CN109917139A - A kind of GMR chip and the magnetosensitive immune detection card including it - Google Patents
A kind of GMR chip and the magnetosensitive immune detection card including it Download PDFInfo
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- CN109917139A CN109917139A CN201910255571.2A CN201910255571A CN109917139A CN 109917139 A CN109917139 A CN 109917139A CN 201910255571 A CN201910255571 A CN 201910255571A CN 109917139 A CN109917139 A CN 109917139A
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- maleic anhydride
- gmr
- chip
- detection
- magnetosensitive
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- 238000001514 detection method Methods 0.000 title claims abstract description 46
- FPYJFEHAWHCUMM-UHFFFAOYSA-N maleic anhydride Chemical compound O=C1OC(=O)C=C1 FPYJFEHAWHCUMM-UHFFFAOYSA-N 0.000 claims abstract description 44
- 229920006254 polymer film Polymers 0.000 claims abstract description 14
- 238000002331 protein detection Methods 0.000 claims abstract description 7
- 238000000034 method Methods 0.000 claims description 24
- 238000007789 sealing Methods 0.000 claims description 24
- 238000006243 chemical reaction Methods 0.000 claims description 23
- 239000004205 dimethyl polysiloxane Substances 0.000 claims description 23
- 235000013870 dimethyl polysiloxane Nutrition 0.000 claims description 23
- CXQXSVUQTKDNFP-UHFFFAOYSA-N octamethyltrisiloxane Chemical compound C[Si](C)(C)O[Si](C)(C)O[Si](C)(C)C CXQXSVUQTKDNFP-UHFFFAOYSA-N 0.000 claims description 23
- 238000004987 plasma desorption mass spectroscopy Methods 0.000 claims description 23
- 229920000435 poly(dimethylsiloxane) Polymers 0.000 claims description 23
- 238000004528 spin coating Methods 0.000 claims description 20
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- 238000003860 storage Methods 0.000 claims description 13
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- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- 239000004698 Polyethylene Substances 0.000 description 2
- 229910000831 Steel Inorganic materials 0.000 description 2
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 2
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- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 2
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Landscapes
- Investigating Or Analyzing Materials By The Use Of Magnetic Means (AREA)
Abstract
The present invention discloses a kind of GMR chip and the magnetosensitive immune detection card including it, is related to protein detection technology field.The surface of the GMR chip includes the maleic anhydride polymer film layer for capturing antibody, which can be effectively secured to chip surface for antibody by amidation.The present invention also provides a kind of magnetosensitive immune detection card for Protein Detection including above-mentioned GMR chip, the detection sensitivity of the detection card are high, easy to operate.
Description
Technical field
The present invention relates to protein detection technology fields.More particularly, to a kind of GMR chip and including its magnetosensitive it is immune
Detection card.
Background technique
Magnetosensitive immuno analytical method uses indirect method, is based on giant magnetoresistance (GMR, Giant Magneto Resistance) core
The giant magnetoresistance effect of piece detects nanometer magnetic conduction iron shot (immunomagnetic beads) content, contains so that quantitative resolution goes out antibody to be checked in sample
Amount.GMR chip base main component is monocrystalline silicon, and monocrystalline silicon surface is very weak to protein adsorption ability, it is therefore desirable to chip list
Face is chemically modified, and improves the ability of chip surface adhesion protein.When carrying out surface chemical modification, needs to solve three and ask
Topic, first, GMR chip surface adhesion protein amount need to be improved；Second, GMR chip electromagnetic device need to be avoided for a long time by organic molten
Liquid corrodes；Third need to reduce probe magnetic particle and improve detection sensitivity at a distance from electromagnetic device magnetosensitive layer.
Antibody is fixed on carrier to be broadly divided into covalent method and non-covalence method, and non-covalence method is inhaled by hydrophobic effect, electrostatic
The effects of attached, hydrogen bond, antibody was fixed on carrier surface by power, and the efficiency of this method sessile antibody is not often high, but due to this
Method operation is fairly simple, is still widely used at present.Antibody is connected to carrier surface by chemical bond by covalent method, relative to non-
Covalent method, this method are higher to the fixed efficiency of antibody, but reaction process is harsher.
Biotin-avidin is common amplification system in immunoassay, and antibody is normally used for the official of biotin modification
Can roll into a ball mainly has amino, carbonyl, sulfydryl.In the above-mentioned methods, method of modifying of the biotin on antibody amino groups be operate it is most simple
It is single, and on antibody biotin labelled amount also highest, signal amplification is most strong, but since Fab fragments are there are a large amount of amino,
Amido modified method may change monoclonal antibody conformation, influence the combination of antibody and antigen.Carbonyl, sulfydryl reactivity biotin
General modification antibody Fc fragment, both methods may be implemented biotin directional at-tachment and quantitative fixation, and biotin is fixed
Position and antigen binding regions farther out, will not to antigen-antibody combine interfere.But during carbonyl modified, need to use
To Strong oxdiative reagent, have a certain impact to the activity of antibody, and sulfydryl modification needs the disconnection antibody hinge region of selectivity
Disulfide bond, it is therefore necessary to use suitable reducing agent.
Microfluidic chip technology refers to accurately controlling using microchannel and handling minute yardstick fluid, thus on microchip
Realize that the multiple functions such as sample introduction, dilution, mixing, reaction, detection, most prominent advantage are only to need a small amount of sample or biological sample
Product can efficiently quickly finish the detection of various microanalysis, and have pollution less, low cost, sample size demand be small, detection reagent
The few and high-throughput advantage of consumption.Microfluidic chip technology is quickly grown in recent years, is had a wide range of applications in analysis field.Mirror
In micro-fluidic chip have the advantages such as large sample size parallel work-flow are completed at the same time under miniature scale, by microfluidic chip technology with
Immunoassay combines, and is the technology that new development in recent years is got up, and substantially improves traditional immunization analysis performance.Such as patent
Microchannel is applied to different immunoassays and put down by CN201710957817, CN201710542327,201810509672
Platform greatly extends the application of immunoassay.
Therefore, GMR chip, magnetic particle marker and microchannel and double antibodies sandwich immunoassay method, production one are merged
Money polyprotein joint-detection platform has great importance.
Summary of the invention
It is an object of the present invention to provide a kind of GMR chip, which can be effectively solid by antibody by amidation
It is scheduled on chip surface.
It is immune that it is another object of the present invention to provide a kind of magnetosensitives for Protein Detection including above-mentioned GMR chip
Detection card, the detection sensitivity of the detection card are high, easy to operate.
In order to achieve the above objectives, the present invention adopts the following technical solutions:
A kind of GMR chip, the surface of the GMR chip include the maleic anhydride polymer film for capturing antibody
Layer.
Preferably, the maleic anhydride polymer film layer with a thickness of 6-42nm.
Preferably, the maleic anhydride polymer film layer is existed by the method modification of spin coating maleic anhydride polymer solution
Chip surface.
Preferably, the process of spin coating includes: 500-1500 revs/min of spin coating 6s；Then 5000-10000 revs/min of spin coating 20s.
Preferably, it is total to be selected from polyethylene maleic anhydride copolymer, polystyrene maleic anhydride for the maleic anhydride polymer
Polymers, polystyrene graft maleic anhydride, Research of Grafting Malaic Anhydride Onto Polyethylene, polypropylene grafted maleic anhydride and polyisoprene connect
One or more of branch maleic anhydride.
Preferably, the maleic anhydride polymer is polystyrene graft maleic anhydride.
Preferably, the GMR chip includes several reaction zones, and each reaction zone can a kind of albumen of independent detection.
The present invention also provides application of described in any item GMR chips in polyprotein joint-detection as above.
According to the second object of the invention, the present invention provides a kind of magnetosensitive immune detection card for Protein Detection, packet
Include described in any item GMR chips as above.
It preferably, from top to bottom successively include top cover, PDMS sealing cover, reagent storage layer, PDMS sealing film, printed circuit
Plate and bottom cover；The GMR chip is pasted on the printed circuit board and is electrically connected with printed circuit board.
Beneficial effects of the present invention are as follows:
The Malaysia of one layer of only 6-42nm thickness has been modified on a kind of GMR chip provided by the invention, surface by spin coating technique
Anhydride polymer film layer significantly improves the ability of chip capture antibody, greatly reduces magnetic particle at a distance from magnetosensitive layer,
Spin coating process only needs 26s that can complete, and organic solvent is avoided to corrode chip for a long time.
The present invention also provides a kind of magnetosensitive immune detection card including above-mentioned GMR chip, which has detection sensitive
The advantage that degree is high, detection range is wide, detection time is short, easy to operate, sample volume used is small, single detection project is at low cost,
Suitable for situation of all-level hospitals and R&D institution, and magnetosensitive immune detection card has the ability to expand to the joint inspection of greater protein marker
In survey.
Detailed description of the invention
Specific embodiments of the present invention will be described in further detail with reference to the accompanying drawing.
Fig. 1 shows magnetosensitive immune detection card GMR chip structure figure in the embodiment of the present invention 1.
101.CEA coated antibody fixes position；102.NSE coated antibody fixes position；103.ProGRP coated antibody is solid
Positioning is set；104.CYFRA21-1 coated antibody fixes position；105.SCC coated antibody fixes position.
Fig. 2 (a) shows influence of the polystyrene graft maleic anhydride concentration to film thickness, and Fig. 2 (b) shows polystyrene graft
Influence of the maleic anhydride concentration to magnetic response signal.
Fig. 3 (a) shows when the embodiment of the present invention 3 detects nonantigenic magnetic particle in the capture situation of chip surface；Fig. 3 (b)
The detection of the embodiment of the present invention 3 capture situation of magnetic particle in chip surface when having antigen is shown.
Fig. 4 shows the magnetosensitive immune detection card microchannel structure figure of 3 lung cancer, 5 tumor markers of the embodiment of the present invention.
401. chips fix position；402. magnetic particle storage locations；403. biotinylation labelled antibody storage locations；404.
Magnetic particle redissolves liquid；405. sample solution；406. magnetic particle level monitorings；407. sample level monitorings；408. cleaning solution.
Fig. 5 shows the standard curve of five projects of CEA, NSE, CYFRA21-1, ProGRP, SCC of embodiment 5.
Specific embodiment
In order to illustrate more clearly of the present invention, the present invention is done further below with reference to preferred embodiments and drawings
It is bright.Similar component is indicated in attached drawing with identical appended drawing reference.It will be appreciated by those skilled in the art that institute is specific below
The content of description is illustrative and be not restrictive, and should not be limited the scope of the invention with this.
In the prior art, GMR chip base main component is monocrystalline silicon, and monocrystalline silicon surface is very weak to protein adsorption ability,
Therefore it needs to be chemically modified chip surface, to improve the ability of chip surface adhesion protein.
Present invention firstly provides a kind of GMR chips, which is characterized in that the surface of the GMR chip includes for capturing
The maleic anhydride polymer film layer of antibody.The maleic anhydride polymer film layer can be sent out by the amino of anhydride group and antibody
Raw amidation process, is effectively fixed in chip surface for coated antibody.
Further, the maleic anhydride polymer film layer with a thickness of 6-42nm, such as can for 6nm, 12nm,
19nm,32nm,42nm；Preferably, maleic anhydride polymer film layer with a thickness of 19nm.Maleic acid in the thickness range
Anhydride polymer film layer can not only play the function of capture antibody, but can reduce probe magnetic particle and electromagnetic device magnetosensitive layer away from
From improving the sensitivity of detection.
Preferably, the maleic anhydride polymer film layer is existed by the method modification of spin coating maleic anhydride polymer solution
Chip surface.It is poly- that those skilled in the art can also modify maleic anhydride in chip surface by other common methods of this field
Object film layer is closed, the present invention is not particularly limited this.
Further, the process of spin coating includes: 500-1500 revs/min of spin coating 6s, for example, herein revolving speed can for 500,
800,1000,1200,1500；Then 5000-10000 revs/min of spin coating 20s again, for example, herein revolving speed can for 5000,6000,
8000,10000.Carrying out spin coating using the spin coating method only needs 26s that can complete, and organic solvent is avoided to corrode chip for a long time.
Those skilled in the art can also adjust spin coating revolving speed and time conditions according to the actual situation, be able to satisfy and coat symbol in chip surface
Close the present invention claims polymer film layer.
The volume of the maleic anhydride polymer solution used when spin coating is 2mL, and the concentration of maleic anhydride polymer is
0.2%-3%, such as can be 0.2%, 0.5%, 1%, 2% and 3%, it is preferable that the concentration of maleic anhydride polymer is
1%.
The maleic anhydride polymer is selected from polyethylene maleic anhydride copolymer, polystyrene maleic anhydride copolymer, gathers
Styrene-grafted maleic anhydride, Research of Grafting Malaic Anhydride Onto Polyethylene, polypropylene grafted maleic anhydride and polyisoprene are grafted Malaysia
One or more of acid anhydrides.Further, the maleic anhydride polymer is polystyrene graft maleic anhydride.
Carrying out solvent used in polystyrene graft maleic anhydride solution used when spin coating is benzene,toluene,xylene, four
Hydrogen furans, acetone, ethyl acetate, chloroform, methylene chloride, hexamethylene, cyclohexanone, trichloro ethylene, naphthane, one in decahydronaphthalene
Kind is several, it is preferable that the solvent is dimethylbenzene.
Preferably, the GMR chip includes several reaction zones, and each reaction zone can a kind of albumen of independent detection.For example, real
GMR chip described in the application of border can be equipped with 40 reaction zones, the column of 4 rows × 10, each reaction zone can one albumen of independent detection,
It at most can detect 40 albumen；It, can detection 5 altogether if detecting a kind of lung cancer marker using 2 column.
The present invention also protects application of described in any item GMR chips in polyprotein joint-detection as above.This field skill
Art personnel, can also be by multiple reaction zones for being arranged thereon it is found that above-mentioned GMR chip both can be used for the detection of single albumen
Multiple protein is detected simultaneously, can play the role of high-throughput and saves sample liquid.
The present invention also provides a kind of magnetosensitive immune detection cards for Protein Detection, including as above described in any item
GMR chip.Further, the magnetosensitive immune detection card successively includes top cover, PDMS sealing cover, reagent storage from top to bottom
Layer, PDMS sealing film, printed circuit board and bottom cover；The GMR chip is pasted on the printed circuit board and and printed circuit
Plate electrical connection.Top cover and PDMS sealing cover, reagent storage layer form liquid storage tank.PDMS sealing cover is acted on because of tool elasticity in pressure
Under liquid can be driven to flow in microchannel.There are three liquid storage tanks on reagent storage layer top: sample cell, cleaning liquid pool and magnetic
Three liquid storage tanks are connected on GMR chip by grain dissolution liquid pool, design of micro fluidic channels in the bottom of reagent storage layer, microchannel
The reaction chamber of side.PDMS sealing film is for sealing microfluidic channel.GMR chip is pasted on printed circuit board, it will using aluminum steel
Chip and printed circuit board interconnect, with the switch of level detection on printed circuit board, it is ensured that microfluid is accurately had
The control of effect.Bottom cover is the pedestal of entire test card, contains positioning column, bottom cover, reagent storage layer and top cover are by ultrasonic welding
One, realizes the complete seal of entire test card microfluidic channel.
GMR chip, magnetic particle marker and microchannel and double antibodies sandwich immunoassay method, common inspection are used when comprehensive
When surveying albumen, GMR chip, the labelled antibody of biotin modification and Streptavidin magnetic particle are individually positioned in the corresponding of microchannel
Position.When being tested, sample solution, magnetic particle solution, cleaning solution are transported to chip using push type active microfluidic system
Surface, the specific steps are as follows:
(1) the PDMS sealing cover above sample cell is opened, 50 μ L sample solutions are added in sample cell.
(2) the PDMS sealing cover above sample cell is pressed, sample solution is injected into sample level sensing position, then,
The PDMS sealing cover above pressing sample cell shakes sample solution repeatedly in labelled antibody freeze-drying area up and down, it is ensured that labelled antibody
It is completely dissolved in sample solution.
(3) continue to press PDMS sealing cover, labelled antibody is made to be injected into GMR chip list together with the mixed solution of sample
The labelled antibody of biotin modification is consolidated by antigen in coated antibody-sample-labelled antibody sandwich immunoassay reaction pattern in face
It is scheduled on chip surface.
(4) PDMS sealing cover above pressing cleaning solution, injects cleaning solution, extra antigen is washed away with labelled antibody.
(5) pressing magnetic particle answers superjacent PDMS sealing cover, and magnetic particle redissolution liquid is injected into magnetic particle level sensing
Position, shaking magnetic particle redissolution liquid repeatedly is completely dissolved the magnetic particle of freeze-drying.
(6) continue pressing magnetic particle and answer superjacent PDMS sealing cover, magnetic particle solution is injected into chip surface, is passed through
The magnetic particle of biotin-avidin system, Streptavidin modification is captured to chip surface, under the influence of external magnetic field,
Magnetic particle generates derivative magnetic field, causes GMR reaction zone resistance variations.
Labelled antibody uses 1- biotin acylamino -4- [4`- (methylycaconitine) cyclohexane carboxamide] butane, N- iodine
Acetyl-N- biotin hexamethylene diamine, two polyethylene glycol biotin ester of maleimide, six polyethylene glycol biotin of maleimide
One or more of ester, 11 polyethylene glycol biotin ester of maleimide modification, it is preferable that the biotin is Malaysia acyl
11 polyethylene glycol biotin ester of imines.
The labelled antibody uses in dithiothreitol (DTT), 2 mercapto ethanol, Mercaptamine, three (2- carboxyethyl) phosphines
One or more of reducing agents selectivity disconnection its hinge area disulfide bond, generate have free sulfhydryl groups incomplete antibody, preferably
Ground, the reducing agent are Mercaptamine.Labelled antibody uses sulfydryl reactivity biotin modification, realizes biotin confrontation
Body orientation and quantitative modification.The sulfydryl reactivity biotin introduces a large amount of hydrophilic radical, increases labelled antibody
Hydrophily prevents labelled antibody from reuniting in the solution.
Sample solution, magnetic particle solution, the cleaning solution of test card use push type active microfluidic system as described above
It is transported to chip surface, participates in double antibodies sandwich immune response and magnetic nano particle reaction.
1 coated antibody of embodiment is fixed on GMR chip surface
(1) polystyrene graft maleic anhydride is dissolved in dimethylbenzene, it is that 1.0% poly- polystyrene connects that preparing, which becomes concentration,
Branch maleic anhydride solution, wafer is placed on sol evenning machine, starts sol evenning machine, and it is molten that 2mL polystyrene graft maleic anhydride is added dropwise
Liquid, it is ensured that for solution at 800 revs/min at a slow speed, the time is to drip off in 6s, then continues to rotate 20s at 6000 revs/min, obtains thickness
The polystyrene graft Maleic Anhydride Films layer of about 19nm.
(2) wafer after spin coating is cut into required GMR chip using laser cutting machine, and GMR chip is pasted
GMR chip is realized with printed circuit board interconnection using aluminum steel and is electrically connected by the corresponding position of printed circuit board.
(3) the present embodiment detects lung cancer marker in 5 using GMR chip provided by the invention and magnetosensitive immune detection card,
The lung cancer marker includes: carcinomebryonic antigen (CEA), neuronspecific enolase (NSE), cytokeratin fragment 19
(CYFRA21-1) and gastrin-releasing peptide precursor (ProGRP) and squamous cell carcinoma-related antigen (SCC).50 μ are prepared respectively
The CEA of g/mL, NSE, ProGRP, SCC, CYFRA21-1 coated antibody solution.It sorts according to scheduled array, every two column are fixed
Five kinds of coated antibodies are fixed to chip surface using point sample instrument by a kind of coated antibody.
(4) the fixation position of GMR chip and five kinds of coated antibodies described in is as shown in Figure 1.
2 labelled antibody biotin modification of embodiment
(1) in PH=7.2, it is added 1mg labelled antibody in the PBS-EDTA solution of the Mercaptamine containing 50mM, 37
DEG C reaction 90min.
(2) reaction solution in (1) is cooled to room temperature, using the ultrafiltration of PBS-EDTA solution 5 times of PH=7.0, is removed
Unreacted Mercaptamine.
(3) the labelled antibody concentration of the reaction solution in (2) is adjusted to 2mg/ml using the PBS solution of PH=7.0, is added
Enter the 11 polyethylene glycol biotin ester solution of maleimide that 13.3 μ L concentration are 20mM, 4 DEG C of reaction 2h.
(4) use the PBS solution of PH=7.4 that the reaction solution ultrafiltration in (3) three times, is removed unreacted maleimide
11 polyethylene glycol biotin ester of amine.
(5) reaction solution in (4) is adjusted to 1mg/mL using the PBS solution of PH=7.4.
(6) CEA, NSE, ProGRP are modified respectively using above-mentioned biotin modification method, SCC, CYFRA21-1 label are anti-
Body.
The step of embodiment 3 is using magnetosensitive immune detection card of the present invention detection five tumor markers of lung cancer
(1) it prepares and contains 50 μ g/mL CEA, 50 μ g/mL NSE, 50 μ g/mL ProGRP, 50 μ g/mL SCC, 50 μ g/mL
The mixed solution of the biotinylated labelled antibody of CYFRA21-1.
(2) 75mg/mL Streptavidin magnetic particle solution is prepared.
The mixed solution of (3) 5 μ L biotinylation labelled antibodies and 5 μ L Streptavidin magnetic particle solutions are using freeze-drying
Technique be separately stored in labelled antibody and magnetic particle freeze-drying slot in.
(4) reagent storage layer cleaning liquid pool and magnetic grain dissolution liquid pool store 100 μ L cleaning solutions and 100 μ L magnetic particles respectively
Redissolve liquid.
(5) the PDMS sealing cover above sample cell is opened, 50 μ L sample solutions are added in sample cell.
(6) the PDMS sealing cover above sample cell is pressed, sample solution is injected into sample level sensing position, then,
The PDMS sealing cover above pressing sample cell shakes sample solution repeatedly in labelled antibody freeze-drying area up and down, it is ensured that labelled antibody
It is completely dissolved in sample solution.
(7) continue to press PDMS sealing cover, labelled antibody is made to be injected into GMR chip list together with the mixed solution of sample
Face, by antigen in coated antibody-sample-labelled antibody sandwich immunoassay reaction pattern, the labelled antibody of biotin modification is fixed
In chip surface.
(8) PDMS sealing cover above clear particle is pressed, cleaning solution is injected, extra antigen is washed away with labelled antibody.
(9) pressing magnetic particle answers superjacent PDMS sealing cover, and magnetic particle redissolution liquid is injected into magnetic particle level sensing
Position, shaking magnetic particle redissolution liquid repeatedly is completely dissolved the magnetic particle of freeze-drying,.
(10) continue pressing magnetic particle and answer superjacent PDMS sealing cover, magnetic particle solution is injected into chip surface, is led to
Biotin-avidin system is crossed, the magnetic particle of Streptavidin modification is captured to chip surface.In the influence of external magnetic field
Under, magnetic particle generates derivative magnetic field, causes GMR reaction zone resistance variations.
The case where magnetic particle is captured to chip surface is as shown in Figure 3；Five tumor-markers of lung cancer in embodiment 3
The microchannel structure of the magnetosensitive immune detection card of object is as shown in Figure 4.
4 lung cancer of embodiment, five tumor markers magnetosensitive immune detection cards are to CEA, NSE, ProGRP, SCC, CYFRA21-1
Joint-detection
Configuration is carried out according to the following table in antigenic solution, configures 8 sample solutions, CEA, NSE, SCC, CYFRA21-1 concentration altogether
It is ng/mL, ProGRP concentration is pg/mL.
8 sample solutions are successively tested, corresponding magnetic signal value is obtained, according to the relationship of antigen concentration and magnetic signal value,
Draw corresponding standard curve, such as Fig. 5, the logarithm of magnetic signal value and analyte concentration shows good linear relationship.
Obviously, the above embodiment of the present invention be only to clearly illustrate example of the present invention, and not be pair
The restriction of embodiments of the present invention may be used also on the basis of the above description for those of ordinary skill in the art
To make other variations or changes in different ways, all embodiments can not be exhaustive here, it is all to belong to this hair
The obvious changes or variations that bright technical solution is extended out are still in the scope of protection of the present invention.
Claims (10)
1. a kind of GMR chip, which is characterized in that the surface of the GMR chip includes poly- for capturing the maleic anhydride of antibody
Close object film layer.
2. GMR chip according to claim 1, which is characterized in that the maleic anhydride polymer film layer with a thickness of
6-42nm。
3. according to claim 1 or 2 described in any item GMR chips, which is characterized in that the maleic anhydride polymer film layer
It is modified by the method for spin coating maleic anhydride polymer solution in chip surface.
4. GMR chip according to claim 3, which is characterized in that the process of spin coating includes: 500-1500 revs/min of spin coating
6s；Then 5000-10000 revs/min of spin coating 20s.
5. according to claim 1 or 2 described in any item GMR chips, which is characterized in that the maleic anhydride polymer is selected from poly-
Thylene maleic acid anhydride copolymer, polystyrene maleic anhydride copolymer, polystyrene graft maleic anhydride, polycthylene grafted Malaysia
One or more of acid anhydrides, polypropylene grafted maleic anhydride and polyisoprene grafted maleic anhydride.
6. according to claim 1 or 2 described in any item GMR chips, which is characterized in that the maleic anhydride polymer is polyphenyl
Vinyl Graft maleic anhydride.
7. according to claim 1 or 2 described in any item GMR chips, which is characterized in that if the GMR chip includes dry reaction
Area, each reaction zone can a kind of albumen of independent detection.
8. such as application of the described in any item GMR chips of claim 1-7 in polyprotein joint-detection.
9. a kind of magnetosensitive immune detection card for Protein Detection, which is characterized in that including as described in claim any one of 1-7
GMR chip.
10. magnetosensitive immune detection card according to claim 9, which is characterized in that from top to bottom successively include top cover, PDMS
Sealing cover, reagent storage layer, PDMS sealing film, printed circuit board and bottom cover；The GMR chip is pasted onto the printed circuit board
Above and with printed circuit board it is electrically connected.
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