CN1963524A - A molecule switching type miniflow control chip - Google Patents
A molecule switching type miniflow control chip Download PDFInfo
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- CN1963524A CN1963524A CN 200610118101 CN200610118101A CN1963524A CN 1963524 A CN1963524 A CN 1963524A CN 200610118101 CN200610118101 CN 200610118101 CN 200610118101 A CN200610118101 A CN 200610118101A CN 1963524 A CN1963524 A CN 1963524A
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Abstract
This invention provides one molecule switch micro flow control chip in chemical field, which comprises chip base, tank channel, sample in hole, sample out hole on both ends of tank channel, wherein, tank inner surface is coated with metal film and decorated with C8-C24 hydrocarbon linkage to selectively absorb positive or negative large molecule through outside level operation control chip.
Description
Technical field
The present invention relates to Separation of Proteins in the chemical field, facture of microchip technology, supermolecule synthetic technology and self-assembling technique.Particularly, the invention provides a kind of molecule switching type miniflow control chip that can be used for big molecular separation.
Background technology
Molecular switch be on the structure systematism the chemical system with " ON/OFF " function, it is in the existing bigger progress of the application of aspects such as optics, biology, medical science, and also is important foundation for the development of molecular computer.According to the function and the purposes of molecular switch, can it be divided into magnetic molecule switch, fluorescence molecule switch, logic molecular switch, characteristic functions molecular switch etc. (Wu Biyao, Zhang Daohong, New Chemical Materials, the 29th the volume o. 11th, November calendar year 2001,9-13).
(self-assembled monolayer is the materialization effect to take place and a kind of Thermodynamically stable of spontaneous formation, individual layer (or multilayer) molecular film of queueing discipline between structure membrane molecule and base material SAM) to self assembled monolayer.Adopt the superthin layer system of this technology preparation to mainly contain two classes: a class is the individual layer that sulfhydryl compound forms in gold, silver, copper, platinum surface adsorption; One class is the individual layer that forms by Silanization reaction on surfaces such as silicon, glass and metal oxides.Wherein the former research is more extensive, the formation of SAM based on the aligning of strong chemical bonding of sulfydryl and base material and polymethylene chain (Dong Xiandui, Lu Juntao look into full property, galvanochemistry, the 1st the 3rd phase of volume, August nineteen ninety-five, 248-258).
Have the low-density self assembled monolayer that the thiol molecule of the long-chain of special end group forms and under the driving of external condition, to present " ON/OFF " conversion, can be used as molecular switch.The result of study of J.Lahhann etc. shows, self assembly is electronegative owing to its end group carboxyl at the low-density 16 mercapto acid molecules of gold surface, voltage on the golden face that can apply by change, make it present the state of bending (positive voltage) or upright (negative voltage), thereby make whole surface be the state of hydrophobic or hydrophilic (and electronegative), i.e. " ON/OFF " state (J.Lahann, S.Mitragotri, T.N.Tran, H.Kaido, J.Sundaram, I.S.Choi, S.Hoffer, G.A.Somorjai and R.Langer, Science, 2003,299,371-374.).
And the method for controlling 16 mercapto acid molecule density comprises: have the 16 mercapto acid derivatives of bulky group or inclusion compound (the Ying Liu of acid of 16 mercaptos and macrocyclic compound (cyclodextrin, calixarenes etc.) on the synthetic carboxyl, Li Mu, Baohong Liu, Song Zhang, Pengyuan Yang and Jilie Kong, Chem.Commun., 2004,1194-1195.), after self-assembled film forms, manage to remove bulky group or macrocyclic compound again, then can obtain low-density 16 mercaptos acid unimolecular layer.Use similar method, also the end group that can make is the low-density unimolecular layer of the amino long chain molecule that waits other functional group.The surface that the hydrophilic and hydrophobic matter that obtains like this can be controlled can be used for separating the big molecules such as protein of the different isoelectric points of tool.Even can make the chirality switch on this basis.
Micro-fluidic chip is to become circuit the same function element image sets such as microchannel, little reservoir, microelectrode by Micrometer-Nanometer Processing Technology, makes them be integrated in micro-total analysis system on the chip material.Its manufacturing materials is any organic material such as PMMA, PDMS polymkeric substance or quartz, glass.Manufacture craft comprises technology such as hot pressing, wet etching, photoetching (Luo Yi, Lou Zhifeng, Chu Denan, Liu Chong, Wang Liding, nanometer technology and precision engineering, the 2nd the 1st phase of volume, in March, 2004,20-23.)。
Summary of the invention
The purpose of this invention is to provide a kind of macromolecular molecule switching type miniflow control chip that can be used for separating.
Another object of the present invention provides the preparation method of said chip.
The invention provides a kind of molecule switching type miniflow control chip, comprise tabular chip matrix, elongated shape conduit, injection port, outlet, injection port and outlet lay respectively at the two ends of conduit, the conduit inside surface is coated with golden film, and be modified with chain length and be 8-24 carbon, an end has the straight-chain paraffin molecule of sulfydryl, the distribution density of this alkane molecule is 0.99-2.08nmol/cm on the golden film
2
Among the present invention, described chain length is that the end group of other end 8-24 carbon, that an end has a straight-chain paraffin molecule of sulfydryl is carboxyl or amino.
Among the present invention, described chip material can be any suitable material.For example, polymethylmethacrylate, dimethylsiloxane polymer, quartz or glass etc.
Among the present invention, the size visual exam condition of chip and each several part thereof and requirement of experiment and decide, for example, the conduit size can be that length is 3～8cm, diameter 80～200 μ m, the degree of depth 40～100 μ m; The degree of depth of injection port and outlet is identical with conduit, and diameter is 300 μ m-2cm.
Among the present invention, the chain length of described hydrocarbon chain molecule can be C
16-C
18
The used decorating molecule of chip of the present invention is that (preparation is during supermolecule for the supermolecule of long-chain mercapto molecule and cyclodextrin, use alpha-cyclodextrin, beta-schardinger dextrin-, gamma-cyclodextrin all can, because varying in size of three kinds of cyclodextrin, the density difference of the thiol molecule layer of its control, but all has the switch character that impressed voltage is controlled), or with the long-chain mercapto molecule of special bulky group, with self-assembly method chip conduit surface is modified, removed the cyclodextrin or the bulky group of control of density then with special solvent wash-out or special reaction such as hydrolysis reaction etc.
On the other hand, the invention provides the preparation method of said chip, this method may further comprise the steps:
(1) on matrix, makes conduit, injection port and outlet;
(2) gold-plated at the conduit inside surface, golden film thickness is 350-450nm;
(3) will be combined with earlier cyclodextrin or calixarenes and the band sulfydryl C
8-C
24The hydrocarbon chain molecule combines with golden film, in conjunction with after remove cyclodextrin or calixarenes again, clean the standby described chip that promptly obtains.
Among the present invention, the C of band sulfydryl in the above-mentioned steps (3)
8-C
24The hydrocarbon chain molecule can inject the ethylenediamine solution of 0.05～5mol/L again with the carboxyl on the carboxyl activator activation hydrocarbon chain molecule then with after golden film combines, and cleans standbyly at last, promptly obtains the C by band hydrosulfamine group
8-C
24The chip of hydrocarbon chain molecular modification conduit.
Among the present invention, carboxyl activator can be a carbodiimide class carboxyl activator in the step (3).For example, ethyl-(3-dimethyl propyl) carbodiimide hydrochloride (EDC) and N-alkyl sulfosuccinimide (NHS) or ethyl-(3-dimethyl propyl) carbodiimide hydrochloride (EDAC) for example.
Conduit method for making among the present invention can be taked pressure sintering, laser ablation, monomer in-situ polymerization etc.The method of chip pressing is a pressure sintering.
The golden face method for making in conduit surface can be used ion sputtering method, metal evaporation method etc. among the present invention.
Among the present invention, sample introduction goes out the sample available pump or other air pressure drives.
Among the present invention, step (3) can be finished according to the following steps: the at first synthetic supermolecule solution that is used to modify, with α-, β-or gamma-cyclodextrin and long-chain mercapto molecule (8-24 carbon) by synthesizing greater than 2: 1 mol ratio, the long-chain mercapto molecule is added in the saturated rings dextrin saturated aqueous solution (or oversaturated suspension), heated and stirred, 35～60 ℃ of temperature of reaction.48～96 hours reaction time.Products therefrom is got the conduit that solution partly injects chip after filtering and is assembled, 4～25 ℃ of 8-24 hour (24 hours best) of assembling down.Remove cyclodextrin molecular with the appropriate solvent wash-out then, wash-out soak time 5～30 minutes is fully washed with deionized water again.If with long-chain mercapto molecular modification, then need after self assembly, to remove substantially actively group with suitable reactions such as hydrolysis reaction with special bulky group.Can obtain the chip of low-density long-chain mercapto molecular modification.
If will modify the mercapto acid molecule of terminal band carboxyl, modification gets final product as stated above.If will modify the amino hydrosulfamine molecule of terminal band, then need on the basis of modifying the mercapto acid molecule, further to modify.Concrete grammar is: earlier activate on the 16 mercaptos acid unimolecular layer carboxyl 30-60 minute with carboxyl activator.Carboxyl activator can be the carbodiimide class, for example ethyl-(3-dimethyl propyl) carbodiimide hydrochloride (EDC) and N-alkyl sulfosuccinimide (NHS) or ethyl-(3-dimethyl propyl) carbodiimide hydrochloride (EDAC).Use deionized water rinsing then, the ethylenediamine solution of the 0.05～5mol/L that reinjects, under saturated humidity, kept at room temperature overnight.It is standby that deionized water fully cleans the back.
During chip application of the present invention, working electrode can be linked to each other with outlet, injection port links to each other with contrast electrode with comparison electrode, add 0.1～0.4 volt of positive voltage or negative voltage-0.1～-0.4 volt on the golden film on conduit surface, the sample solution of tape label is finished the separation of albumen along conduit to outlet by injection port.
Chip of the present invention can be used for separating the big molecule of the different electric charges of band of concentration range 0.02～0.2mg/mL.For example, with the big molecule of this chip: under a certain impressed voltage at the different electric charges of separable band under the impressed voltage condition, the big molecule of a kind of electric charge of chip selective adsorption band, and under another kind of impressed voltage the big molecule of the another kind of electric charge of adsorption band optionally.
For example, separate the different protein of isoelectric point with this chip.
The phosphate buffer solution of implantation concentration 5mmol/L ~ 100mmol/L in chip at first.The pH value will satisfy makes two kinds of different electric charges of separated protein band, and promptly selected pH value will be between the isoelectric point of separated albumen, and guarantees to make the end group that is modified at the molecule on the chip conduit inside surface can dissociate charged.If be used for the chip that mercapto acid is modified, buffer solution pH6.8～11.5; If be used for the chip that hydrosulfamine is modified, buffer solution pH5.1～7.2; If be used for the chip of mercapto acid, hydrosulfamine mixing modification, buffer solution pH5.1～11.5, the gold-plated zone of chip egative film and the working electrode of electrochemical workstation or other potentiometer are linked to each other, in the injection port of chip, insert contrast electrode and to electrode, apply voltage (malleation or negative pressure by electrochemical workstation or other potentiometer then, according to the protein of required absorption with electric charge decide, and must satisfy selected voltage in the burning voltage scope of self assembly layer and protein.Malleation 0.1～0.4V; Negative pressure-0.1～-0.4V).After waiting that the voltage of executing is stablized certain hour, one side adds 1～20uL mixed protein solution in chip (the pH value is identical with buffering solution; Concentration 0.02～0.2mg/mL), with pump slowly extract unnecessary buffer solution on one side, must guarantee that the buffer solution of extraction and the mixed protein liquor capacity of adding equate.Continued to apply voltage 10 ~ 60 minutes.Applying under the voltage condition then, in chip, replenish buffer solution on one side, on one side extract the solution that chip dashes slowly out, guarantee that the volume of solution of extracting out and the mixed protein solution that injects before is equal, can obtain containing not the solution of the protein that is adsorbed by chip with pump.After this with executing voltage reversal, and kept 10～60 minutes, the protein that is adsorbed is released.
This detachment process not only is suitable for for protein, and also is suitable for the big molecule of different electric charges or the separation of nano material for other.
The invention provides a kind of molecule switching type miniflow control chip, comprise chip matrix, conduit, injection port, outlet, injection port and outlet lay respectively at the two ends of conduit, and the conduit inside surface is coated with golden film, and are modified with the C of band sulfydryl
8-C
24Hydrocarbon chain.This chip accuracy height uses simple and easyly, can be used for separating the big molecule of the different electric charges of band of concentration range 0.02～0.2mg/mL.
Description of drawings
Fig. 1 is macromolecular principle schematic for molecule switching type miniflow control chip separates.
Fig. 2 is embodiment 3 fluorogram as a result.Wherein, straight line is represented to add-fluorogram of the mixed protein solution that flows out from chip during 0.3V voltage; Dotted line is voltage transition is represented the mixed protein solution that flows out behind the 0.3V from chip a fluorogram; Green (near axis of ordinates) be the fluorogram of antibiotin avidin; Red (away from axis of ordinates) be the fluorogram of strepto-biotin streptavidin.
Fig. 3 is embodiment 4 fluorogram as a result.Wherein, straight line is represented to add-fluorogram of the mixed protein solution that flows out from chip during 0.3V voltage; Dotted line is voltage transition is represented the mixed protein solution that flows out behind the 0.3V from chip a fluorogram; Green (near axis of ordinates) be the fluorogram of antibiotin avidin; Red (away from axis of ordinates) be the fluorescence of strepto-biotin streptavidin.
Fig. 4 is the fluorogram of the mixed protein solution that adds among the embodiment 5-flow out behind the 0.3V voltage from chip.Green (near axis of ordinates) be the fluorogram of antibiotin avidin; Red (away from axis of ordinates) be the fluorescence of strepto-biotin streptavidin.
Fig. 5 is for adding the fluorogram of the mixed protein solution that flows out behind the 0.3V voltage from chip among the embodiment 5.Green (near axis of ordinates) be the fluorogram of antibiotin avidin; Red (away from axis of ordinates) be the fluorescence of strepto-biotin streptavidin.
Fig. 6 shows synoptic diagram for the structure of molecule switching type miniflow control chip.Wherein, 1 is matrix substrate, and 2 is injection port, and 3 is outlet, and 4 is conduit, and 5 is working electrode, and 6 is to electrode, and 7 is contrast electrode.
Embodiment
Embodiment 1
Make a flat recess road in the chip egative film central authorities that with the PDMS polymkeric substance are material with wet etching method, channel lengths 8cm, diameter 80 μ m, the degree of depth 40 μ m.Gold-plated with methods such as metal evaporations then at conduit and egative film one end.Cover last slice bored injection port and outlet after, cover on the egative film with pressure sintering, chip is shaped substantially.Then paste or the weld metal lead, link to each other in order to electrode with making alive equipment (as electrochemical workstation etc.) in the gold-plated zone of egative film.At sticking plastic tube of injection port and sample outlet position or glass tube, as sample inlet pool with go out the sample pipe.It is thinner wherein to go out sample pipe end, is convenient to be connected with pump.
The chip that makes also need be modified with supermolecule in conduit.The at first synthetic supermolecule solution that is used to modify, synthesize by 2: 1 mol ratio with alpha-cyclodextrin and long-chain mercapto molecule (8 carbon), the long-chain mercapto molecule is added in the saturated rings dextrin saturated aqueous solution heated and stirred, 60 ℃ of temperature of reaction, 48 hours reaction time.Products therefrom is got the conduit that solution partly injects chip after filtering and is assembled, 25 ℃ of assemblings 24 hours down.Remove cyclodextrin molecular with the dimethyl sulfoxide wash-out then, wash-out soak time 30 minutes is fully washed with deionized water again.If with long-chain mercapto molecular modification, then need after self assembly, to remove substantially actively group with suitable reactions such as hydrolysis reaction with special bulky group.Can obtain the chip of low-density long-chain mercapto molecular modification.
Make a flat recess road in the chip egative film central authorities that with the quartz are material with pressure sintering, channel lengths 3cm, diameter 200 μ m, the degree of depth 100 μ m.Use the ion sputtering method gold-plated then at conduit and egative film one end.Cover last slice bored injection port and outlet after, cover on the egative film with pressure sintering, chip is shaped substantially.Then directly link to each other standby with making alive equipment.
The chip that makes also need be modified with supermolecule in conduit.The at first synthetic supermolecule solution that is used to modify, synthesize by 4: 1 mol ratio with calixarenes and long-chain mercapto molecule (24 carbon), the long-chain mercapto molecule is added in the supersaturation cyclodextrin saturated aqueous solution heated and stirred, 35 ℃ of temperature of reaction, 96 hours reaction time.Products therefrom is got the conduit that solution partly injects chip after filtering and is assembled, 5 ℃ down assembling spend the night.Remove the calixarenes molecule with the absolute ethyl alcohol wash-out then, wash-out soak time 5 minutes is fully washed with deionized water again.If with long-chain mercapto molecular modification, then need after self assembly, to remove substantially actively group with suitable reactions such as hydrolysis reaction with special bulky group.Can obtain the chip of low-density long-chain mercapto molecular modification.
On the basis of modifying the mercapto acid molecule, further modify subsequently.Earlier activate carboxyl 60 minutes on the 16 mercaptos acid unimolecular layer with carboxyl activator.Carboxyl activator is ethyl-(3-dimethyl propyl) carbodiimide hydrochloride (EDAC).Use deionized water rinsing then, the ethylenediamine solution of the 5mol/L that reinjects, under saturated humidity, kept at room temperature overnight.It is standby that deionized water fully cleans the back, promptly obtains the chip of the amino hydrosulfamine molecular modification of terminal band.
Chip with antibiotin (avidin) and streptavidin (streptavidin) mixed protein solution and the modification of low-density 16 mercaptos acid unimolecular layer is an example.The molecular weight of these two kinds of protein is respectively 67kDa and 52.8kDa, isoelectric point is respectively 10.5 and 5, and use Fluorescein (fluorescein respectively, excitation wavelength 494nm, emission wavelength 518nm) and Texas Red (texas Red, excitation wavelength 595nm, emission wavelength 615nm) carried out fluorescence labeling.
Used chip material is organic material PMMP, and channel lengths is 5cm, diameter 150 μ m, the degree of depth 75 μ m.
Chip is modified used supermolecule solution by beta-schardinger dextrin-(β-CD) made in 48 hours at 40 ℃ of following stirring reactions by 4: 1 mol ratio with 16 mercapto acid (MHA).The time of supermolecule solution self assembly in the chip conduit is 24 hours, 20 ℃ of assembling temperature.When the absolute ethyl alcohol wash-out is removed cyclodextrin, soaked 15 minutes earlier, fully flushing is then fully washed with deionized water again.
During Separation of Proteins, select 10mmol/L for use, the phosphate buffer solution of pH7.4, apply-0.3V voltage 30 minutes absorption avidin protein is (under this pH value condition, self assembled monolayer is electronegative, the avidin positively charged, and streptavidin is electronegative), utilize the effect of pump then, remaining streptavidin solution is gone out slowly with buffer solution.Then change into and applied 0.3V voltage 30 minutes, the avidin protein that is adsorbed is released, and rushes out with buffer solution.The solution of going out for twice can be used fluoroscopic examination (Fig. 2).The proof mixed protein has obtained separation.
Embodiment 4
Chip with antibiotin (avidin) and streptavidin (streptavidin) mixed protein solution and the modification of low-density 16 hydrosulfamine unimolecular layers is an example.The molecular weight of these two kinds of protein is respectively 67kDa and 52.8kDa, isoelectric point is respectively 10.5 and 5, and use Fluorescein (fluorescein respectively, excitation wavelength 494nm, emission wavelength 518nm) and Texas Red (texas Red, excitation wavelength 595nm, emission wavelength 615nm) carried out fluorescence labeling.
Used chip is a flat recess road chip, and the conduit two ends are respectively injection port and outlet, and material is organic material PMMP, and channel lengths is length 5cm, diameter 150 μ m, the degree of depth 75 μ m.
The modification of chip was divided into for two steps, and the first step is the same with the preparation method of 16 mercaptos acid unimolecular layer, at first makes supermolecule solution with the acid of beta-schardinger dextrin-16 mercaptos in 48 hours by 40 ℃ of following stirring reactions of mol ratio of 4: 1.The time of supermolecule solution self assembly in the chip conduit is 24 hours, and the assembling temperature is 20 ℃.When the absolute ethyl alcohol wash-out is removed beta-schardinger dextrin-, soaked 15 minutes earlier, fully flushing is then fully washed with deionized water again.Second step, (EDC concentration is 0.05mol/L with carboxyl activator EDC/NHS earlier, NHS concentration is 0.2mol/L, all be to use 10mmol/L, the phosphate buffer solution of pH7.4 is freshly prepared) carboxyl on the activation 16 mercaptos acid unimolecular layer 60 minutes, use deionized water rinsing then, the ethylenediamine solution of the 1mol/L that reinjects, under saturated humidity, kept at room temperature overnight.After deionized water fully cleans, promptly can be used for Separation of Proteins.
During Separation of Proteins, select 10mmol/L for use, the phosphate buffer solution of pH6.6, apply 0.3V voltage absorption in 30 minutes streptavidin protein (under this pH value condition, the self assembled monolayer positively charged, the avidin positively charged, and streptavidin is electronegative), utilize the effect of pump then, remaining avidin solution is gone out slowly with buffer solution.Then change into and applying-0.3V voltage 30 minutes, the streptavidin protein that is adsorbed is released, and rushes out with buffer solution.The solution of going out for twice can be used fluoroscopic examination (Fig. 3).The proof mixed protein has obtained separation.
Mixing the chip of modifying with antibiotin (avidin) and streptavidin (streptavidin) mixed protein solution and low-density 16 mercaptos acid unimolecular layer, 16 hydrosulfamine unimolecular layers is example.The molecular weight of these two kinds of protein is respectively 67kDa and 52.8kDa, isoelectric point is respectively 10.5 and 5, and use Fluorescein (fluorescein respectively, excitation wavelength 494nm, emission wavelength 518nm) and Texas Red (texas Red, excitation wavelength 595nm, emission wavelength 615nm) carried out fluorescence labeling.
Used chip structure as shown in Figure 1, material is organic material PMMP, channel lengths is length 5cm, diameter 150 μ m, the degree of depth 75 μ m, working electrode links to each other with outlet, injection port links to each other with contrast electrode with comparison electrode.
The modification of chip was divided into for two steps, and the first step is at first used beta-schardinger dextrin-(β-CD) 48 hour make supermolecule solution by 4: 1 mol ratio at 40 ℃ of following stirring reactions with 16 mercapto acid (MHA).The time of supermolecule solution self assembly in the chip conduit is 24 hours, and the assembling temperature is 20 ℃.When the absolute ethyl alcohol wash-out is removed beta-schardinger dextrin-, soaked 15 minutes earlier, fully flushing is then fully washed with deionized water again.Second step, (EDC concentration is 0.05mol/L with carboxyl activator EDC/NHS earlier, NHS concentration is 0.2mol/L, all be to use 10mmol/L, the phosphate buffer solution of pH7.4 is freshly prepared) carboxyl on the activation 16 mercaptos acid unimolecular layer 60 minutes, must use air pressure to control half zone that it rests on the chip conduit when injecting EDC/NHS, make the 16 mercaptos acid unimolecular layer of this part obtain activation.Behind deionized water rinsing, inject the ethylenediamine solution of 1mol/L, under saturated humidity, kept at room temperature overnight.After deionized water fully cleans, promptly can be used for the controlled separation of protein.
During Separation of Proteins, select 10mmol/L for use, the phosphate buffer solution of pH7.0 (under this pH value condition, electronegative, the 16 hydrosulfamine unimolecular layer positively chargeds of 16 mercaptos acid unimolecular layer, the avidin positively charged, and streptavidin is electronegative).
If apply-0.3V voltage 30 minutes earlier to chip, utilize the effect of pump then, go out slowly with the streptavidin solution that buffer solution will not be adsorbed.Then change into and applied 0.3V voltage 30 minutes, the avidin protein that is adsorbed is released, and rushes out with buffer solution.The solution of going out for twice can be used fluoroscopic examination (Fig. 4).
And if elder generation applied 0.3V voltage 30 minutes to chip, utilize the effect of pump then, go out slowly with the avidin solution that buffer solution will not be adsorbed.Then change into and applying-0.3V voltage 30 minutes, the streptavidin protein that is adsorbed is released, and rushes out with buffer solution.The solution of going out for twice can be used fluoroscopic examination (Fig. 5).
The result shows that under different impressed voltages, the release conditions of different proteins is obviously different.Promptly, if earlier chip is applied-voltage of 0.3V, the solution of then going out from chip has stronger fluorescent emission at 615nm, and very weak in the fluorescent emission at 518nm place, illustrate that at this moment what flow out earlier mainly is streptavidin from chip, chip absorption be avidin; If the 0.3V voltage that elder generation applies to chip, the solution of then going out from chip has stronger fluorescent emission at 518nm, and the fluorescent emission at 615nm place is very weak, illustrate that what flow out in the chip at this moment mainly is avidin, and chip absorption is streptavidin.This prove this chip can alive outside control under the protein of the different electric charges of adsorption band optionally.
Claims (10)
1. molecule switching type miniflow control chip, comprise tabular chip matrix, elongated shape conduit, injection port, outlet, injection port and outlet lay respectively at the two ends of conduit, it is characterized in that, the conduit inside surface is coated with golden film, and be modified with chain length and be 8-24 carbon, an end has the straight-chain paraffin molecule of sulfydryl, the distribution density of this alkane molecule is 0.99-2.08 nmol/cm on the golden film
2
2. chip as claimed in claim 1 is characterized in that, described chain length be 8-24 carbon, an end has the straight-chain paraffin molecule of sulfydryl and the end group of the other end is carboxyl or amino.
3. chip as claimed in claim 1 is characterized in that, described chip material is polymethylmethacrylate, dimethylsiloxane polymer, quartz or glass.
4. chip as claimed in claim 1 is characterized in that, channel lengths is 3～8cm, diameter 80～200 μ m, the degree of depth 40～100 μ m; The degree of depth of injection port and outlet is identical with conduit, and diameter is 300 μ m-2cm.
5. chip as claimed in claim 1 is characterized in that, the chain length of described hydrocarbon chain molecule is C
16-C
18
6. chip production method according to claim 1 is characterized in that this method may further comprise the steps:
(1) on matrix, makes conduit, injection port and outlet;
(2) gold-plated at the conduit inside surface, golden film thickness is 350-450nm;
(3) will be combined with earlier cyclodextrin or calixarenes and the band sulfydryl C
8-C
24The hydrocarbon chain molecule combines with golden film, in conjunction with after remove cyclodextrin or calixarenes again, clean the standby chip as claimed in claim 1 that promptly obtains.
7. as chip production method as described in the claim 6, it is characterized in that the C of band sulfydryl in the step (3)
8-C
24The hydrocarbon chain molecule again with the carboxyl on the carboxyl activator activation hydrocarbon chain molecule, injects the ethylenediamine solution of 0.05～5mol/L with after golden film combines then, cleans standbyly at last, promptly obtains the C by band hydrosulfamine group
8-C
24The chip of hydrocarbon chain molecular modification conduit.
8. as chip production method as described in the claim 7, it is characterized in that carboxyl activator is a carbodiimide class carboxyl activator in the step (3).
9. the application of chip according to claim 1 is characterized in that, this chip is used to separate the molecule of the different electric charges of band of concentration range 0.02～0.2mg/mL.
10. the application process of chip according to claim 1, it is characterized in that, working electrode links to each other with outlet, injection port links to each other with contrast electrode with comparison electrode, add positive voltage or negative voltage on the golden film in conduit surface, the sample solution of tape label is finished the separation of albumen along conduit to outlet by injection port.
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| CN101514990A (en) * | 2008-02-21 | 2009-08-26 | 天津市先石光学技术有限公司 | Sensor for sensing contents of components to be measured in human tissue fluid, fluid channel unit and method for measuring contents of components to be measured in human tissue fluid |
| CN101514990B (en) * | 2008-02-21 | 2014-01-29 | 天津先阳科技发展有限公司 | Sensor for sensing contents of components to be measured in human tissue fluid, fluid channel unit and method for measuring contents of components to be measured in human tissue fluid |
| CN102607999A (en) * | 2011-01-19 | 2012-07-25 | 索尼公司 | Method and device for detecting heavy metal ion in water |
| CN109234158A (en) * | 2018-10-08 | 2019-01-18 | 北京京东方技术开发有限公司 | Biochip and its manufacturing method, operating method, biological detection system |
| US11691150B2 (en) | 2018-10-08 | 2023-07-04 | Boe Technology Group Co., Ltd. | Biological chip, manufacturing method thereof, operation method thereof, and biological detection system |
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