CN106117453A - A kind of macromolecule mother solution and the manufacture method of 3D biochip - Google Patents
A kind of macromolecule mother solution and the manufacture method of 3D biochip Download PDFInfo
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Abstract
The invention discloses a kind of macromolecule mother solution and the manufacture method of 3D biochip, mother solution is by obtaining by liquid phase reactor polymerization after OEGMA and methacrylic acid derivative monomer being dissolved in polar solvent.Surface chemical reaction is transferred to carry out in solution by mother solution synthetic method, disposable synthesizes a large amount of macromolecule mother solutions, improve macromolecule terminal derivative productivity and reaction efficiency, it is to avoid the use of the expensive instruments such as glove box, has saved time and human cost.Make biochip after mother solution is modified, shorten the chip manufacturing cycle, improve the stability of chip quality so that chip can be suitably used for large-scale industrial production demand.The inventive method reaction condition is the gentleest, it is easy to accomplish.The inventive method makes the biochip obtained, and has the most anti-non-specific.
Description
Technical field
The present invention relates to a kind of biochip material and the manufacture method of biochip, particularly to a kind of high score primary and secondary
Liquid and the manufacture method of 3D biochip.
Background technology
Polypeptide, antibody, protein drug is developing quickly over nearly 10 years, its outstanding feature is that specificity is high, and response speed is fast,
Few side effects etc., are particularly suitable for the treatment of the relevant disease of tumor.Current antibody screening is mainly also based on the enzyme connection of 384 orifice plates
Immunity Rapid screening techniques, either in flux or speed, all needs to be improved further.Particularly, during current antibody screening
Needing to read data by two anti-fluorescent labelinies, bring extra financial burden to experiment, potential false positive is inevitable.
It is since 2000, with the high flux of Plexera company of the U.S. screening instrument that interacts as representative, the most unmarked,
High flux screening equipment enters the visual field of people.Its representative products PlexeraHT instrument may in 20 minutes rapid screening high
Reach 5000 interaction point positions, and possess unmarked, kinetic character.From it it has been reported that chip from the point of view of, although have
Numerous protein chips are selective, but develop a low price, and the universal protein chip of stable performance still has greatly
Demand and challenge.
Biochip is generally by glass, and a floor height molecule is modified on the surface such as metal, and by albumen, antibody, medicine divides
Son, the biomolecule such as polypeptide is fixed on surface, is then fabricated to micro-fluidic chip, at Surface testing surface institute's fixed substance and egg
In vain, serum, the interaction of the materials such as cell pyrolysis liquid, and then experiment carries out qualitative and quantitative analysis [1-2] to biological components.When
Before, the Main Patterns that the surface of biochip is modified is first at one layer of unimolecular initiators of chip surface self assembly, then at table
Face carries out Polymer Synthesizing, forms two dimension or three dimensional biological film.The macromolecule that this mode synthesizes, because self space steric hindrance is made
With, can be formed on surface and there is three-dimensional molecular brush structure, there is good anti-non-specific and bio-compatibility.Pass through
Regulation lower curtate self assembly initiator density, can control high molecular density and thickness [3-4] that surface is formed to a certain extent.But
It is that this manufacture method operation complexity, generated time is long, and nearly all reaction is all carried out on surface, causes reaction efficiency extremely low
(about 10%), low to raw material availability.Particularly this step of Polymer Synthesizing, because whole chip to be sealed in without strict oxygen bar
Under part, whole processing procedure needs to operate in glove box, greatly hinders industrialization and the standardization of chip production, actual
The chip quality produced also is difficult to ensure that.Therefore, optimize chip synthesis step the most further, improve chip production efficiency,
Improve chip quality control so that chip really can move towards, from laboratory, the work that factory is a new rich challenge.
List of references:
[1]Christopher Lausted, Zhiyuan Hu1 Leroy Hood and Charles T. Campbell;
SPR Imaging for High Throughput, Label-Free Interaction Analysis;
Combinatorial Chemistry & High Throughput Screening, 2009, 12, 741-751
[2] a kind of chip, prepares and just goes, the method application number 201210371255.X of purposes and screening of medicaments
[3] segmented copolymer progress macromolecule based on ATRP technology circulates a notice of 6 phase 36-43 page, 2006 years
[4]Ma H, He,J, Liu X,Gan J, Jin G, Zhou j. Surface Initiated
Polymerization from Substrates of Low Initiator Density and Its Applications
in Biosensors; ACS applied materials & interfaces. 2010:2:3223-40。
Summary of the invention
It is an object of the present invention to provide the synthetic method of a kind of easy macromolecule mother solution for biochip.
A kind of 3D biochip manufacture method being easily achieved of offer is provided.
The technical solution used in the present invention is:
The synthetic method of a kind of macromolecule mother solution, comprises the steps:
1) OEGMA and methacrylic acid derivative monomer are dissolved in polar solvent, remove bubble removing, keep away under protection gas shielded
Light stirs, and obtains monomer mixed solution A, standby；
2) appropriate complex ligand and catalyst transition metal halide are dissolved in the water, obtain solution B；
3) by monomer mixed solution A and solution B mix homogeneously under protection gas shielded, to reactant liquor variable color；
4) in reactant liquor, add weak reductant, be stirring evenly and then adding into initiator initiated polymerization；
5) after reaction completely, by oxygen-containing gas, aoxidize solution, obtain macromolecule mother solution.
As the further improvement of above-mentioned synthetic method, complex ligand is bipyridyl.
As the further improvement of above-mentioned synthetic method, transition metal halide is copper halide.
As the further improvement of above-mentioned synthetic method, weak reductant is selected from ascorbic acid, glucose or stannous octoate.
As the further improvement of above-mentioned synthetic method, initiator is selected from single sulfydryl halogenated thiols, single mercapto-polyglycol
Oligomer, double sulfydryl halogenated thiols, double mercapto-polyglycol oligomer.
As the further improvement of above-mentioned synthetic method, the mol ratio of OEGMA and methacrylic acid derivative monomer be (1:
4)～(4:1).
As the further improvement of above-mentioned synthetic method, the high molecular number-average molecular weight of synthesis is 500,000～5,000,000.
As the further improvement of above-mentioned synthetic method, methacrylic acid derivative monomer is the methacrylic acid of hydroxyl
Derivatives monomer.
The manufacture method of a kind of 3D biochip, including using on modifying agent macromolecule in above-mentioned macromolecule mother solution
Introduce anchoring group, obtain derivatization macromolecule mother solution；Take derivatization macromolecule mother solution afterwards to be laid on carrier, after reaction completely
Clean and remove loose macromolecule, be dried to obtain 3D biochip.
As the further improvement of above-mentioned manufacture method, anchoring group is selected from-COOH ,-NH2,-OH, epoxy radicals ,-OCH3,-
COCNH2, cyano group, alkynyl, azido.
As the further improvement of above-mentioned manufacture method, carrier is for having nanometer gold, nanometer silver, nanometer chromium, three oxidations two
Aluminum or the chip of titanium dioxide surface.
The invention has the beneficial effects as follows:
The macromolecule mother solution synthetic method of the present invention, transfers to surface chemical reaction carry out in solution, and disposable synthesis is a large amount of
Macromolecule mother solution, improves macromolecule terminal derivative productivity and response time, it is to avoid the use of the expensive instruments such as glove box, joint
About time and human cost.
This programme improves chip manufacturing flow process and technique, shorten the chip manufacturing cycle, it is to avoid the costlinesses such as glove box
The use of instrument, has saved time and human cost.Transferring to surface chemical reaction carry out in solution, disposable synthesis is a large amount of
Macromolecule mother solution, improves macromolecule terminal derivative productivity and response time, adds the stability of chip quality so that chip
Can be suitably used for large-scale industrial production demand.
The inventive method reaction condition is the gentleest, it is easy to accomplish.The inventive method makes the biochip obtained, and has
The most anti-non-specific.
Accompanying drawing explanation
Fig. 1 is that chip surface sample prints array of figure；
Fig. 2 is each sample point and IgG protein binding signal figure.
Detailed description of the invention
The synthetic method of a kind of macromolecule mother solution, comprises the steps:
1) OEGMA and methacrylic acid derivative monomer are dissolved in polar solvent, remove bubble removing, keep away under protection gas shielded
Light stirs, and obtains monomer mixed solution A, standby；
2) appropriate complex ligand and catalyst transition metal halide are dissolved in the water, obtain solution B；
3) by monomer mixed solution A and solution B mix homogeneously under protection gas shielded, to reactant liquor variable color；
4) in reactant liquor, add weak reductant, be stirring evenly and then adding into initiator initiated polymerization；
5) after reaction completely, by oxygen-containing gas, aoxidize solution, obtain macromolecule mother solution.
Polar solvent effect is effectively to dissolve OEGMA and methacrylic acid derivative monomer, and its dissolubility is high favourable
Carrying out in liquid phase synthesis.Conventional polar solvent can be the mixed liquor of first alcohol and water, as methanol mixes with water equal-volume
The mixed liquor arrived.The solution that polarity is big can improve Polymer Synthesizing speed, reduces final high molecular molecular size range, methanol
Add, be on the one hand to reduce reaction rate, improve molecular weight, be on the other hand to increase system to high polymer monomer dissolubility.
As the further improvement of above-mentioned synthetic method, complex ligand is bipyridyl.Particularly, bipyridyl is 2, and 2-joins
Pyridine.
As the further improvement of above-mentioned synthetic method, transition metal halide is copper halide.Particularly, copper halide is
CuCl2、CuBr2Halogenide Deng stable existence.
Weak reductant is weak reductant commonly used in the art, refer under this experimental system can optionally by cupric from
Son is reduced into the reducing agent of univalent copper ion, makes to exist in reaction system univalent copper ion and bivalent cupric ion simultaneously, is beneficial to height
The synthesis of molecule.And bivalent cupric ion can be reduced into copper simple substance by the strong reductants such as the strongest reducing agent such as Pd-C, NaH,
Be not suitable for this reaction system.As the further improvement of above-mentioned synthetic method, weak reductant is selected from ascorbic acid, glucose or pungent
Acid stannous.In view of the convenience etc. used, weak reductant most preferably ascorbic acid.
As the further improvement of above-mentioned synthetic method, initiator is preferably sulfur-bearing initiator, selected from single sulfydryl halo sulfur
Alcohol, single mercapto-polyglycol oligomer, double sulfydryl halogenated thiols, double mercapto-polyglycol oligomer etc..That concrete can be HS-
(CH2)6-OC (O)-IzoButyrate-Br, HS-(CH2)12OC (O)-IzoButyrate-Br, HS-(CH2)18OC(O)-
IzoButyrate-Br etc..Sulfur-bearing initiator, during participating in macromolecular polymerization reaction, can introduce S in macromolecule simultaneously
Key, it is simple to macromolecule is fixed on chip surface in follow-up reaction.
As the further improvement of above-mentioned synthetic method, the mol ratio of OEGMA and methacrylic acid derivative monomer be (1:
4)～(4:1).By the ratio of both regulations, can be with the space between the final molecular size range of accuracy controlling macromolecule and macromolecule
Size.
Particularly, the molecular weight of OEGMA monomer is between 300Da～2000Da.
As the further improvement of above-mentioned synthetic method, the high molecular number-average molecular weight of synthesis is 500,000～5,000,000.Real
Test the macromolecule in result shows this molecular weight ranges, the anti-non-specific of biochip can be effectively improved.
As the further improvement of above-mentioned synthetic method, methacrylic acid derivative monomer is the methacrylic acid of hydroxyl
Derivatives monomer.The methacrylic acid derivative monomer of hydroxyl can be hydroxyethyl methylacrylate, hydroxyethyl methyl propylene
Acid hydroxyl ethyl ester, carboxymethylmethyl 2-(Acryloyloxy)ethanol, carboxyethyl methylphosphinate 2-(Acryloyloxy)ethanol etc..
The manufacture method of a kind of 3D biochip, including using on modifying agent macromolecule in above-mentioned macromolecule mother solution
Introduce anchoring group, obtain derivatization macromolecule mother solution；Take derivatization macromolecule mother solution afterwards to be laid on carrier, after reaction completely
Clean and remove loose macromolecule, be dried to obtain 3D biochip.
As the further improvement of above-mentioned manufacture method, anchoring group is selected from-COOH ,-NH2,-OH, epoxy radicals ,-OCH3,-
COCNH2, cyano group, alkynyl, azido.The introducing of anchoring group can use existing method to carry out, such as Jia P, Tang Y C,
Ning X, et al. Covalently derivatized NTA microarrays on porous silicon for
multi-mode detection of His-tagged proteins [J]. Science China, 2011, 54(3):
526-535. realize Deng disclosed method.
As the further improvement of above-mentioned manufacture method, carrier is for having nanometer gold, nanometer silver, nanometer chromium, three oxidations two
Aluminum or the chip of titanium dioxide surface.
The present invention is abbreviation commonly used in the art, if any difference, is as the criterion with noun abbreviation defined under:
OEGMA: oligomeric ethylene glycol methyl ether methacrylate
HEMA: hydroxyethyl methylacrylate
BPY:2,2-bipyridyl
DMAP:4-dimethylamino naphthyridine
DMF: dimethylformamide
EDC:1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimides
NHS:N-N-Hydroxysuccinimide.
Below in conjunction with embodiment, further illustrate technical scheme.
The synthesis of macromolecule mother solution 1:
1) 10mmol OEGMA 500 and 10mmol HEMA is dissolved in MeOH(25 mL) and the mixed solution of water (25 mL)
In, after sonic oscillation removes bubble removing, lucifuge, it is passed through nitrogen lower magnetic agitation 30 min of protection, obtains monomer mixed solution A；
2) by 32 mg BPY, 2.5 mL 0.04 M CuCl2Aqueous solution uniform, obtain solution B；
3) under continuing nitrogen protection, injecting in solution A by syringe holder B solution, magnetic agitation 5 min, in light blue；
4) draw 1.5 mL 0.04 M aqueous ascorbic acids with syringe to be expelled in above-mentioned reaction solution, magnetic agitation 5
Min solution is brown；
5) 1mL 1 mM sulfur-bearing initiator HS-(CH is drawn with syringe2)6-OC (O)-IzoButyrate-Br is expelled to above-mentioned
In solution, solution is brown, after reacting 20 h, is passed through air oxidation solution, obtains macromolecule mother solution.
The synthesis of macromolecule mother solution 2:
1) 10mmol OEGMA 500 is dissolved in MeOH(25 mL) and the mixed solution of water (25 mL) in, sonic oscillation remove
After bubble, lucifuge, it is passed through nitrogen lower magnetic agitation 30 min of protection, obtains monomer mixed solution A；
2) by 32 mg BPY, 2.5 mL 0.04 M CuCl2Aqueous solution uniform, obtain solution B；
3) under continuing nitrogen protection, injecting in solution A by syringe holder B solution, magnetic agitation 5 min, in light blue；
4) draw 1.5 mL 0.04 M aqueous ascorbic acids with syringe to be expelled in above-mentioned reaction solution, magnetic agitation 5
Min solution is brown；
5) 1mL 1 mM sulfur-bearing initiator HS-(CH is drawn with syringe2)6-OC (O)-IzoButyrate-Br is expelled to above-mentioned
In solution, solution is brown, after reacting 20 h, is passed through air oxidation solution, obtains macromolecule mother solution 2.
The synthesis of macromolecule mother solution 3:
1) 2.5mmol OEGMA 500 and 10mmol HEMA is dissolved in MeOH(25 mL) and the mixed solution of water (25 mL)
In, after sonic oscillation removes bubble removing, lucifuge, it is passed through nitrogen lower magnetic agitation 30 min of protection, obtains monomer mixed solution A；
2) by 32 mg BPY, 2.5 mL 0.04 M CuCl2Aqueous solution uniform, obtain solution B；
3) under continuing nitrogen protection, injecting in solution A by syringe holder B solution, magnetic agitation 5 min, in light blue；
4) draw 1.5 mL 0.04 M aqueous ascorbic acids with syringe to be expelled in above-mentioned reaction solution, magnetic agitation 5
Min solution is brown；
5) 1mL 1 mM sulfur-bearing initiator HS-(CH is drawn with syringe2)6-OC (O)-IzoButyrate-Br is expelled to above-mentioned
In solution, solution is brown, after reacting 20 h, is passed through air oxidation solution, obtains macromolecule mother solution 3.
The synthesis of macromolecule mother solution 4:
1) 10mmol OEGMA 500 and 2.5mmol HEMA is dissolved in MeOH(25 mL) and the mixed solution of water (25 mL)
In, after sonic oscillation removes bubble removing, lucifuge, it is passed through nitrogen lower magnetic agitation 30 min of protection, obtains monomer mixed solution A；
2) by 32 mg BPY, 2.5 mL 0.04 M CuCl2Aqueous solution uniform, obtain solution B；
3) under continuing nitrogen protection, injecting in solution A by syringe holder B solution, magnetic agitation 5 min, in light blue；
4) draw 1.5 mL 0.04 M aqueous ascorbic acids with syringe to be expelled in above-mentioned reaction solution, magnetic agitation 5
Min solution is brown；
5) 1mL 1 mM sulfur-bearing initiator HS-(CH is drawn with syringe2)6-OC (O)-IzoButyrate-Br is expelled to
Stating in solution, solution is brown, after reacting 20 h, is passed through air oxidation solution, obtains macromolecule mother solution 4.
The synthesis of macromolecule mother solution 5:
1) 10mmol HEMA is dissolved in MeOH(25 mL) and the mixed solution of water (25 mL) in, sonic oscillation removes bubble removing
After, lucifuge, it is passed through nitrogen lower magnetic agitation 30 min of protection, obtains monomer mixed solution A；
2) by 32 mg BPY, 2.5 mL 0.04 M CuCl2Aqueous solution uniform, obtain solution B；
3) under continuing nitrogen protection, injecting in solution A by syringe holder B solution, magnetic agitation 5 min, in light blue；
4) draw 1.5 mL 0.04 M aqueous ascorbic acids with syringe to be expelled in above-mentioned reaction solution, magnetic agitation 5
Min solution is brown；
5) 1mL 1 mM sulfur-bearing initiator injection HS-(CH is drawn with syringe2)6-OC (O)-IzoButyrate-Br is to above-mentioned
In solution, solution is brown, after reacting 20 h, is passed through air oxidation solution, obtains macromolecule mother solution 5.
Synthesize high molecular molecular weight mainly to be affected by following condition: 1, high polymer monomer ratio；2, the response time；3, instead
Answer temperature；4, ratio of initiator.In above-mentioned experiment, constant by controlling other reaction conditions, only change high polymer monomer ratio
Example, can accurately control to obtain the macromolecule mother solution (see Table 1) of different molecular weight.
Table 1, the impact on molecular weight of the raw material mixing ratio
Under the same terms, the prolongation response time can obtain the macromolecule of higher molecular weight；Reaction temperature the highest reaction molecular amount
Being distributed the most extensive, molecular weight is the least；Ratio of initiator is the biggest, and synthesis molecular weight polymeric is the least, otherwise, then molecular weight is the biggest.
Those skilled in the art can also select accurately to control synthesis by control response time, reaction temperature, ratio of initiator
High molecular molecular weight.
High molecular modification:
1) room temperature preparation 1g fourth two acetic anhydride, 0.75g DMAP, 100ml DMF souring soln, stirring reaction 1 hour, reacted
Acidifying solution；
2) take above-mentioned macromolecule mother solution 100ml and reaction acidifying solution 100ml, 24 hs are stirred at room temperature, remove solvent DMF, obtain dry
Sample；
3) drying sample being dissolved in 100ml ethanol, preparation obtains derivatization macromolecule mother solution.
Or use this area method known to other, such as Jia P, Tang Y C, Ning X, et al.
Covalently derivatized NTA microarrays on porous silicon for multi-mode
Detection of His-tagged proteins [J]. Science China, the public affairs such as 2011,54 (3): 526-535.
That opens is modified macromolecule, the anchoring group required for introducing.
Naked gold surface self assembly macromolecule membrane:
Alcohol flushing chip three times, cleans 5 minutes with plasma cleaner after drying, dries up naked gold glass-chip, standby；
Take the derivatization macromolecule mother solution of the above-mentioned synthesis of 1ml, be layered on naked gold glass-chip, react 5 minutes；
Through ethanol, after water respectively cleans once, nitrogen dries up, and the synthesis of the most naked gold surface self assembly macromolecule membrane is complete, obtains
3D naked gold biochip.
Naked silver surface self assembly macromolecule membrane:
1) alcohol flushing chip three times, clean 5 minutes with plasma cleaner after drying, dry up silver glass-chip, standby；
2) take the derivatization macromolecule mother solution of the above-mentioned synthesis of 1ml, be layered on naked silver glass-chip, react 5 minutes；
3) through ethanol, after water respectively cleans once, nitrogen dries up, and the synthesis of so far silver surface self assembly macromolecule membrane is complete,
To 3D silver biochip.
Aluminium sesquioxide surface self-organization macromolecule membrane:
1) alcohol flushing chip three times, clean 5 minutes with plasma cleaner after drying, dry up aluminium sesquioxide glass-chip,
Standby；
2) take the derivatization macromolecule mother solution of the above-mentioned synthesis of 1ml, be layered on aluminium sesquioxide glass-chip, react 5 minutes；
3) through ethanol, after water respectively cleans once, nitrogen dries up, and so far aluminium sesquioxide surface self-organization macromolecule membrane has synthesized
Finish, obtain 3D aluminium sesquioxide biochip.
3D naked gold biochip is for polypeptide, antibody, proteopexy
1) above-mentioned naked gold self assembly macromolecule chip is activated by the activating solution of the 10mM EDC/NHS using mol ratio to be 1:1
Process 30 points
2) pure water Rapid Cleaning, nitrogen dries up, then with Q Array Mini high flux point model machine, to polypeptide, antibody, albumen sample
Product carry out point sample process according to pre-set program, keep deposition process to be in moisture state；
3) use the 1M aqueous ethanolamine of pH=8.5 that chip carries out sealing treatment, react half an hour；
4) using 10XPBS, 2XPBS, 1XPBS the most successively, cleaning each to chip 15 minutes, after nitrogen dries up, 4 DEG C of refrigerators are protected
Depositing standby, so far, polypeptide, antibody, the fixing of protein sample completes.
Macromolecule modified naked gold chip detects for antibody-antigene
Experimental group:
According to array shown in Fig. 1,1mg/mL Anti-IGG antibody protein is beaten by Q Array Mini high flux point model machine
Print naked gold chip surface after activation.Hatch through liquid, close, after cleaning, after adding the micro-fluidic COVER of Plexera company,
On Plexera HT instrument, test according to Standard test programme.
Matched group:
According to array shown in Fig. 1,1mg/mLBSA antibody protein is printed upon by Q Array Mini high flux point model machine live
Naked gold chip surface after change.Hatch through liquid, close, after cleaning, after adding the micro-fluidic COVER of Plexera company,
On Plexera HT instrument, test according to Standard test programme.Selected buffering liquid is 1 × PBST (0.05%T-20),
Liquid of living again is 1:200 H3PO4Aqueous solution, the flow through sample degree of depth is 10 μ g/ml IGG antibody proteins.
Blank group:
According to array shown in Fig. 1,1mg/mL1XPBS solution is printed upon by Q Array Mini high flux point model machine activation
After naked gold chip surface.Hatch through liquid, close, after cleaning, after adding the micro-fluidic COVER of Plexera company,
On Plexera HT instrument, test according to Standard test programme.Selected buffering liquid is 1 × PBST (0.05%T-20),
Liquid of living again is 1:200 H3PO4Aqueous solution, the flow through sample degree of depth is 10 μ g/ml IgG antibody albumen.
Detection: selected buffering liquid is 1XPBST (0.05%T-20), and liquid of living again is 1:200 H3PO4Aqueous solution, stream
Logical sample depth is 10 μ g/ml IGG antibody proteins.Successively on chip, circulate liquid (3 μ l/s, 300s) of living again, buffer (2
μ l/s, 300s), 10 μ g/ml IGG samples (2 μ l/s, 300s), obtain each sample by PlexeraDE.exe software analysis
Point and circulation antibody protein binding signal, result is as shown in Figure 2.
Experimental group: Anti-IGG sample spot has obvious binding signal, signal magnitude is 3 Au units, and affinity size is
10-15M；
Matched group: BSA sample spot is without obvious binding signal, and signal magnitude is 0 Au unit, and affinity size is 0.
Blank group: sample spot is without obvious binding signal, and signal magnitude is 0 Au unit, and affinity size is 0.
Matched group and blank group are all without obvious binding signal.Contrast three groups of data, it is known that the macromolecule that this method makes is repaiied
The naked gold chip (3D naked gold biochip) of decorations has stronger anti-non-specific performance, can be used for what antibody-antigene interacted
High flux screening.
Claims (10)
1. a synthetic method for macromolecule mother solution, comprises the steps:
1) OEGMA and methacrylic acid derivative monomer are dissolved in polar solvent, go bubble removing, protection gas to protect 2) protect down
Lucifuge stirs, and obtains monomer mixed solution A, standby；
3) appropriate complex ligand and catalyst transition metal halide are dissolved in the water, obtain solution B；
4) by monomer mixed solution A and solution B mix homogeneously under protection gas shielded, to reactant liquor variable color；
5) in reactant liquor, add weak reductant, be stirring evenly and then adding into initiator initiated polymerization；
6) after reaction completely, by oxygen-containing gas, aoxidize solution, obtain macromolecule mother solution.
Synthetic method the most according to claim 1, it is characterised in that: complex ligand is bipyridyl.
Synthetic method the most according to claim 1, it is characterised in that: transition metal halide is copper halide.
Synthetic method the most according to claim 1, it is characterised in that: weak reductant is selected from ascorbic acid, glucose or pungent
Acid stannous.
Synthetic method the most according to claim 1, it is characterised in that: initiator is selected from single sulfydryl halogenated thiols, single sulfydryl
Polyethylene Glycol oligomer, double sulfydryl halogenated thiols, double mercapto-polyglycol oligomer.
Synthetic method the most according to claim 1, it is characterised in that: rubbing of OEGMA and methacrylic acid derivative monomer
Your ratio is (1:4)～(4:1).
Synthetic method the most according to claim 1, it is characterised in that: the high molecular number-average molecular weight of synthesis be 500,000～
5000000.
Synthetic method the most according to claim 1, it is characterised in that: methacrylic acid derivative monomer is the first of hydroxyl
Base acrylic acid derivative monomer.
9. a manufacture method for 3D biochip, including using modifying agent at the high score described in claim 1～8 any one
Introduce anchoring group on macromolecule in primary and secondary liquid, obtain derivatization macromolecule mother solution；Take derivatization macromolecule mother solution paving afterwards
On carrier, clean after reaction completely and remove loose macromolecule, be dried to obtain 3D biochip.
Manufacture method the most according to claim 9, it is characterised in that: anchoring group is selected from-COOH ,-NH2,-OH, epoxy
Base ,-OCH3,-COCNH2, cyano group, alkynyl, azido.
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