CN107091821A - The preparation method on the surface plasma resonance instrument chip antipollution surface modified based on mucoprotein - Google Patents
The preparation method on the surface plasma resonance instrument chip antipollution surface modified based on mucoprotein Download PDFInfo
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- CN107091821A CN107091821A CN201710209595.5A CN201710209595A CN107091821A CN 107091821 A CN107091821 A CN 107091821A CN 201710209595 A CN201710209595 A CN 201710209595A CN 107091821 A CN107091821 A CN 107091821A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/55—Specular reflectivity
- G01N21/552—Attenuated total reflection
- G01N21/553—Attenuated total reflection and using surface plasmons
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Abstract
The invention discloses a kind of preparation method on the surface plasma resonance instrument chip antipollution surface modified based on mucoprotein, amphipathic mucoprotein is connected to golden film surface by the hydrophobic grouping of peptide backbone, self assembled monolayer is formed, so as to obtain a kind of surface plasma resonance instrument chip based on amphipathic mucoprotein.The chip of the present invention is respectively provided with excellent antifouling property in the complex system of single albumen system and pH=7.4, and is easy to self-assemble to various surfaces, and regular monolayer is stablized in formation.The features such as chip of the present invention has excellent antifouling property and high stability, and preparation method is easy, with good feasibility and universality.
Description
Technical field
It is more particularly to a kind of based on viscous egg the present invention relates to surface plasma resonance spectrometer antipollution chip preparation field
The preparation method on the surface plasma resonance instrument chip antipollution surface modified in vain.
Background technology
Surface plasma resonance (Surface Plasmon Resonance, abbreviation SPR) instrument is that occur 1980s
A kind of biosensor technology, when incident light incides low refractive index dielectric from high refractive index medium, it may occur that total reflection,
If now plating layer of metal film (gold or silver) at dielectric interface, the evanescent wave that incident light is produced can cause metal surface certainly
Occur collective oscillation by electronics, and then form plasma, when evanescent wave and the frequency and wave number phase of surface plasmon oscillations
Deng when, that is, produce covibration (SPR), cause energy from evanescent wave is transferred to surface plasma-wave, make the intensity of reflected light
Weaken significantly, decay total reflection phenomenon, the incidence angle referred to as resonance angle when the intensity of reflected light is zero is presented.Resonance angle and gold
Belong to film interface medium refraction index relevant, and refractive index changes with combination in the molecular mass of metal film surfaces, therefore can
The information of intermolecular interaction is obtained by analyzing resonance angle, because it has detection quick and the features such as need not mark,
It has been widely used in the fields such as proteomics, medicament research and development, clinical diagnosis, food security and environmental monitoring, and has shown
Go out wide application prospect.
SPR chips be surface plasma resonance instrument core the most component there is provided produce spr signal necessary physics bar
Part, mainly including coupled apparatus, metal film and surface matrix.However, when carrying out analysis test using surface plasma resonance instrument,
The surface contamination of sensing chip is a common problem, the non-specificity of biomolecule or microorganism in sample
The accuracy quantitatively detected will be reduced by adsorbing, and produce false positive results.Therefore, exploitation is effective against the surface of non-specific adsorption
It is the matter of utmost importance for improving surface plasma resonance sensor sensitivity and accuracy.
The content of the invention
For the deficiencies in the prior art, the present invention provides a kind of surface plasma resonance modified based on mucoprotein
The preparation method on instrument chip antipollution surface.
The present invention is to be realized by the following technical programs：
A kind of preparation method on the surface plasma resonance instrument chip antipollution surface modified based on mucoprotein, its feature is existed
In comprising the following steps：
A) prepared by naked golden chip
The thick layers of chrome of one layer of 2-3nm is coated in substrate using electron beam evaporation deposition technology, one is coated in the layers of chrome
Golden film thick layer 45-50nm, obtains naked golden chip；
B) naked golden chip pretreatment
Step a) the naked golden chips obtained are immersed in the mixed solution of water, 30% concentrated ammonia liquor and 30% hydrogen peroxide, it is above-mentioned
The volume ratio of mixed solution reclaimed water, 30wt.% concentrated ammonia liquors and 30wt.% hydrogen peroxide is 3:1:1,7 at 70-90 DEG C in soaking 10-
60min, taking-up is cleaned after 3 times with deionized water and dried up with nitrogen；
C) mucoprotein solution is pre-processed
Prepare 2-10mg/mL mucoprotein solution with pH=7.4 phosphate buffer, by the albumen assembled in above-mentioned solution with
9000-12000rpm centrifugations 10-20min is removed；
D) mucoprotein is modified
Fitted to after the chip cleaning that b) step is obtained with pitch on the prism of surface plasma resonance instrument system, with pH
It is mobile phase for 7.4 phosphate buffer, flow velocity is that 50-100 μ L/min flow continuously through chip surface, when baseline stability, note
Enter the pretreated mucoproteins of 100-200 μ L, and flow cell is flowed through with 20-50 μ L/min speed, surface plasma occur and be total to
During vibration response signal, stop flow velocity, be incubated 10-20min, then buffer solution 10-30min is walked with 50-100 μ L/min flow velocity, clearly
Wash uncombined firm mucoprotein off.
Moreover, above-mentioned substrate is glass chip bottom or optical fiber substrate, the preferred BK7 substrate of glass in glass chip bottom.
Moreover, above-mentioned golden film can also be silverskin or silane film or silver/gold composite membrane.
Moreover, above-mentioned c) mucoprotein described in step secreted from the stomach of pig.
Advantages of the present invention and beneficial effect are：
(1) surface plasma resonance instrument chip modified based on mucoprotein prepared by the present invention has excellent resistance tocrocking
Energy；
(2) present invention prepare based on mucoprotein modify surface plasma resonance instrument chip have good lubricity and
Bio-compatibility；
(3) surface plasma resonance instrument chip modified based on mucoprotein prepared by the present invention has high stability, dries
After storage being stored under 2 months or room temperature condition under state 3 months, antifouling property is unchanged；
(4) preparation method for the surface plasma resonance instrument chip that the present invention is modified based on mucoprotein is easy to be quick, reduces
Conventional multistep cumbersome preparation process, drastically increases the efficiency of chip surface modification；
(5) the antipollution material tool on the surface plasma resonance instrument chip surface modified based on mucoprotein prepared by the present invention
There is biodegradability.
Brief description of the drawings
The structural representation for the surface plasma resonance instrument chip that Fig. 1 is modified for the present invention based on mucoprotein；
Fig. 2 is surface plasma resonance instrument chip of the present invention based on mucoprotein modification to 2mg/mL lysozymes
(lysozyme), in 2mg/mL bovine serum albumins (BSA) and 0.5mg/mL sheep immunoglobulin (IgG) solution albumen it is non-
Specific adsorption amount；
Wherein, 1, glass chip bottom；2nd, layers of chrome；3rd, golden membranous layer；4th, mucoprotein self assembled monolayer.
Embodiment
The present invention is described in further detail by following examples.It should be noted that：Following embodiments are illustrative, are not
Limited, it is impossible to limit protection scope of the present invention with following embodiments.
Embodiment 1
The thick layers of chrome of one layer of 2nm is coated in BK7 substrate of glass using electron beam evaporation deposition technology, is applied in the layers of chrome
The thick golden films of one layer of 48nm are covered, naked golden chip is obtained；By above-mentioned naked golden chip immersion water, 30% concentrated ammonia liquor and 30% hydrogen peroxide
In mixed solution, the volume ratio of water, 30% concentrated ammonia liquor and 30% hydrogen peroxide is 3:1:1, in soaking 40min at 70 DEG C.Then take
Go out and cleaned with deionized water 3 times, dried up with nitrogen.2mg/mL is prepared with pH=7.4 phosphate buffer to secrete from the stomach of pig
The mucoprotein solution obtained, the albumen assembled in above-mentioned solution is removed with 9000rpm centrifugations 10min；The core after drying will be cleaned
Piece is fitted to pitch on the prism of surface plasma resonance system, using pH be 7.4 phosphate buffer as mobile phase, selection stream
Speed flows continuously through chip surface for 50 μ L/min, when baseline stability, injects the 100 pretreated mucoproteins of μ L, and with 20 μ L/
Min speed flows through flow cell, when there is surface plasma resonance response signal, stops flow velocity, is incubated 10min, then with 50 μ L/
Min flow velocity walks buffer solution 20min, washes uncombined firm mucoprotein.It is flowing by 7.4 phosphate buffer of pH
Phase, selection flow velocity is 50 μ L/min.After baseline is steady, 100 μ L protein solution is injected separately into quantitative loop：0.5mg/mL
Sheep immunoglobulin, 2mg/mL bovine serum albumin(BSA) and 2mg/mL lysozyme, promote above-mentioned each albumen molten through mobile phase
Liquid reaches chip surface, read after surface plasma resonance system measurement resonance angle real-time change curve, 20min surface etc. from
Sub-resonance angle changing value, and calculate non-specific adsorption amount respectively 18.16ng/cm2、26.15ng/cm2、10.92ng/cm2。
Embodiment 2
The thick layers of chrome of one layer of 3nm is coated in BK7 substrate of glass using electron beam evaporation deposition technology, is applied in the layers of chrome
The thick golden films of one layer of 47nm are covered, naked golden chip is obtained；By above-mentioned naked golden chip immersion water, 30% concentrated ammonia liquor and 30% hydrogen peroxide
In mixed solution, the volume ratio of water, 30% concentrated ammonia liquor and 30% hydrogen peroxide is 3:1:1, in soaking 15min at 85 DEG C.Then take
Go out and cleaned with deionized water 3 times, dried up with nitrogen.10mg/mL is prepared with pH=7.4 phosphate buffer to secrete from the stomach of pig
The mucoprotein solution obtained, the albumen assembled in above-mentioned solution is removed with 12000rpm centrifugations 10min；The core after drying will be cleaned
Piece is fitted to pitch on the prism of surface plasma resonance system, using pH be 7.4 phosphate buffer as mobile phase, selection stream
Speed flows continuously through chip surface for 100 μ L/min, when baseline stability, injects the 200 pretreated mucoproteins of μ L, and with 50 μ
L/min speed flows through flow cell, when there is surface plasma resonance response signal, stops flow velocity, is incubated 10min, then with 100
μ L/min flow velocity walks buffer solution 10min, washes uncombined firm mucoprotein.It is stream by 7.4 phosphate buffer of pH
Dynamic phase, selection flow velocity is 50 μ L/min.After baseline is steady, 100 μ L protein solution is injected separately into quantitative loop：0.5mg/
The lysozyme of mL sheep immunoglobulin, 2mg/mL bovine serum albumin(BSA) and 2mg/mL, above-mentioned each albumen is promoted through mobile phase
Solution reaches chip surface, and surface etc. is read after surface plasma resonance system measurement resonance angle real-time change curve, 20min
Ion resonance angle changing value, and calculate non-specific adsorption amount.
The surface plasma resonance instrument chip modified based on mucoprotein prepared by the present invention has excellent antifouling property,
75% is reduced to the non-specific adsorption amount of lysozyme, bovine serum albumin, sheep immunoglobulin solution albumen compared to naked gold
More than；With good lubricity and bio-compatibility；With high stability, 2 months or room temperature condition are stored under drying regime
After lower storage 3 months, antifouling property is unchanged；The antipollution material of chip surface has biodegradability；And this hair
Bright preparation method is easy to be quick, reduces the cumbersome preparation process of conventional multistep, drastically increases chip surface modification
Efficiency.
Exemplary description is done to the present invention above, it should explanation, in the situation for the core for not departing from the present invention
Under, any simple deformation, modification or other skilled in the art can not spend the equivalent substitution of creative work equal
Fall into protection scope of the present invention.
Claims (4)
1. a kind of preparation method on the surface plasma resonance instrument chip antipollution surface modified based on mucoprotein, its feature is existed
In comprising the following steps：
A) prepared by naked golden chip
The thick layers of chrome of one layer of 2-3nm is coated in substrate using electron beam evaporation deposition technology, one layer of 45- is coated in the layers of chrome
Golden film thick 50nm, obtains naked golden chip；
B) naked golden chip pretreatment
Step a) the naked golden chips obtained are immersed in the mixed solution of water, 30wt.% concentrated ammonia liquors and 30wt.% hydrogen peroxide, on
The volume ratio for stating mixed solution reclaimed water, 30wt.% concentrated ammonia liquors and 30wt.% hydrogen peroxide is 3:1:1, in soaking 10- at 70-90 DEG C
60min, taking-up is cleaned after 3 times with deionized water and dried up with nitrogen；
C) mucoprotein solution is pre-processed
Prepare 2-10mg/mL mucoprotein solution with pH=7.4 phosphate buffer, by the albumen assembled in above-mentioned solution with
9000-12000rpm centrifugations 10-20min is removed；
D) mucoprotein is modified
Fitted to after the chip cleaning that b) step is obtained with pitch on the prism of surface plasma resonance instrument system, using pH as
7.4 phosphate buffer is mobile phase, and flow velocity is that 50-100 μ L/min flow continuously through chip surface, when baseline stability, injection
The pretreated mucoproteins of 100-200 μ L, and flow cell is flowed through with 20-50 μ L/min speed, there is surface plasma resonance
During response signal, stop flow velocity, be incubated 10-20min, then buffer solution 10-30min is walked with 50-100 μ L/min flow velocity, clean
Fall uncombined firm mucoprotein.
2. preparation method according to claim 1, it is characterised in that described substrate is glass chip bottom or optical fiber base
Bottom.
3. preparation method according to claim 1, it is characterised in that described golden film can also for silverskin or silane film or
Silver/golden composite membrane.
4. preparation method according to claim 1, it is characterised in that c) mucoprotein described in step is the stomach from pig
Secrete.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201710209595.5A CN107091821A (en) | 2017-03-31 | 2017-03-31 | The preparation method on the surface plasma resonance instrument chip antipollution surface modified based on mucoprotein |
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| CN201710209595.5A CN107091821A (en) | 2017-03-31 | 2017-03-31 | The preparation method on the surface plasma resonance instrument chip antipollution surface modified based on mucoprotein |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108931647A (en) * | 2018-07-06 | 2018-12-04 | 深圳信息职业技术学院 | The production method of Fiber imunosensor, detection device and Fiber imunosensor |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101477114A (en) * | 2009-01-22 | 2009-07-08 | 中国医学科学院生物医学工程研究所 | Production method of surface plasma resonance chip |
| CN105548085A (en) * | 2015-12-04 | 2016-05-04 | 天津大学 | Preparation method of surface plasma resonance instrument chip based on amphiphilic heptapeptide modification |
-
2017
- 2017-03-31 CN CN201710209595.5A patent/CN107091821A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101477114A (en) * | 2009-01-22 | 2009-07-08 | 中国医学科学院生物医学工程研究所 | Production method of surface plasma resonance chip |
| CN105548085A (en) * | 2015-12-04 | 2016-05-04 | 天津大学 | Preparation method of surface plasma resonance instrument chip based on amphiphilic heptapeptide modification |
Non-Patent Citations (2)
| Title |
|---|
| DEBBY P. CHANG,ET.AL: "Conformational Mechanics, Adsorption, and Normal Force Interactions of Lubricin and Hyaluronic Acid on Model Surfaces", 《LANGMUIR》 * |
| 赵策洲 等: "《半导体硅基材料及其光波导》", 31 August 1997 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108931647A (en) * | 2018-07-06 | 2018-12-04 | 深圳信息职业技术学院 | The production method of Fiber imunosensor, detection device and Fiber imunosensor |
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Application publication date: 20170825 |