CN102814153A - Preparation and application of composite structure microsphere with color change ability - Google Patents
Preparation and application of composite structure microsphere with color change ability Download PDFInfo
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Abstract
The invention discloses a composite microsphere with a biosensing effect and its preparation method. The preparation method of the composite microsphere comprises: using polyelectrolyte as a molecular glue, by means of static electricity and hydrophobic interaction, assembling diacetylene vesicles on the surface of a polystyrene microsphere, thus forming a composite microsphere particle; and subjecting the composite microsphere particle to ultraviolet irradiation, thus obtaining the polystyrene/polydiacetylene vesicle composite microsphere with color change and biomolecule recognition abilities. The microsphere utilizes the polyelectrolyte as the molecular glue to make diacetylene adsorbed onto the microsphere surface. Compared with other forms of diacetylene sensors, the microsphere provided in the invention has the advantages of stability, easy storage, easy separation, and strong anti-interference property. Compared with enzyme-linked immunosorbent assay and other immunodetection methods, the microsphere also has the characteristics of simplicity, rapidity, visibility to naked eyes, and low price, etc. At the same time, the composite microsphere can also be used in micro-reactors and used for separation, drug carriers as well as various aspects.
Description
Technical field
The present invention relates to a kind of preparation and application thereof with composite construction microballoon of metachrosis.
Background technology
Gathering diacetylene (PDA) biology sensor has just got more and more people's extensive concerning since it comes out.Gather diacetylene and have unique conjugated structure, under ultraviolet lighting, can cause the transition of delocalized, photopolymerization takes place; Present bright blueness, when the skeleton that gathers diacetylene when blueness received the interference of environmental stimuli, the conformation of polymer backbone can change; Cause its absworption peak blue shift; Make the color transition that gathers diacetylene for red, and produce unique fluorescent characteristic, thereby realize detection stimulus.The PDA biology sensor is for solvent, pH value, temperature, biomolecule (sugar-protein; Ag-Ab; DNA etc.) has recognition reaction; The perception environmental stimuli produces macroscopic response signal fast, and can carry out detection by quantitative through ultraviolet-visible absorption spectroscopy or fluorescence microscope.
The PDA biology sensor is expansion gradually in its evolution, and its traditional device comprises: the LB/LS film, and vesica solution, fiber etc., but these several kinds of forms all have many defectives.For example there is complicated operation in the LB/LS film, uses problems such as not convenient, and vesica solution is not easy to separate; Break easily; And fiber is not suitable for biomolecule identification field, and these drawbacks have seriously restricted the development of PDA biology sensor, how to make the PDA sensor both portable; Stable, have the focus that recognition reaction has widely become research again.
Bibliographical information was once arranged; Utilize the method for layer assembly the vesica solution of PDA to be attached to the surface of substrate of glass; Thereby increase its storage capacity and portable performance, yet the interference of glass itself has bigger influence to identification, simultaneously; Fixing PDA sensor is not easy to combine effectively with biomolecule, and then has limited its identification range and ability.In order to increase the portability of sensor; The effect that someone research and utilization electric charge adsorbs is adsorbed on PDA on the surface of silica spheres; Though can carry out bio-identification qualitatively, because a little less than the simple electrostatic adsorption power, and have the saturated problem of absorption; How to improve the photopolymerization performance of PDA, become difficult point thereby improve its detection performance.
Along with the development of society, the factor that influences people's existence and life is more and more: volatile organic solvent, heavy metal ion; Bacterium, virus etc., these life and healths that threatening us at all times; In order to find as early as possible and to carry out preventing and controlling, a kind of simple, portable; Stable, the appearance that can be used for the trace detection sensor becomes the urgent demand of people.
Summary of the invention
One of the object of the invention provides the preparation method of a kind of polystyrene/diacetylene vesica complex microsphere.
The method for preparing polystyrene/diacetylene vesica complex microsphere provided by the present invention comprises the steps:
1) preparation diacetylene (PCDA) vesica solution;
2) adopt the static self-assembly method at charged polystyrene microsphere surface deposition polyelectrolyte layer, obtain coating the polystyrene microsphere (being the polystyrene polyelectrolyte microsphere) of polyelectrolyte layer; Wherein, said polyelectrolyte layer is at least one deck, and outermost polyelectrolyte is positively charged;
3) in step 2) diacetylene (PCDA) vesica of the polystyrene microsphere surface adsorption step 1) preparation of the coating polyelectrolyte layer that obtains, obtain said polystyrene/diacetylene vesica complex microsphere (polystyrene polyelectrolyte diacetylene microballoon).
Contain more PCDA vesica in the said composite construction microballoon in order to make; Said method comprises the steps: that also the composite construction microsphere surface that obtains in step 3) adsorbs cationic polyelectrolyte earlier; And then absorption diacetylene (PCDA) vesica, this step can repeat 0-2 time, preferably repeats 2 times.
Above-mentioned steps 1) in, said diacetylene vesica solution specifically can prepare according to following method: the diacetylene monomer powders is added in the phosphate buffer solution, and ultrasonic being uniformly dispersed obtains diacetylene solution; Said diacetylene solution was placed 512 hours 0-4 ℃ of following lucifuge, and using the aperture then is that the miillpore filter of 0.2 μ m-0.5 μ m filters, and promptly gets diacetylene vesica solution and stores for future use.The ultrasonic cell pulverization appearance of said ultrasonic concrete SCIENIZ JY92-II type capable of using carries out.
This preparation method is with respect to the preparation of traditional diacetylene vesica solution; Have simple to operate, stable, characteristics such as good reproducibility; In diacetylene vesica preparation is beyond example, and this lays a good foundation for the preparation of follow-up composite and the stability of keeping this material.
Above-mentioned steps 2) in, said polystyrene microsphere can obtain through commercial sources, and the microballoon size can be 100nm-1 μ m.Its surperficial positively chargeable or negative electrical charge.According to the charged difference in polystyrene microsphere surface, select the polyelectrolyte opposite to deposit with its charging property.
Step 2) polyelectrolyte layer that is deposited in can be single or multiple lift.When said polyelectrolyte layer was multilayer, it was the sequence layer that is made up of cationic polyelectrolyte and anionic polyelectrolyte alternating deposit, and its sequence number of plies is unrestricted.ABA for example ... A or BA ... A, wherein, A is a cationic polyelectrolyte, B is an anionic polyelectrolyte.Usually for surface of stability electric charge, obtain stable structure more, said polyelectrolyte layer is selected deposition 3-6 layer.
Said cationic polyelectrolyte can be polymine (PEI) or PAH hydrochloride (PAH).
Said anionic polyelectrolyte can be kayexalate (PSS), polystyrene sodium sulphate (PAA) or polyacrylic acid (PAA).
For on polystyrene microsphere surface-assembled diacetylene vesica (surface electronegative), so step 2) the polystyrene polyelectrolyte microsphere surface that obtains is all positively charged.
With the electronegative polystyrene microsphere in surface is example; To step 2) the preparation detailed method of polystyrene polyelectrolyte microsphere that has positive electricity describes: it is electronegative that the polystyrene microsphere surface of being adopted has sulfonic group; In its aqueous solution, add cationic polyelectrolyte; Leave standstill hatching 15-20min, repeatedly centrifugally remove unnecessary polyelectrolyte, obtain depositing the solution of the polystyrene polyelectrolyte microsphere of one deck polyelectrolyte (positively charged).
In order to obtain the polystyrene microsphere that multi-layer polyelectrolyte coats, can in said solution, add anionic polyelectrolyte, leave standstill 15-30min; Repeatedly centrifugally remove unnecessary anionic polyelectrolyte; Then add cationic polyelectrolyte again, leave standstill 15-30min, repeatedly centrifugally remove unnecessary cationic polyelectrolyte; Repeat aforesaid operations several times (as 0-4 time) as required, obtain the polystyrene polyelectrolyte microsphere solution of positively charged.
If said cationic polyelectrolyte is a polymine, needing the pH value of regulator solution before adding is 3-3.2; If said cationic polyelectrolyte is the PAH hydrochloride, needing the pH value of regulator solution before adding is 7.5; Adding anionic polyelectrolyte preceding need regulator solution pH value be 3.3-3.5.
The detailed method of preparation polystyrene polyelectrolyte diacetylene composite construction microballoon is following in the step 3): described diacetylene vesica solution is joined in the polystyrene polyelectrolyte microsphere solution of positively charged; Standing adsorption 20-60min; Repeatedly centrifugally remove the diacetylene vesica that not absorption is got on, obtain electronegative polystyrene polyelectrolyte diacetylene microspheres solution.
Contain more PCDA vesica in the said composite construction microballoon in order to make; Can utilize cationic polyelectrolyte as molecule glue; It is adsorbed in diacetylene vesica surface, continues absorption diacetylene vesica then on the cationic polyelectrolyte surface, so repeatedly several times.
In above-mentioned polystyrene/diacetylene vesica complex microsphere, the diacetylene molecule is owing to its unique conjugated structure, and photopolymerization takes place in the transition that under ultraviolet lighting, can produce delocalized, presents distinctive blueness.When the skeleton that gathers diacetylene when blueness received the interference of environmental stimuli, the conformation of polymer backbone can change, and causes the variation of delocalized electron state, causes its absworption peak blue shift, makes the color transition that gathers diacetylene for red.
Two of the object of the invention provides a kind of polystyrene with metachrosis and bio-sensing effect/gather diacetylene vesica complex microsphere.
Polystyrene provided by the present invention/gather diacetylene vesica complex microsphere obtains above-mentioned polystyrene/diacetylene vesica complex microsphere behind UV-irradiation; Wherein, the optimum condition of said UV-irradiation is following: irradiation is 1-2 minute under the uviol lamp of 254nm, 60w.
Polystyrene diacetylene composite construction microspheres solution issues third contact of a total solar or lunar eclipse polymerization at ultraviolet lighting, forms blue monodispersity solution (being that polystyrene gathers the diacetylene complex microsphere), and this solution can be to temperature, pH, and solvent, light, biomolecule etc. are discerned detection.Said polystyrene gathers diacetylene composite construction microballoon receive above-mentioned external condition and stimulate after, change color takes place, under the exciting of 514-515nm wavelength, can inspire red fluorescence at the 527nm place.
Three of the object of the invention provides a kind of biology sensor.
Biology sensor provided by the present invention, it comprises: polystyrene/the gather conjugate of diacetylene complex microsphere and biomolecule; Said biomolecule has the specific recognition effect to determinand.
The pairing of said determinand and biomolecule is like antigen-antibody, glycoprotein etc.
Biology sensor to detect influenza virus is an example, and the preparation method of above-mentioned polystyrene/the gather conjugate of diacetylene complex microsphere and Antibody of Influenza is elaborated.
Antibody of Influenza is coupled to the surface that said polystyrene gathers diacetylene (PSPDA) composite construction microballoon, and formation antibody polystyrene/gather diacetylene multi-layer compound structure microballoon detects the influenza virus in the solution (like H5N1 virus) effectively.
Said coupling method is following: the diacetylene that gathers on polystyrene spheres surface passes through cross-linking reaction; Can produce the active ester group on the surface; This group can be had an effect with the amido of Antibody of Influenza, thereby makes antibody be connected to gathering on the diacetylene vesica of polystyrene microsphere.This reaction needed is operated in super-clean bench, and in the process that connects antibody, the low temperature that needs maintenance to be no more than 4 ℃ is operated.Because PSPDA composite construction microballoon can centrifugalize under certain rotating speed, has avoided separation method---the dialysis of conventional method, thereby has saved time, and helps keeping the activity of antibody again, the centrifugal antibody that does not connect of removing.In the coupling after the antibody, use sealer (like BSA etc.) to carry out the sealing of avtive spot, same centrifugally remove unnecessary sealer.The above-mentioned active microballoon that is connected with antibody probe that obtains can carry out the response of rapid sensitive to antigen corresponding in the solution.
The preparation of composite construction microballoon provided by the invention is to utilize polyelectrolyte as molecule glue, and the acetylene vesica of amassing wealth by heavy taxation that will have the sensing effect is fixed in the surface of polystyrene microsphere, forms the composite construction with bio-sensing detection effect.This composite construction has stable for the sensor of other forms of diacetylene, is easy to preserve easy separation, the advantage of strong interference immunity; For immunologic detection methods such as ELISA, have simple, quick again; Naked eyes are visible, lower-price characteristic, meanwhile; This composite construction microballoon also in microreactor, is used as sensing, various aspects such as pharmaceutical carrier.
Description of drawings
Fig. 1 prepares the schematic flow sheet of composite construction microballoon for the present invention.
Fig. 2 is the stereoscan photograph of the diacetylene vesica of embodiment 1 preparation.
Fig. 3 is the stereoscan photograph of the PSPCDA composite construction microballoon of polystyrene microsphere and embodiment 2 preparations.
The Zeta potential of Fig. 4 embodiment 2 preparation PSPDA systems is with the change curve of the absorption number of plies.
Fig. 5 is imaging and the local energy curve distribution of PSPDA composite construction microballoon under fluorescence microscope of embodiment 2 preparations.
Fig. 6 adsorbs the color picture after the different numbers of plies are gathered the diacetylene vesica for PS microballoon among the embodiment 2; And the uv absorption peak value of PSPDA under 640nm is with the change curve of the PDA vesica absorption number of plies.
Fig. 7 adsorbs the stereoscan photograph after the different numbers of plies are gathered the diacetylene vesica for PS microballoon among the embodiment 2.
Fig. 8 is the stereoscan photograph of the PSPDA composite construction microballoon of embodiment 3 preparations.
Fig. 9 is the transmission electron microscope and the stereoscan photograph of the PSPDA composite construction microballoon of embodiment 4 preparations.
Figure 10 is fluorescence intensity block diagram and the fluoroscopic image comparison diagram that the antibody-PSPDA composite construction microballoon of embodiment 5 preparations detects the variable concentrations H5N1 virus.
The specific embodiment
The present invention will be described through specific embodiment below, but the present invention is not limited thereto.
Experimental technique among the following embodiment like no specified otherwise, is conventional method.Used test material among the following embodiment like no specified otherwise, is commercial sources and obtains.
The preparation of embodiment 1, diacetylene vesica solution
One, the purification of diacetylene (PCDA)
The powder of diacetylene (PCDA) molecule is dissolved in the chloroformic solution, sees through filter paper and remove polymer, at ELELA
N-1100Vacuum is revolved and is steamed in the appearance 45 ℃ and chloroform solvent is all revolved dried, obtains the white solid powder.Lucifuge, low-temperature vacuum drying is preserved for use.
Two, the preparation of diacetylene vesica solution
Diacetylene (PCDA) monomer powders of purifying is taken by weighing 1.87mg join in the 10mL phosphate buffer solution (pH=7.4), utilize the ultrasonic cell pulverization appearance of SCIENIZ JY92-II type, ultrasonic 5min, the clear solution that obtains clarifying, the concentration of PCDA is 0.5mM.The soln using tinfoil lucifuge that makes is spent the night under 4 ℃.Utilizing the aperture next day is to store for future use after 0.2 μ m-0.5 μ m film filters.This preparation method is with respect to the preparation of traditional diacetylene vesica solution; Have simple to operate, stable, characteristics such as good reproducibility; On the diacetylene formulations prepared from solutions, be beyond example, this is that the preparation of back core-shell material and the stability of keeping this material are laid a good foundation.Its electromicroscopic photograph is as shown in Figure 2.
(the polystyrene microsphere finishing of purchasing has sulfonic group electronegative to eccentric cleaning under the centrifugal force of 4000g with polystyrene microsphere for use (the about 1 μ m of particle diameter); Zeta potential is-28.6mV), get the 3.4mL mass fraction and be 0.134% polystyrene microsphere solution (nearly 10
10Individual), the pH value of HCl standard liquid regulator solution of using 1M adds polymine (PEI) (Mw 50000) solution (concentration that contains NaCl is 0.1M) of 1mL, 1mg/mL as between the 3-3.2, leaves standstill hatching 20min.At centrifugal force is under the 4000g-8000g, centrifugally removes unnecessary PEI, adds isopyknic deionized water.Repeated centrifugation, washing three times, this moment PSPEI microspheres solution positively charged, Zeta potential is 63.3mV.The pH value of regulator solution is 3.5, adds kayexalate (Mw 70000) the PSS solution (concentration that contains NaCl is 0.1M) of 1mg/mL, leaves standstill 20min; At centrifugal force is under the 4000g-8000g; Centrifugally remove unnecessary PSS, be the concentration of maintenance system, add isopyknic deionized water.At said PSS surface adsorption PEI, obtain PSPE again
3The microspheres solution of (promptly having wrapped three strata electrolyte), this solution positively charged, Zeta potential are 60.4mV.
The PCDA vesica solution of getting 500 μ L 0.5mM embodiment, 1 preparation joins PSPE
3In the microspheres solution, standing adsorption 30min is under the situation of 4000g at centrifugal force, centrifugally removes the PCDA vesica that end absorption is got on for three times.Repeat to add the operation 2 times of PEI, PCDA vesica, obtain PSPE
3(PEIPCDA)
3Composite construction solution, Zeta potential is-19.The composite construction microspheres solution that obtains is at 254nm at wavelength, and power is that the uviol lamp of 60w shines 2min down, promptly obtains blue solution, and solution is the PSPDA composite construction microspheres solution of (PDA is for gathering diacetylene).Along with the increase of PDA vesica adsorbance, solution increases at the absorbance of 640nm thereupon, i.e. absorption to red ripple increases, and the blue intensities of solution increases.Like Fig. 3 is the ESEM picture of PSPDA composite construction microballoon and the ESEM picture of polystyrene microsphere.Fig. 4 is the change curve of the Zeta potential of PSPDA system with the absorption number of plies.The PSPDA composite construction microballoon that embodiment 2 obtains is receiving thermostimulation (being heated to 40 ℃); After change color takes place; Under the exciting of 514nm wavelength, can inspire red fluorescence at the 527nm place, under fluorescence microscope, demonstrate red aperture; As shown in Figure 5, the Energy distribution on aperture surface representes that the PDA vesica is evenly distributed in the surface of PS microballoon simultaneously.
Fig. 6 gathers color picture and the uv absorption peak value of PSPDA under 640nm behind the diacetylene vesica with the change curve of the PDA vesica absorption number of plies for the PS microballoon adsorbs the different numbers of plies.Can know that by Fig. 6 along with the increase that gathers diacetylene vesica absorption quantity, it is blue that the variation of solution colour becomes gradually, and increase gradually in the absorption value of 640nm.
Fig. 7 adsorbs the ESEM picture (a, c are the picture behind absorption one deck PDA vesica, and b, d are the picture behind three layers of PDA vesica of absorption) after the different numbers of plies are gathered the diacetylene vesica for the PS microballoon.
(the polystyrene microsphere finishing of purchasing has sulfonic group electronegative to eccentric cleaning under the centrifugal force of 4000g with polystyrene microsphere for use (the about 1 μ m of particle diameter); Zeta potential is-28.6mV), get the 3.4mL mass fraction and be 0.134% polystyrene microsphere solution (nearly 10
10Individual), using the pH value of the HCl standard liquid regulator solution of 1M is 7.5, adds PAH hydrochloride (PAH) (Mw 70000) solution (concentration that contains NaCl is 0.3M) of 1mL1mg/ml, leaves standstill hatching 20min.At centrifugal force is under the 4000g-8000g, centrifugally removes unnecessary PAH, adds isopyknic deionized water.Triplicate, this moment PSPAH microspheres solution positively charged, Zeta potential is about 15.2.The pH value of regulator solution is 3.5, adds the polystyrene sodium sulphate PAA solution (concentration that contains NaCl is 0.3M) of 1mg/ml, leaves standstill 20min; At centrifugal force is under the 4000g-8000g; Centrifugally remove unnecessary PAA, be the concentration of maintenance system, add isopyknic deionized water.Repeat aforesaid operations several times, obtain PSPE
3The microspheres solution of (promptly having wrapped three strata electrolyte), this solution positively charged, Zeta potential are 16.8.
The PCDA vesica solution of getting 500 μ L 0.5mM embodiment, 1 preparation joins PSPE
3In the microspheres solution, standing adsorption 30min is under the situation of 4000g at centrifugal force, removes the PCDA vesica that not absorption is got on centrifugal three times.Repeat to add the operation 2 times of PAH, PCDA vesica, obtain PSPE3 (PAHPCDA)
3Microspheres solution, Zeta potential is-21.2.Is at 254nm with microspheres solution at wavelength, and power is that the uviol lamp of 60w shines 2min down, promptly obtains blue solution, and solution is the PSPDA composite construction microspheres solution of (PDA is for gathering diacetylene).Like Fig. 8 is the picture of its ESEM.
With the step among the embodiment 3, but the PS polystyrene microsphere is of a size of about 100nm.Fig. 9 is its transmission electron microscope photo and stereoscan photograph.
The application of the PSPDA composite construction microballoon of embodiment 5, embodiment 2 preparations
1, the preparation of antibody-PSPDA multi-layer compound structure
Get the PSPDA composite construction microspheres solution (blueness) that 1mL embodiment 2 obtains, adding 400 μ L molar concentrations respectively is the NHS of 1mM, EDC solution (solvent is a phosphate buffer solution, and the pH value of solution value is 7.2), and normal temperature activation 20min makes the surface produce the active ester group.At centrifugal force is under the 8000g, centrifugal three times, remove Puiescent, inoperative NHS in the supernatant, and EDC adds isopyknic phosphate buffer solution (pH=7.2) dissolving with lower sediment.The PSPDA microspheres solution that embodiment 2 obtains is easy to centrifugal under the centrifugal force of 4000g, and the vesica solution with respect to PDA has segregative advantage.
In 4 degrees centigrade ice-water bath, add 200 μ L, dilute the antibody (test of ELISA antibody titer is 1: 5000, and experiment shows that optimum detection concentration is 3000 times of dilutions) of 3000 times H5N1 avian influenza virus, in shaking table, hatch 3h.Make the active ester group on PSPDA surface, react, gather on the diacetylene (PDA) thereby antibody is connected to the amido of antibody.Hatching is removed unnecessary antibody with mixed solution eccentric cleaning under the centrifugal force of 4000g after finishing.With respect to traditional dialysis process, so not only saved the time of response analysis, practice thrift experimental cost, and kept the activity of antibody.For fear of specific adsorption, after the antibody, the BSA phosphate buffer solution that adds 400 μ L150 μ g/mL seals avtive spot in the coupling, subsequently, centrifugally under the centrifugal force of 4000g removes unnecessary BSA.
2, the detection of H5N1 virus
Get the solution of above-mentioned binding antibody, 7 groups of parallel laboratory tests are set.In 7 groups of parallel laboratory tests, add H5N1 virus solution respectively in reaction solution, thereby make the final concentration of H5N1 virus in reaction system be respectively 0ng/mL, 0.1ng/mL, 1ng/mL, 5ng/mL, 10ng/mL, 50ng/mL and 100ng/mL.Mix the back and in 36 ℃ insulating box, react 30min.Utilize laser confocal microscope to carry out check and analysis, obtain detection and be limited to 1ng/ml.Fluorescence intensity block diagram and fluoroscopic image comparison diagram that detection obtains are seen Figure 10.Can know that by Figure 10 along with the increase that adds the H5N1 virus amount, the red aperture in the fluoroscopic image simultaneously, can find out that through the fluorescence intensity block diagram fluorescence intensity strengthens gradually by variable gradually secretly.
Claims (11)
1. a method for preparing polystyrene/diacetylene vesica complex microsphere comprises the steps:
1) preparation diacetylene vesica solution;
2) adopt the static self-assembly method at charged polystyrene microsphere surface deposition polyelectrolyte layer, obtain coating the polystyrene microsphere of polyelectrolyte layer; Wherein, said polyelectrolyte layer is at least one deck, and outermost polyelectrolyte is positively charged;
3) in step 2) the diacetylene vesica of the polystyrene microsphere surface adsorption step 1) preparation of the coating polyelectrolyte layer that obtains, obtain said polystyrene/diacetylene vesica complex microsphere.
2. method according to claim 1 is characterized in that: said method also comprises the steps: the surperficial absorption earlier of the polystyrene/diacetylene vesica complex microsphere that obtains in step 3) cationic polyelectrolyte, and then absorption diacetylene vesica; Repeat above-mentioned steps 0-2 time, preferably repeat 1 time.
3. method according to claim 1 and 2 is characterized in that: in the step 1), the method for preparing diacetylene vesica solution is following: the diacetylene monomer powders is added in the phosphate buffer solution, and ultrasonic being uniformly dispersed obtains diacetylene solution; Said diacetylene solution was placed 5-12 hour 0-4 ℃ of following lucifuge, and using the aperture then is that the miillpore filter of 0.2 μ m-0.5 μ m filters, and promptly gets said diacetylene vesica solution.
4. according to each described method among the claim 1-3, it is characterized in that: step 2) in, the particle diameter of said polystyrene microsphere is 100nm-1 μ m, its positively charged or negative electrical charge in surface;
Step 2) polyelectrolyte layer described in is two-layer at least, and it is the sequence layer that is made up of cationic polyelectrolyte and anionic polyelectrolyte alternating deposit, and said polyelectrolyte layer is preferably the 3-6 layer; Wherein, said cationic polyelectrolyte is polymine or PAH hydrochloride, and said anionic polyelectrolyte is kayexalate, polystyrene sodium sulphate or polyacrylic acid.
5. according to each described method among the claim 1-4; It is characterized in that: step 2) in; Said polystyrene microsphere is electronegative polystyrene microsphere, and the preparation method of the polystyrene microsphere of said coating polyelectrolyte layer comprises the steps: in said electronegative polystyrene microsphere solution, to add cationic polyelectrolyte, leaves standstill hatching 15-20min; Centrifugally remove unnecessary polyelectrolyte, obtain coating the polystyrene microsphere solution of one deck polyelectrolyte layer.
6. method according to claim 5; It is characterized in that: the preparation method of the polystyrene microsphere of said coating polyelectrolyte layer also comprises the steps: in the polystyrene microsphere solution of said coating one deck polyelectrolyte layer, to add anionic polyelectrolyte; Leave standstill 15-30min, centrifugally remove unnecessary anionic polyelectrolyte, then add cationic polyelectrolyte again; Leave standstill 15-30min; Centrifugally remove unnecessary cationic polyelectrolyte, repeat above-mentioned steps 0-4 time, obtain coating the polystyrene microsphere of multi-layer polyelectrolyte layer.
7. the polystyrene that each described method prepares among the claim 1-6/diacetylene vesica complex microsphere.
8. a polystyrene/gather diacetylene vesica complex microsphere is that the described polystyrene of claim 7/diacetylene vesica complex microsphere obtains behind UV-irradiation; Wherein, the optimum condition of said UV-irradiation is following: irradiation is 1-2 minute under the uviol lamp of 254nm, 60w.
9. the application of the described polystyrene of claim 8/gather diacetylene vesica complex microsphere in preparation biology sensor or pharmaceutical carrier.
10. biology sensor, it comprises: the described polystyrene of claim 8/the gather conjugate of diacetylene vesica complex microsphere and biomolecule; Said biomolecule has the specific recognition effect to determinand.
11. biology sensor according to claim 10 is characterized in that: said determinand is the H5N1 influenza virus, and said biomolecule is the H5N1 Antibody of Influenza; The conjugate of said polystyrene/gather diacetylene vesica complex microsphere and H5N1 Antibody of Influenza prepares according to following method: what make said polystyrene/gather diacetylene vesica complex microsphere surface gathers diacetylene through cross-linking reaction, produces the active ester group on the surface; The amido of said ester group and H5N1 Antibody of Influenza is reacted, obtain said conjugate.
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| CN107245196A (en) * | 2017-06-22 | 2017-10-13 | 成都爱兴生物科技有限公司 | Multilayer assembles the preparation method of micron size polystyrene magnetic composite microsphere |
| CN113106444A (en) * | 2021-03-22 | 2021-07-13 | 江苏万力机械股份有限公司 | Automobile crankshaft surface flow mark removing treatment method |
| CN114099439A (en) * | 2021-11-19 | 2022-03-01 | 四川大学 | Preparation method of polybutylene vesicle with excellent performance |
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| CN101701960A (en) * | 2009-11-18 | 2010-05-05 | 中国科学院化学研究所 | Compound for detecting microcystin and application thereof |
| KR20120081762A (en) * | 2011-01-12 | 2012-07-20 | 경희대학교 산학협력단 | Polydiacetylene based biosenser |
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| CN114099439A (en) * | 2021-11-19 | 2022-03-01 | 四川大学 | Preparation method of polybutylene vesicle with excellent performance |
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