CN107383371B - A kind of protein-imprinted polymer microballoon and its preparation and application based on quantum dot - Google Patents
A kind of protein-imprinted polymer microballoon and its preparation and application based on quantum dot Download PDFInfo
- Publication number
- CN107383371B CN107383371B CN201710610059.6A CN201710610059A CN107383371B CN 107383371 B CN107383371 B CN 107383371B CN 201710610059 A CN201710610059 A CN 201710610059A CN 107383371 B CN107383371 B CN 107383371B
- Authority
- CN
- China
- Prior art keywords
- added
- protein
- imprinted polymer
- quantum
- preparation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
- C08G73/00—Macromolecular compounds obtained by reactions forming a linkage containing nitrogen with or without oxygen or carbon in the main chain of the macromolecule, not provided for in groups C08G12/00 - C08G71/00
- C08G73/06—Polycondensates having nitrogen-containing heterocyclic rings in the main chain of the macromolecule
- C08G73/0666—Polycondensates containing five-membered rings, condensed with other rings, with nitrogen atoms as the only ring hetero atoms
- C08G73/0672—Polycondensates containing five-membered rings, condensed with other rings, with nitrogen atoms as the only ring hetero atoms with only one nitrogen atom in the ring
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/22—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising organic material
- B01J20/26—Synthetic macromolecular compounds
- B01J20/268—Polymers created by use of a template, e.g. molecularly imprinted polymers
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/28—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties
- B01J20/28014—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties characterised by their form
- B01J20/28016—Particle form
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J9/00—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof
- C08J9/26—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof by elimination of a solid phase from a macromolecular composition or article, e.g. leaching out
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08K—Use of inorganic or non-macromolecular organic substances as compounding ingredients
- C08K3/00—Use of inorganic substances as compounding ingredients
- C08K3/30—Sulfur-, selenium- or tellurium-containing compounds
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08K—Use of inorganic or non-macromolecular organic substances as compounding ingredients
- C08K9/00—Use of pretreated ingredients
- C08K9/04—Ingredients treated with organic substances
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/02—Use of particular materials as binders, particle coatings or suspension media therefor
- C09K11/025—Use of particular materials as binders, particle coatings or suspension media therefor non-luminescent particle coatings or suspension media
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/08—Luminescent, e.g. electroluminescent, chemiluminescent materials containing inorganic luminescent materials
- C09K11/88—Luminescent, e.g. electroluminescent, chemiluminescent materials containing inorganic luminescent materials containing selenium, tellurium or unspecified chalcogen elements
- C09K11/881—Chalcogenides
- C09K11/883—Chalcogenides with zinc or cadmium
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2201/00—Foams characterised by the foaming process
- C08J2201/04—Foams characterised by the foaming process characterised by the elimination of a liquid or solid component, e.g. precipitation, leaching out, evaporation
- C08J2201/042—Elimination of an organic solid phase
- C08J2201/0424—Elimination of an organic solid phase containing halogen, nitrogen, sulphur or phosphorus atoms
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2379/00—Characterised by the use of macromolecular compounds obtained by reactions forming in the main chain of the macromolecule a linkage containing nitrogen with or without oxygen, or carbon only, not provided for in groups C08J2361/00 - C08J2377/00
- C08J2379/04—Polycondensates having nitrogen-containing heterocyclic rings in the main chain; Polyhydrazides; Polyamide acids or similar polyimide precursors
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
- G01N2021/6432—Quenching
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Polymers & Plastics (AREA)
- Analytical Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Materials Engineering (AREA)
- Physics & Mathematics (AREA)
- Immunology (AREA)
- Inorganic Chemistry (AREA)
- Optics & Photonics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention belongs to Materials Science and Engineering and bio-separation engineering field, a kind of specifically protein-imprinted polymer microballoon and its preparation and application based on fluorescence quantum.The microballoon with thioacetic acid (TGA) and glutathione (GSH) collectively as stabilizer CdTe quantum directly as function monomer, protein is template molecule, dopamine is crosslinking agent, obtains imprinted polymer microballoon by the auto polymerization of dopamine.The present invention is detected using the fluorescence signal of quantum dot, with the quantum dot of functionalization directly as function monomer, be ensure that and is all contained quantum dot in imprinted cavity, can be improved the sensitivity of detection.Meanwhile trace poly layer is obtained by the auto polymerization of dopamine, also further simplify printing process.For the analysis detection of bovine hemoglobin in phycocyanin in seawater and lake water sample, ox urine sample, recovery of standard addition can reach 90% or more, and repeat performance is good.
Description
Technical field
It is specifically a kind of based on quantum dot the invention belongs to Materials Science and Engineering and bio-separation engineering field
Protein-imprinted polymer microballoon and its preparation and application.
Background technique
Molecularly imprinted polymer is because its is highly selective, preparation is simple, anti-adverse environment ability is strong, stability is good, at low cost
The advantages that, it is developed rapidly in fields such as environment, life.Trace object also gradually develops from small molecule compound, ion
To large biological molecules such as protein, virus, cells.Small molecule compound and ion blotting are developed well at present,
But the trace of large biological molecule trace, especially protein is still the very challenging research neck in molecular engram field
Domain, main cause are: 1) protein molecule volume is big, is difficult the recognition site that disengaging is located at highly cross-linked polymeric inner, makes
Elution and the identification for obtaining protein molecule are difficult；2) protein molecular structure is complicated, conformation is flexible, it is easy to be denaturalized；3)
Protein is soluble easily in water, insoluble in conventional organic trace solvent or denaturation, due to the presence of water can destroy template and
The formation of hydrogen bond between monomer, so that imprinting effect is deteriorated.Therefore, effective and general western blotting method still right and wrong are explored
Normal stern challenge.
Currently, it constructs fluorescence sense system trace protein and other biological macromolecular is already a kind of feasible method.
In numerous fluorescent materials, quantum dot is obtained because of advantages such as luminous efficiency high, optics is adjustable, excites scope is wide, good light stabilities
To extensive use.However since quantum dot is selectively poor, if being directly applied to the analysis detection of actual complex sample,
Selectivity will become the critical issue of practical application.Therefore, by the height of the fluorescent characteristic of quantum dot and molecularly imprinted polymer
Selectivity combines, and preparing molecular engram fluorescent optical sensor will be in the identification and detection of target molecule with apparent excellent
Gesture.Quantum dot and molecular engram are combined into building molecular engram fluorescent optical sensor in recent years, and are applied to the selection of protein
Property identification and detection caused quite to pay close attention to, this kind of fluorescence imprinted material can simulate nature antigen-antibody identification process, will
Identification signal is converted to fluorescence signal convenient for capturing and analyzing.Quantum dot, molecular engram and Western blotting are combined into building
The research of fluorescence biomimetic sensor was having more report in recent years.For example, etc. developed series based on CdTe quantum
Molecular engram fluorescent optical sensor is for the red detection with bovine serum albumin of ox blood.Tang etc. reports a kind of fluorescence nano sensor use
In detection glycoprotein.Currently, fluorescent optical sensor of the building based on quantum dot is most or quantum dot is embedded in titanium dioxide silicon grain
Son or modify on the surface of silicon dioxide granule, then passes through free radical polymerization or sol-gel process in dioxy
SiClx surface prepares molecular engram layer, on the one hand these methods are difficult to ensure that quantum dot is equably embedded in highly cross-linked print
In mark polymer, comparatively laborious modification is in addition needed mostly, it is not only time-consuming but also uncontrollable.Therefore, it is badly in need of development
Simply, the method quickly and easily operated come prepare the molecular engram fluorescent optical sensor based on quantum dot for protein inspection
It surveys.
Summary of the invention
The protein-imprinted polymer microballoon and its preparation and answer that the purpose of the present invention is to provide a kind of based on quantum dot
With.
To achieve the above object, the technical solution adopted by the present invention are as follows:
A kind of protein-imprinted polymer microballoon based on quantum dot, polymer microballoon is with thioacetic acid (TGA) and paddy Guang
Sweet peptide (GSH) collectively as stabilizer CdTe quantum directly as function monomer, protein is template molecule, and dopamine is
Crosslinking agent obtains polymer microballoon by the auto polymerization of dopamine.
The protein is phycocyanin (PC) or bovine hemoglobin.
The CdTe quantum of the thioacetic acid and glutathione jointly stabilizing:
1) mixed liquor of second alcohol and water is added in tellurium powder and sodium borohydride after mixing, and then heats in 30-50 DEG C of water-bath anti-
It should be completely disappeared to the tellurium powder of black, until supernatant becomes lavender；
2) cadmium nitrate is added into excessive water, and thioacetic acid and glutathione is then added, and then regulation system pH is extremely
8-10 is added the supernatant that step 1) obtains tellurium powder product, flows back under nitrogen protection, obtain green thioacetic acid and paddy after adjusting
The CdTe quantum liquid of the sweet peptide jointly stabilizing of Guang.
A kind of preparation of the protein-imprinted polymer microballoon based on quantum dot, with thioacetic acid (TGA) and glutathione
(GSH) collectively as the CdTe quantum of stabilizer directly as function monomer, protein is template molecule, and dopamine is crosslinking
Agent is fluorescence nano sensor by the protein-imprinted polymer that the auto polymerization of dopamine obtains.
Specifically:
A. the preparation of thioacetic acid and the CdTe quantum of glutathione (TGA-GSH) jointly stabilizing: the thioacetic acid
With the CdTe quantum of glutathione jointly stabilizing:
1) mixed liquor of second alcohol and water is added in tellurium powder and sodium borohydride after mixing, and then 30-50 DEG C of constant temperature is anti-in a water bath
It should be completely disappeared to the tellurium powder of black, until supernatant becomes lavender；
2) cadmium nitrate is added into excessive water, and thioacetic acid and glutathione is then added, and then regulation system pH is extremely
8-10 is added the supernatant that step 1) obtains tellurium powder product, flows back under nitrogen protection, obtain green thioacetic acid and paddy after adjusting
The CdTe quantum liquid of the sweet peptide jointly stabilizing of Guang；For use；
B. the preparation of protein fluorescence nano print polymer: the quantum dot solution of above-mentioned TGA-GSH jointly stabilizing is taken, is added
Enter into phosphoric acid buffer (PBS) solution containing protein, dopamine is added after stirring, be added potassium peroxydisulfate (KPS), in water-bath
Reaction is overnight；Resulting polymer is washed with eluant ethanol/acetonitrile, to remove template protein, as Western blotting
Polymer.
The preparation of the step a. thioacetic acid and the CdTe quantum of glutathione jointly stabilizing: 1) 25-45mg is weighed
The ethyl alcohol of 0.5-2.5mL and the water of 0.1-1.5mL is added in the sodium borohydride of tellurium powder and 30-50mg, in 30-50 DEG C of water-bath
The tellurium powder of heating reaction to black completely disappears, until supernatant becomes lavender；
2) cadmium nitrate for weighing 80-100mg is added to 65-85mL water, and the thioacetic acid and 100- of 15-35 μ L is then added
120mg glutathione adjusts the pH of solution with the sodium hydroxide solution of 0.5-1.5M, blows 20-40min with nitrogen, then take step
The lavender supernatant 0.5-2mL of rapid 1) preparation is added in nitric acid cadmium solution, under nitrogen protection, is flowed back 1-3 hours, is obtained green
The CdTe quantum liquid of color thioacetic acid and glutathione jointly stabilizing.
The preparation of the step b. protein-imprinted polymer:
A) quantum dot solution for taking 4-6mL TGA-GSH jointly stabilizing is added to the 5- containing 20-40mg phycocyanin
In 20mL PBS (pH=6.5-8.5,5-20mM) solution, after stirring 2-6 hours, 20-40mg dopamine is added, nitrogen blows 10-
After 30min, 3-7mg KPS is added, 30-50 DEG C of water-soluble middle reaction is stayed overnight, by resulting polymer eluant ethanol/acetonitrile
(8:2, v/v) is washed 2-5 times, to remove removing template phycocyanin, by obtained phycocyanin imprinted polymer；
B) quantum dot solution for taking 4-6mL TGA-GSH jointly stabilizing is added to the 5- containing 5-20mg bovine hemoglobin
In 20mL PBS (pH=6.5-8.5,5-20mM) solution, after stirring 2-6 hours, 5-20mg dopamine is added, nitrogen blows 10-
After 30min, 3-7mg KPS is added, reaction is stayed overnight in 30-50 DEG C of aqueous solution, by resulting polymer eluant ethanol/acetonitrile
(8:2, v/v) is washed 2-5 times, to go removing template bovine hemoglobin, by obtained bovine hemoglobin imprinted polymer.
The western blot polymer, which is scattered in the ultrapure water of 0.1-2mL, to be saved, for use.
A kind of application of the protein-imprinted polymer microballoon based on quantum dot, the polymer microballoon is as fluorescence sense
Application of the device in detection protein.
Advantage for present invention:
The present invention is selection identification and highly sensitive inspection of the fluorescence nano trace sensor based on quantum dot for protein
It surveys, specifically Selective recognition and detection protein, protein further can be phycocyanin and bovine hemoglobin.By using
Thioacetic acid and glutathione collectively as stabilizer CdTe quantum directly as function monomer, effectively prevent cumbersome
Quantum dot surface modification, has also carried out it while stablizing quantum dot surface-functionalized, ensure that in imprinted cavity
All contain quantum dot, improves the sensitivity of detection.The thickness of gained fluorescence nano imprinted layer is about 3nm, so thin trace
Layer help to obtain higher detection sensitivity and shorter reaction time.Furthermore it is possible to which the fluorescent quenching by quantum dot is real
Now to the detection of phycocyanin and bovine hemoglobin, to improve detection sensitivity.Compared to other albumen, the imprinted polymer pair
Phycocyanin and bovine hemoglobin have higher identification selection, binding capacity and absorption stability.To seawater and lake water sample
Phycocyanin in product, in ox urine sample bovine hemoglobin analysis detection, recovery of standard addition can reach 90% or more, and repeat
Service performance is good.The present invention has both the advantages such as quick, high selection, highly sensitive, easy to operate, inexpensive, can be used as a hatching egg
White detection new strategy enriches molecular engram/protein correlative study.
Detailed description of the invention
Fig. 1 is the preparation process schematic diagram of phycocyanin imprinted polymer provided in an embodiment of the present invention.
Fig. 2 is transmission electron microscope picture provided in an embodiment of the present invention: A, CdTe QDs, B, phycocyanin fluorescence nano trace are poly-
It closes object (PC MIPs), C, non-imprinted polymer (PC NIPs), D, bovine hemoglobin fluorescence nano imprinted polymer (BHb
) and E, non-imprinted polymer (BHbNIPs) MIPs.
Fig. 3 is phycocyanin fluorescence nano imprinted polymer (PCMIPs) provided in an embodiment of the present invention and bovine hemoglobin
As pH changes after fluorescence intensity change under fluorescence nano imprinted polymer (BHb MIPs) difference pH, and absorption phycocyanin
Fluorescence intensity change ratio figure.
Fig. 4 is phycocyanin fluorescence nano imprinted polymer (PCMIPs) provided in an embodiment of the present invention and non-trace polymerization
Object (PC NIPs) and bovine hemoglobin fluorescence nano imprinted polymer (BHb MIPs) and non-imprinted polymer (BHb NIPs) with
Phycocyanin is added the amount of amount and bovine hemoglobin increase the variation diagram of its fluorescence maximum emission wavelength.
Fig. 5 is phycocyanin fluorescence nano imprinted polymer (PCMIPs) provided in an embodiment of the present invention and non-trace polymerization
Object (PC NIPs) and the red fluorescence nano imprinted polymer (BHbMIPs) of ox blood and non-imprinted polymer (BHb NIPs) are to identical
Concentration difference protein selectivity lab diagram.
Specific embodiment
With reference to the accompanying drawing and embodiment the present invention will be further explained explanation.
The method of the present invention is relatively simple building using quantum dot as the nano print sensor, method in fluorescence signal source: with
Thioacetic acid and glutathione collectively as stabilizer CdTe quantum directly as function monomer, phycocyanin or ox blood are red
Albumen is template molecule, and dopamine is crosslinking agent, prepares protein-imprinted polymer by the auto polymerization of dopamine.This method has
Effect avoid cumbersome quantum dot surface modification, be directly added into the synthesis process of quantum dot stabilizer thioacetic acid and
Glutathione, stablize quantum dot while also it has been carried out it is surface-functionalized, in addition, with the quantum dot of functionalization directly as
Function monomer ensure that and all contain quantum dot in imprinted cavity, can be improved the sensitivity of detection.Meanwhile passing through dopamine
Auto polymerization obtains trace poly layer, also further simplifies the process of trace.For phycocyanin, ox urine in seawater and lake water sample
The analysis detection of bovine hemoglobin in sample, recovery of standard addition can reach 90% or more, and repeat performance is good.
Embodiment 1
A. 38.3mg tellurium powder and 40mg the preparation of thioacetic acid and the CdTe quantum of glutathione jointly stabilizing: are weighed
The ethyl alcohol of 1.5mL and the water of 0.5mL is added in sodium borohydride, makes system isolating oxygen, and heating is reacted to black in 40 DEG C of water-bath
Tellurium powder completely disappear, until supernatant becomes lavender.The cadmium nitrate for weighing 92.4mg is added in 75mL water, is then added
The thioacetic acid and 110mg glutathione of 25 μ L adjusts pH to 9 with the sodium hydroxide solution of 1M, and nitrogen blows 30min, then takes
The lavender supernatant 1mL for stating preparation is added in nitric acid cadmium solution, under nitrogen protection, flows back 2 hours to get green sulfydryl second
The CdTe quantum liquid of acid and glutathione (TGA-GSH) jointly stabilizing.
B. the preparation of phycocyanin imprinted polymer: the quantum dot of the above-mentioned TGA-GSH jointly stabilizing prepared of 5mL is taken
Solution is added in 10mL PBS (pH=7.5,10mM) solution containing 30mg phycocyanin, after stirring 4 hours, is added
After nitrogen blows 20min, 5mg KPS is added, 40 DEG C of water-soluble middle reactions are overnight in 30mg dopamine.By resulting polymer eluent
Ethanol/acetonitrile (8:2, v/v) is washed 3 times, to remove removing template phycocyanin, is dispersed imprinted polymer obtained in again
It is saved in the ultrapure water of 0.5mL.As control, non-imprinted polymer (PCNIPs) is prepared using identical method, is only being printed
Template molecule phycocyanin is not added during mark.
The preparation of bovine hemoglobin imprinted polymer (BHb MIPs) uses method same as described above, only in trace mistake
The template molecule being added in journey changes bovine hemoglobin into, while the additional amount of bovine hemoglobin is 10mg, and the amount of dopamine is
10mg, the corresponding red non-imprinted polymer (BHbNIPs) of ox blood are added without template molecule in the preparation.
Embodiment 2
Take respectively 10 μ L through ultrapure water dilute 1000 times above-described embodiment obtain CdTe quantum, phycocyanin trace
Imprinted polymer (PC MIPs) and non-imprinted polymer (PC NIPs) and bovine hemoglobin imprinted polymer (BHb MIPs) and
Each dilution is dispersed on the copper mesh by ethyl alcohol cleaning by non-imprinted polymer (BHb NIPs) solution respectively, after dry,
The copper mesh for being loaded with substance after above-mentioned each dilution is observed into (A-E referring to fig. 2) with transmission electron microscope；As shown in Figure 2 A, quantum dot
Partial size be about 2nm, as shown in Figure 2 B, the partial size of PC MIPs is about 5nm, and dispersibility very well, phycocyanin imprinted layer
Thickness is about 3nm, and so thin imprinted layer is very favorable for obtaining higher detection sensitivity and faster reacting.
As shown in Figure 2 C, the partial size of PC NIPs and pattern and quantum dot is similar, illustrates in the case where phycocyanin is not present,
Dopamine is only merely to be modified quantum dot surface, has obtained PDA film, for its particle size not how many change
Become.As it can be seen that template molecule phycocyanin has played very big effect for the formation of imprinted layer in printing process.For BHb
MIPs and BHb NIPs, as shown in Fig. 2 D, E, pattern is similar with corresponding PC MIPs and PC NIPs pattern.
Embodiment 3
After phycocyanin imprinted polymer obtained (PC MIPs) solution ultrasonic disperse, parallel 3 parts, 1mL is taken respectively
PC MIPs solution is added in the 1.5mL centrifuge tube weighed, and claims to obtain quality after drying again.Before calculating 3 parts of parallel samples
Ratio of poor quality with volume afterwards, average value are the concentration of PC MIPs solution, concentration 4.6mg/mL.
The PC MIPs solution that the above-mentioned acquisition concentration of 0.2mL is 4.6mg/mL is taken when detection, measurement overall solution volume is
0.5mL, the i.e. concentration of PC MIPs are 1.84mg/mL.The red fluorescence nano trace of ox blood in detection process is obtained in the same way
The concentration of polymer (BHb MIPs) is 1mg/mL.
Secure ph is a series of PBS buffer solutions such as 6.0,6.5,7.0,7.5,8.0,8.5 and 9.0.First group each
Sample takes the solution of the phycocyanin imprinted polymer (PC MIPs) of 0.2mL, then be separately added into a series of difference pH PBS it is slow
Solution 0.3mL is rushed, then mixing oscillation measures the fluorescence intensity of each sample with luminoscope.Second group of each sample takes 0.2mL
PCMIPs solution and 0.01mL 0.2mM phycocyanin solution (concentration of i.e. surveyed phycocyanin be 4 μM), respectively plus
Enter the PBS buffer solution 0.29mL of difference pH of above-mentioned configuration a series of, mixing oscillation a period of time, is then measured with luminoscope
The fluorescence intensity of each sample.The experimental method of bovine hemoglobin imprinted polymer (BHb MIPs) is similar to above, surveyed ox
The concentration of hemoglobin is 2 μM.
As shown in Figure 3A, for phycocyanin imprinted polymer (PC MIPs), when pH is less than 6.5, fluorescence enhancement efficiency
Value increases with the increase of pH；When pH is greater than 6.5, with the increase of pH value, fluorescent quenching efficiency value can reduce, experimental result table
Bright, the optimal pH for detecting phycocyanin is 6.5.Shown in Fig. 3 B, for bovine hemoglobin imprinted polymer (BHb MIPs),
When pH is less than 7, fluorescence enhancement efficiency value increases with the increase of pH；When pH is greater than 7, with the increase of pH value, fluorescent quenching effect
Rate value can reduce, the experimental results showed that, the optimal pH for detecting bovine hemoglobin is 7, under conditions of weakly acidic pH, studies albumen
Matter has potential application.
Embodiment 4
It is similar to method in embodiment 3, measure phycocyanin imprinted polymer (PC MIPs) and non-imprinted polymer (PC
NIPs concentration) is 1.84mg/mL.Then, the PC NIPs and pH of the PC MIPs0.2mL or above-mentioned concentration that take above-mentioned concentration be
Two groups of samples of above-mentioned acquisition are separately added into a series of various concentrations of 0.01mL by the mixed liquor 0.29mL of 6.5 PBS buffer solution
Phycocyanin solution (ultimate density of phycocyanin is respectively 0.8,1.2,1.6,2,2.4,3,3.6,4,5,6,7,8 μM),
Mixing oscillation a period of time, the fluorescence intensity of each sample is then measured with luminoscope.
Bovine hemoglobin imprinted polymer (BHb MIPs) is similar with the detection method of non-imprinted polymer (BHb NIPs),
The concentration of BHb MIPs and BHb NIPs are that 1mg/mL then takes the BHb MIPs 0.2mL or above-mentioned concentration BHb of above-mentioned concentration
It is a series of to be separately added into 0.01mL by the mixed liquor 0.29mL for the PBS buffer solution that NIPs and pH is 7 for two groups of samples of above-mentioned acquisition
Various concentration bovine hemoglobin solution (ultimate density of bovine hemoglobin be 0.3,0.5,0.75,1,1.5,2,2.5,3,
3.5,4,4.5,5 μM), mixing oscillation a period of time, the fluorescence intensity of each sample is then measured with luminoscope.
As shown in Figure 4 A, due to the increase with phycocyanin concentration, the emission peak intensity of quantum dot dies down, and algae indigo plant egg
White emission peak enhancing directly results in fluorescence color and is gradually transitions by previous green orange, ultimately becomes red, Neng Gouyong
Naked eyes see differentiable, apparent color change, thus Visual retrieval phycocyanin.As shown in Figure 4 B, for PC
NIPs, with the increase of phycocyanin concentration, the emission peak of the quantum dot at 545nm is declined slightly, and the range of linearity is very
It is narrow, illustrate that non-imprinted polymer is very low to the sensitivity of phycocyanin, it is such the experimental results showed that imprinted polymer can be shown
The quenching efficiency for improving quantum dot is write, to improve the detection sensitivity of phycocyanin, this is because imprinted polymer has pair
Caused by the specific recognition site of phycocyanin.For BHb MIPs and NIPs, it may have similar experimental result, such as Fig. 4 C, D institute
Show, with the increase of bovine hemoglobin concentration, the emission peak intensity of quantum dot is also with reduction, rather than imprinted polymer is to ox blood
The sensitivity of Lactoferrin is very low.It is above-mentioned the experimental results showed that two kinds of imprinted polymers than corresponding non-imprinted polymer all to respective
Template protein show very high sensitivity.
Embodiment 5
It is similar to the above method, phycocyanin imprinted polymer (PC MIPs) and non-imprinted polymer (PC NIPs) it is dense
Degree is 1.84mg/mL, then, the PBS that the PC NIPs and pH of the PC MIPs 0.2mL or above-mentioned concentration that take above-mentioned concentration are 6.5
The mixed liquor 0.29mL of buffer solution, by two groups of samples of above-mentioned acquisition be separately added into 0.01mL concentration be 34g/L phycocyanin,
Phycoerythrin or spirulina solution, that is, the concentration of surveyed different albumen are 680mg/L, mixing oscillation a period of time, then with glimmering
Light instrument measures the fluorescence intensity of each sample.
Bovine hemoglobin imprinted polymer (BHb MIPs) is similar with the detection method of non-imprinted polymer (BHb NIPs),
The concentration of BHb MIPs and BHb NIPs be 0.1mg/mL, then, take above-mentioned concentration BHb MIPs 0.2mL or above-mentioned concentration
The mixed liquor 0.29mL for the PBS buffer solution that BHb NIPs and pH are 7, it is equal by concentration is separately added into two groups of samples of above-mentioned acquisition
For 2.5 μM of bovine hemoglobin, bovine serum albumin, lysozyme or phycocyanin solution, mixing oscillation a period of time, then with glimmering
Light instrument measures the fluorescence intensity of each sample.
As shown in Figure 5A, template molecule phycocyanin is maximum to the quenching degree of quantum dot.Followed by spirulina powder, this is
Because containing part phycocyanin in spirulina powder, there is certain quenching effect to quantum dot, phycoerythrin is because structure is with algae indigo plant
Albumen is more close, and quenching degree is more slightly lower than spirulina.Meanwhile above-mentioned albumen is suitable to the quenching degree of non-trace microballoon,
This is because the recognition site that non-imprinted polymer surface is not specific, the absorption to albumen is only non-specific adsorption.
Similarly, bovine hemoglobin is much higher than cow's serum, lysozyme and phycocyanin (Fig. 5 B) to the quenching degree of quantum dot, this be because
Contain for BHb MIPs largely to the specific recognition site of bovine hemoglobin, other albumen are because of configuration, size etc. and recognition site
It mismatches, thus hardly enters imprinted cavity, so that quenching degree is lower.In addition, phycocyanin also have to BHb MIPs it is very low
Quenching efficiency, thus more demonstrate imprinted polymer only to template protein have very high selectivity.
Claims (8)
1. a kind of protein-imprinted polymer microballoon based on fluorescence quantum, it is characterised in that: polymer microballoon is with sulfydryl second
Sour (TGA) and glutathione (GSH) collectively as stabilizer CdTe quantum directly as function monomer, protein is template
Molecule, dopamine are crosslinking agent, obtain polymer microballoon by the auto polymerization of dopamine.
2. the protein-imprinted polymer microballoon according to claim 1 based on fluorescence quantum, it is characterised in that: the egg
White matter is phycocyanin (PC) or bovine hemoglobin (BHb).
3. the protein-imprinted polymer microballoon as described in claim 1 or 2 based on fluorescence quantum, it is characterised in that: institute
State the preparation method of the CdTe quantum of thioacetic acid and glutathione jointly stabilizing:
1) mixed liquor of second alcohol and water is added in tellurium powder and sodium borohydride after mixing, and then heating is reacted extremely in 30-50 DEG C of water-bath
The tellurium powder of black completely disappears, until supernatant becomes lavender；
2) cadmium nitrate is added into excessive water, thioacetic acid and glutathione is then added, then regulation system pH to 8-10,
The supernatant that step 1) obtains tellurium powder product is added after adjusting, flows back under nitrogen protection, obtains green thioacetic acid and gluathione
The CdTe quantum liquid of peptide jointly stabilizing.
4. a kind of preparation method of the protein-imprinted polymer microballoon described in claim 1 based on fluorescence quantum, special
Sign is: with thioacetic acid (TGA) and glutathione (GSH) collectively as stabilizer CdTe quantum directly as function list
Body, protein are template molecule, and dopamine is crosslinking agent, and the protein-imprinted polymer obtained by the auto polymerization of dopamine is micro-
Ball is fluorescence nano trace sensor；
Specially
A. the preparation of thioacetic acid and the CdTe quantum of glutathione (TGA-GSH) jointly stabilizing:
1) mixed liquor of second alcohol and water is added after tellurium powder and sodium borohydride mixing, then 30-50 DEG C of isothermal reaction be extremely in a water bath
The tellurium powder of black completely disappears, until supernatant becomes lavender；
2) cadmium nitrate is added into excessive water, thioacetic acid and glutathione is then added, then regulation system pH to 8-10,
The supernatant that step 1) obtains tellurium powder product is added after adjusting, flows back under nitrogen protection, obtains green thioacetic acid and gluathione
The CdTe quantum liquid of peptide jointly stabilizing；For use；
B. the preparation of protein nano imprinted polymer: the quantum dot solution of above-mentioned TGA-GSH jointly stabilizing is taken, is added to and contains
Have in phosphoric acid buffer (PBS) solution of protein, dopamine is added after stirring, be added potassium peroxydisulfate (KPS), was reacted in water-bath
Night；Resulting polymer is washed with eluant ethanol/acetonitrile, to remove template protein, as protein-imprinted polymer.
5. the preparation method of the protein-imprinted polymer microballoon according to claim 4 based on fluorescence quantum, feature
It is: the preparation of the step a. thioacetic acid and the CdTe quantum of glutathione jointly stabilizing: 1) weighs 25-45 mg tellurium
The ethyl alcohol of 0.5-2.5 mL and the water of 0.1-1.5 mL is added, in 30-50 DEG C of water-bath in the sodium borohydride of powder and 30-50 mg
The tellurium powder of middle heating reaction to black completely disappears, until supernatant becomes lavender；
2) cadmium nitrate for weighing 80-100 mg is added to 65-85 mL water, and the thioacetic acid and 100- of 15-35 μ L is then added
120 mg glutathione adjust the pH of solution with the sodium hydroxide solution of 0.5-1.5 M, blow 20-40 min with nitrogen, then take
The lavender supernatant 0.5-2 mL of step 1) preparation is added in nitric acid cadmium solution, under nitrogen protection, is flowed back 1-3 hours, is obtained
To the CdTe quantum liquid of green thioacetic acid and glutathione jointly stabilizing.
6. the preparation method of the protein-imprinted polymer microballoon according to claim 4 based on fluorescence quantum, feature
It is: the preparation of the step b. protein fluorescence nano print polymer:
A) quantum dot solution for taking 4-6 mL TGA-GSH jointly stabilizing, is added to the 5-20 containing 20-40 mg phycocyanin
In mL pH=6.5-8.5,5-20 mM PBS solution, after stirring 2-6 hours, 20-40 mg dopamine is added, nitrogen blows 10-30
After min, 3-7 mg KPS is added, resulting polymer overnight, is by 30-50 DEG C of water-soluble middle reaction with eluant ethanol/acetonitrile
V/v=8:2 is washed 2-5 times, to remove removing template phycocyanin, by obtained phycocyanin imprinted polymer；
B) quantum dot solution for taking 4-6 mL TGA-GSH jointly stabilizing, is added to the 5-20 containing 5-20 mg bovine hemoglobin
In mL pH=6.5-8.5,5-20 mM PBS solution, after stirring 2-6 hours, 5-20 mg dopamine is added, nitrogen blows 10-30
After min, 3-7 mg KPS is added, reaction is stayed overnight in 30-50 DEG C of aqueous solution, by resulting polymer eluant ethanol/acetonitrile
For v/v=8:2, wash 2-5 times, to go removing template bovine hemoglobin, by obtained bovine hemoglobin imprinted polymer.
7. the preparation method of the protein-imprinted polymer microballoon according to claim 6 based on fluorescence quantum, feature
Be: the western blot polymer, which is scattered in the ultrapure water of 0.1-2 mL, to be saved, for use.
8. a kind of application of the protein-imprinted polymer microballoon described in claim 1 based on fluorescence quantum, feature exist
In: application of the polymer microballoon as fluorescence nano trace sensor in detection protein.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710610059.6A CN107383371B (en) | 2017-07-25 | 2017-07-25 | A kind of protein-imprinted polymer microballoon and its preparation and application based on quantum dot |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710610059.6A CN107383371B (en) | 2017-07-25 | 2017-07-25 | A kind of protein-imprinted polymer microballoon and its preparation and application based on quantum dot |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107383371A CN107383371A (en) | 2017-11-24 |
| CN107383371B true CN107383371B (en) | 2019-09-27 |
Family
ID=60336836
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710610059.6A Active CN107383371B (en) | 2017-07-25 | 2017-07-25 | A kind of protein-imprinted polymer microballoon and its preparation and application based on quantum dot |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107383371B (en) |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107964065B (en) * | 2017-11-29 | 2020-04-24 | 中国科学院烟台海岸带研究所 | Temperature-sensitive type western blot sensing microsphere based on quantum dots and preparation method thereof |
| CN109655442B (en) * | 2019-03-14 | 2019-06-11 | 中国科学院烟台海岸带研究所 | Three fluorescent emissive molecules trace sensors of one kind and its preparation and application |
| CN111534299B (en) * | 2020-04-29 | 2023-06-27 | 上海应用技术大学 | GOQDs@PDA-ir-MIP molecularly imprinted fluorescent sensor and preparation method and application thereof |
| CN111829999B (en) * | 2020-07-23 | 2021-06-25 | 重庆大学 | Application method of perovskite fluorescent microsphere and dopamine system |
| CN113189064B (en) * | 2021-04-22 | 2022-12-09 | 西安交通大学 | Glycopeptide-based fluorescent molecularly imprinted polymer, preparation method and application in screening and detecting glycoprotein |
| CN117304491B (en) * | 2023-10-25 | 2024-03-29 | 宁夏医科大学 | Hemoglobin molecularly imprinted polymer based on silicon emulsion self-assembly and preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102313725A (en) * | 2011-07-21 | 2012-01-11 | 南开大学 | Preparation method of lysozyme molecular imprinting-quantum dot nanoscale fluorescent probe |
| CN104075925A (en) * | 2014-07-22 | 2014-10-01 | 中国科学院烟台海岸带研究所 | Preparation method for quantum dot-based protein core-shell imprinted microspheres |
| CN104165874A (en) * | 2014-07-24 | 2014-11-26 | 江苏大学 | Quantum dot fluorescent aspirin imprinted sensor and its preparation method and use |
-
2017
- 2017-07-25 CN CN201710610059.6A patent/CN107383371B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102313725A (en) * | 2011-07-21 | 2012-01-11 | 南开大学 | Preparation method of lysozyme molecular imprinting-quantum dot nanoscale fluorescent probe |
| CN104075925A (en) * | 2014-07-22 | 2014-10-01 | 中国科学院烟台海岸带研究所 | Preparation method for quantum dot-based protein core-shell imprinted microspheres |
| CN104165874A (en) * | 2014-07-24 | 2014-11-26 | 江苏大学 | Quantum dot fluorescent aspirin imprinted sensor and its preparation method and use |
Non-Patent Citations (2)
| Title |
|---|
| Quantum dots based imprinting fluorescent nanosensor for the selective and sensitive detection of phycocyanin: A general imprinting strategy toward proteins;xiaoyan Wang;《Sensors and Actuators B》;20170810;第255卷;268-274 * |
| 磁纳米分子印迹聚合物的制备及性能;刘微波等;《食品科学》;20111231;第32卷(第10期);61-65 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107383371A (en) | 2017-11-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107383371B (en) | A kind of protein-imprinted polymer microballoon and its preparation and application based on quantum dot | |
| Ansari et al. | Recent advances and future trends on molecularly imprinted polymer-based fluorescence sensors with luminescent carbon dots | |
| Jalili et al. | Detection of penicillin G residues in milk based on dual-emission carbon dots and molecularly imprinted polymers | |
| Yang et al. | Strategies of molecular imprinting-based fluorescence sensors for chemical and biological analysis | |
| Wang et al. | Molecular imprinting based hybrid ratiometric fluorescence sensor for the visual determination of bovine hemoglobin | |
| Zhang et al. | Quantum dots based mesoporous structured imprinting microspheres for the sensitive fluorescent detection of phycocyanin | |
| Wu et al. | Molecularly imprinted polymers-coated gold nanoclusters for fluorescent detection of bisphenol A | |
| JP5400297B2 (en) | Fluorescent nanosilica particles, nanofluorescent material, biochip using the same, and assay method thereof | |
| Chullasat et al. | Nanocomposite optosensor of dual quantum dot fluorescence probes for simultaneous detection of cephalexin and ceftriaxone | |
| CN110082328A (en) | A kind of carbon quantum dot-molecular engram fluorescent sensor material and its preparation method and application detecting chloramphenicol | |
| Piloto et al. | Label-free quantum dot conjugates for human protein IL-2 based on molecularly imprinted polymers | |
| Zhao et al. | Silanized carbon dot-based thermo-sensitive molecularly imprinted fluorescent sensor for bovine hemoglobin detection | |
| Wei et al. | Fabrication and evaluation of sulfanilamide-imprinted composite sensors by developing a custom-tailored strategy | |
| Jiang et al. | Surface molecular imprinting on CdTe quantum dots for fluorescence sensing of 4-nitrophenol | |
| CN103937486B (en) | A kind of fluorescent nano probe and its preparation method and application | |
| Zhao et al. | Synthesis of multi-mode quantum dots encoded molecularly imprinted polymers microspheres and application in quantitative detection for dopamine | |
| Lu et al. | Surface molecular imprinting on silica-coated CdTe quantum dots for selective and sensitive fluorescence detection of p-aminophenol in water | |
| Xiao et al. | Fluorescent nanomaterials combined with molecular imprinting polymer: synthesis, analytical applications, and challenges | |
| Wei et al. | A novel molecularly imprinted polymer thin film at surface of ZnO nanorods for selective fluorescence detection of para-nitrophenol | |
| CN106680496A (en) | Preparation of colorimetric and fluorescent double-signal nanospheres and application of colorimetric and fluorescent double-signal nanospheres to immunochromatographic quantitative detection | |
| CN107325239B (en) | A kind of high specific sulfanilamide (SN) trace quantum dot fluorescence sensor and preparation method thereof | |
| Wang et al. | Novel strategy to prepare fluorescent polymeric nanoparticles based on aggregation-induced emission via precipitation polymerization for fluorescent lateral flow assay | |
| CN109490523A (en) | Method for the nano material of label, nucleic acid probe and nucleic acid and nano material coupling | |
| Zhang et al. | Surface-imprinted polymer coating l-cysteine-capped ZnS quantum dots for target protein specific recognition | |
| Wang et al. | Fluorescent molecularly imprinted nanoparticles with boronate affinity for selective glycoprotein detection |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |