPs/MOFs modification of acquisition in three-dimensional gold nano after stirring 10-12 hours
Pole surface is catalyzed H using the effect of MOFs mimichydrogen peroxidase2O2Decomposition achieve the purpose that detect FKN.
Entire sensor is made of the electrochemical workstation of three-electrode system, and wherein reference electrode is saturated calomel electrode,
It is platinum electrode to electrode, working electrode is the gold electrode for being modified with three-dimensional gold nano structure, and prepare liquid is used as electrolyte.
Electrochemical sensor analogue enztme detects FKN: being fixed using the pSC-AuNPs self-assembled multilayer film in gold electrode surfaces
FKN antibody is as biorecognition surface.It is prepared for a kind of Jenner's grain of rice of novel porosity sulfonic group calixarenes cladding simultaneously
Son functionalization metal-organic framework materials are catalyzed H for signal label2O2.Utilize the good biofacies of pSC-AuNPs multilayer film
Capacitive and high electron-transporting realize the purpose of Electrochemical Detection FKN in conjunction with the good catalytic ability of MOFs.
Electrolyte conditions: use the PBS buffer solution for containing 1mM hydrogen peroxide of pH=7.4 as the electricity of experiment detection
Solve liquid.
The concentration range of FKN: 10pg/mL-10 μ g/mL
Referring to attached drawing, Fig. 1 is metal-organic framework materials system modified electrode the sweeping in the FKN of various concentration of building
Retouch DPV figure.Fig. 2 is the scanning electron microscope diagram of metal-organic framework materials.As seen from the figure, with the increase of FKN concentration,
H at 0.01V2O2Oxidation-reduction potential peak value also increases with it, and may determine that by the presence or absence of peak current and numerical values recited to be measured
It whether there is the concentration of FKN and FKN in system.It is able to observe that the metal of synthesis is organic by scanning electron microscope
The good pattern of framework material, is easy to surface modification.
The present invention simulates biological enzyme by metal-organic framework materials and detects FKN, is compared with other methods, preparation method letter
Just, complicated equipment is not needed, the consuming time is few, and analogue enztme is simple to save compared to biological enzyme, low in cost, effect stability.This
One detection method has largely expanded the application of three-dimensional gold nano, it will the gold nano grain system for keeping building three-dimensional exists
It is more widely applied in following production, life and the scientific research.Meanwhile the simulation enzyme sensor of building is in the inspection of FKN
There is very strong specificity in survey.Using this key property of MOFs mimichydrogen peroxidase, the FKN sensor that we construct is
FKN detection in industrial production and medical diagnosis on disease provides more convenient and fast method.
Claims (4)
1. a kind of functional metal organic framework material, it is characterised in that the structure of the framework material are as follows: with Cu2+For center metal
Ion, 2- amino terephthalic acid (TPA) are that ligand has synthesized a kind of metal organic frame (MOFs) material under conditions of solvent heat
Cu(BDC-NH2)2, BDC expression p-phthalic bridge；The copper base metal skeleton Surface-modification of Nanoparticles has amino group.
2. a kind of FKN sensor is constructed using functional metal organic framework material according to claim 1, special
Sign is the sensor are as follows: knows using the fixed FKN antibody of the pSC-AuNPs self-assembled multilayer film in gold electrode surfaces as biology
Other surface；It is prepared for a kind of gold nanoparticle functionalization metal-organic framework materials of porosity sulfonic group calixarenes cladding simultaneously
H is catalyzed for signal label2O2。
3. a kind of method for preparing FKN sensor according to claim 2, it is characterised in that the specific steps of this method
Are as follows:
It a. is 10 by 2.8mL concentration-2Mol/L pSC and concentration are 2.63 × 10-2Mol/L gold chloride (HAuCl4) it is protected from light stirring
20min；Being rapidly added 2.8mL concentration later is 10-2Sodium borohydride (the NaBH of mol/L4), be stirred to react to the color of solution by
The faint yellow dark brown that becomes eventually becomes claret, stops reaction, is protected from light standing after 5-8 hours the pSC-AuNPs prepared
Be transferred in brown bottle be stored in 4 DEG C it is spare；
B. 100 μ L pSC-AuNPs are instilled into gained MOFs stirring 10-12 hours to prepare pSC-AuNPs/MOFs, then will
200μL 1μM NH2The FKN Apt of sealing end is added in above-mentioned pSC-AuNPs/MOFs solution, after stirring 10-12 hours
The Apt/pSC-AuNPs/MOFs of acquisition is modified to the three-dimensional electrode surface constructed in three-dimensional gold nano, utilizes MOFs hydrogen peroxide
The effect of analogue enztme is catalyzed H2O2Decomposition achieve the purpose that detect FKN.
4. according to the method described in claim 3, it is characterized in that the metal organic frame the preparation method comprises the following steps: stirring
Under, by polyvinylpyrrolidone, 2- amino terephthalic acid (TPA) and Cu (NO3)2·3H2O is dissolved in N, N- by the molar ratio of 1:3:10
Dimethylformamide and ethyl alcohol are mixed well, are heated 4-6 hours at 100 DEG C as in the volume ratio mixture of 1:1；On removing
Clear liquid, then be added DMF to dissolve gained sediment, by the solution 100 DEG C heating 8-10 hours to remove unreacted examination
Agent, centrifuge separation obtain metal organic frame.
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| CN111234243A (en) * | 2019-12-31 | 2020-06-05 | 郑州轻工业大学 | Metal organic framework material, preparation method thereof and immunosensor |
| CN111234243B (en) * | 2019-12-31 | 2021-10-01 | 郑州轻工业大学 | Metal organic framework material, preparation method thereof and immunosensor |
| CN112730561A (en) * | 2020-12-17 | 2021-04-30 | 中山大学 | Electrochemical biosensor based on supermolecule host-guest recognition technology and preparation method and application thereof |
| CN113702464A (en) * | 2021-09-23 | 2021-11-26 | 上海大学 | P-tau detection immunosensor and preparation and application methods thereof |
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