CN107632151B - Solid phase material for biological detection and analysis - Google Patents
Solid phase material for biological detection and analysis Download PDFInfo
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- CN107632151B CN107632151B CN201710631319.8A CN201710631319A CN107632151B CN 107632151 B CN107632151 B CN 107632151B CN 201710631319 A CN201710631319 A CN 201710631319A CN 107632151 B CN107632151 B CN 107632151B
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Abstract
The invention discloses a solid phase material for biological detection and analysis. The material is characterized in that: microspheres are fixed on the detection surface layer of the microporous plate (or the card strips forming the microporous plate), and the material can effectively improve the binding quantity among biological ligand molecules, so that the detection sensitivity is improved, such as: specific antigen-antibody, biotin-avidin, enzyme-substrate, Aptamer (Aptamer) -protein, nucleic acid complementary base pair; can be applied to immunology, biochemistry or molecular biology. The material can replace the prior conventional enzyme-linked immunosorbent and chemiluminescent microporous plate.
Description
Technical field
The present invention relates to a kind of for Biological Detection, the solid phase material of analysis, belong to biology, agronomy, zoology and
The technical field of medical research, inspection, detection.
Background technique
Biology, agronomy, zoology and medical research, inspection, detection many applications in using microwell plate (or group
At the strip of microwell plate, Fig. 1 and Fig. 2 are seen).Microplate is usually by the polymer material of the shape of a matrix.Microwell plate (or
Form the strip of microwell plate) it is generally injection molding.Microwell plate it is relatively common have 96,386 hole equal-specifications, have regularly arranged
Same unit lattice.Generally each cell is a hole, receives, storage, includes liquid, fixing biological molecules and its guarantee are special
Fixed reaction carries out.With the development of industry, also there are more and more strips as independent detection unit.Strip is general
For 8 holes or 12 holes, calibration, Quality Control and detection sample can be realized in an independent strip detection unit.Nowadays, relevant
Biochemical Research is growing, has been advised greatly using microwell plate as the protein of carrier, nucleic acid and other biological crystal product
Mould industrialized production is not limited only to the research of academia.
Ultra-violet curing is a kind of using the curing materials containing photoinitiator, generates living radical under ultraviolet light
Or cation, cause monomer polymerization, be crosslinked and connect branch chemical reaction, it is made to be converted into solid-state by liquid.Using the property, select
It is as connecting material.With the development of science and technology, the product of the every field such as electronics, medical treatment rapidly develops, it is viscous to ultra-violet curing
The demand of knot mode is increasing.Ultraviolet light belongs to black light, is one section of electromagnetic radiation other than visible light, wavelength is in 10-
The range of 400nm.Ultra-violet curing principle is: the photoinitiator in ultraviolet-curable materials absorbs ultraviolet under ultraviolet irradiation
Generate living radical or cation after light, cause monomer polymerization, crosslinking and section branch chemical reaction, make adhesive in a few seconds by
Liquid is converted into solid-state.Ultra-violet curing has begun working on and applies very early, and last century 60, the seventies have had a lot special
Benefit occurs, and just mentioning in U.S.Pat.No.3,699,089 can star with ultraviolet light or accelerate polymerization reaction,
U.S.Pat.No.3 has been defined in 864,143 and has been applied to the modification of ethylene film surface in ultra-violet curing, U.S.Pat.No.4,
Ultra-violet curing is applied to the method that a manufacture has the plastic lens original part of graded index by 022,855 this invention.
For microballoon as solid phase, combined surface area is big, and surface is easy to modify chemical group and for being total to biomolecule
Valence combines, and controllably, still, microballoon itself is not easy to be fixed the size of microballoon as solid phase carrier, gives biomolecule
Cleaning step in reaction process brings difficulty, and cleaning step could be completed by needing specific process and additional equipment to cooperate,
Increase technical difficulty and cost.Although conventional microwell plate is readily cleaned, but bonded area is limited, the knot with biomolecule
It closes and leans on passive adsorption.In the patent U.S.Pat.No.4 of early stage, 205,954, has begun and microballoon is applied to immune detection
In reaction, for promoting the detection performance of immunoturbidimetry detection method.
Microballoon refers to skin effect: the ratio between the surface atom number of ultrafine dust and total atom number are as diameter of particle becomes
It is small and increased dramatically, it the crystalline field environment of surface atom and combines and can have very big chemical activity with the difference of interior atoms,
Its surface can greatly increase.This is because lack adjacent atom around surface atom and have unsaturation, easily with other originals
Son combine and settle out, it is seen that skin effect be it is a kind of influence chemical characteristic factor.
Meanwhile also there is microballoon bulk effect to refer to that the atomicity for including due to ultrafine dust is reduced and adds band power levels
Greatly, so that some physical properties of substance be made to be abnormal because of the discontinuity of energy level.
The two effects of microballoon can specifically be reacted increases sharply in microballoon specific surface area, and microballoon functional group densities and selectivity are inhaled
Receipts ability becomes larger, and the time for reaching adsorption equilibrium shortens, and the stability of particle greatly improves.
Therefore, it is developed based on the easily operated of microwell plate, while the efficient advantage of microballoon can be taken into account, for biology
The solid phase material examined and analyzed is learned, will be highly beneficial.
Summary of the invention
Technical problem: as described above in order to overcome, microballoon is not easy to be fixed as solid phase, and cleaning is difficult, increases skill
Disadvantage and microwell plate (or strip of composition microwell plate) bonded area of art and cost is limited, passive adsorption biomolecule
It is insufficient.The present invention provides one kind can take into account microballoon biomolecular binding abilities and microwell plate (or strip of composition microwell plate)
The solid phase material of easy cleaned advantage.
Technical solution: the purpose of the present invention can be achieved through the following technical solutions.
Step 1: by being cleaned and dried to obtain clean detection surface, this detection surface specifically: microwell plate (or composition
The strip of microwell plate) internal surface of hole, the material on surface can be specially polystyrene, polytetrafluoroethylene PTFE, EU, polyvinyl chloride
One of PVC, polypropylene, polyethylene, silica etc., it is most common be polystyrene, using acid, ethyl alcohol, purified water according to
Secondary cleaning.
Step 2: preparing photocuring reaction liquid, and n-isopropyl acrylamide, acrylic acid are dissolved in dimethyl sulfoxide, added
Enter anthraquinone-2-sodium, adds isopropanol.
Step 3: microballoon is added in above-mentioned reaction solution, is uniformly mixed, final concentration of the microballoon in reaction solution is in 0.1-
Within the scope of 5mg/ml.
Step 4: the horizontal rest, to height on clean detection surface is added dropwise in the above-mentioned reaction solution containing microballoon
Pressure mercury lamp irradiation, the wavelength of irradiation are 300-400nm, and irradiation time is 5-30 minutes.
Step 5: after illumination, surface is successively detected with dehydrated alcohol, distilled water flushing, to remove unreacted reactant, is done
The microwell plate that microballoon layer is distributed with is obtained after dry (wherein a certain single hole schematic diagram is shown in Fig. 3).
The partial size of above-mentioned microballoon is within the scope of 0.1-5um.
Above-mentioned microballoon can be the bare ball not combined in advance with biomolecule, be also possible to tie in advance with biomolecule
Label ball after conjunction.
The bare ball surface of above-mentioned microballoon can be modified without chemical group, can also modify in advance amino, carboxyl, hydroxyl,
The chemical groups such as aldehyde radical.
The novel microporous plate obtained through the above technical solution, more common microwell plate are considerably increased for biomolecule knot
The surface area of conjunction can be improved 2-10 times, and then improve the reaction efficiency of biomolecule, enhance application performance.
Detailed description of the invention
96 hole microwell plate schematic diagram (top view) of Fig. 1.
8 hole strip schematic diagram of Fig. 2 (being from top to bottom respectively top view, side view, lateral sectional view)
Fig. 3 single hole schematic diagram (is respectively novel microporous plate single hole sectional view, the conventional microporous after microballoon combines from top to bottom
Plate single hole sectional view)
Specific embodiment
Below with reference to embodiment, the present invention is described in more detail, but the present invention is not limited only to these embodiments.
Embodiment one: labelled AFP (α-fetoprotein, AFP) antibody is used for after microballoon and detection surface cure
Detection)
1. using 0.32cm2Microwell plate (or the composition microwell plate of hole floor space (i.e. common 96 hole sizes) polystyrene
Strip) internal surface of hole cleaned as detection surface using the cleaning way of above-mentioned steps one, and it is dry.Microwell plate is purchased from upper
Extra large brilliant peaceful object, article No. J09604.
2. preparing photocuring reaction liquid, specific ratio are as follows: n-isopropyl acrylamide using the mode of above-mentioned steps two
10%, acrylic acid 8% is dissolved in dimethyl sulfoxide, is added 0.6% anthraquinone-2-sodium, then plus 0.3% enters isopropanol.
3. using polystyrene microsphere, partial size 1um, microsphere surface has modified carboxylic group in advance.It, will according to step 3
Microballoon is added in photocuring reaction liquid, and the ultimate density of microballoon is 1mg/ml.Microballoon is purchased from Beijing Hua Taixin biologic medical skill
Art Co., Ltd, brand Spherotech, article No. CP-10-100.
4. above-mentioned microballoon reaction solution is added on micropore detection surface, additional amount is the hole 75ul/, so according to step 4
It is placed on shaker (company, celo Czech, the U.S., Beijing Ke Bosaier Biotechnology Co., Ltd agency), shaken at room temperature 15
Minute, vibration velocity 350, until microballoon is uniformly paved with bottom hole；Ultraviolet curing device select Hebei Zhuozhou hundred think scientific and technological 102 type UV is solid
Change machine: irradiation condition are as follows: wavelength 365nm, irradiation time are 20 minutes.
5. completing cleaning according to step 5, novel microporous plate (or strip of composition microwell plate) material is obtained.
6. 1- (3- the dimethylamino-propyl) -3- ethyl carbodiimide and 90ul concentration that are successively 2mg/ml by 10ul concentration
It is added in new material hole for alpha-fetoprotein (AFP) antibody of 1ug/ml, after room temperature 1 reacts hour, after dry after cleaning, is used for
The micropore board detecting method of alpha-fetoprotein.1- (3- dimethylamino-propyl) -3- ethyl carbodiimide is purchased from Sigma-Aldrich,
Article No. E6383.Antibody is purchased from U.S. Meridian Life Science company, article No. H45610M.
7. by novel alpha-fetoprotein (AFP) microwell plate, another AFP antibody of horseradish peroxidase-labeled, then cooperate with
Cleaning solution, chemiluminescent substrate form detection reagent, detect a series of AFP antigen of various concentrations.It is coated with traditional approach
AFP antibody microwell plate compares, and the conditions such as other marker concentrations, reaction time, cleaning process are consistent.Horseradish peroxidating
Object enzyme is purchased from Sigma-Aldrich, article No. P8375.Label is purchased from U.S. Meridian Life Science company with antibody,
Article No. H45301M.Chemiluminescence detector device is purchased from Beijing Pu Lang Technew SA, instrument model PHX-2012.It washes
Trigger is purchased from Beijing Pu Lang Technew SA, instrument model DNX-9620.Chemiluminescent substrate is purchased from the scorching prosperous life in Shanghai
Object Science and Technology Ltd., article No. ECL-P-100.From testing result it can be seen that the detection performance of novel microporous plate is substantially better than
Conventional microplates.
Detection data under above-mentioned condition is as follows:
Data in table 1 can be seen that novel microporous plate measured value more luminous than the bulk chemical of conventional microplates is considerably higher,
The association reaction of more antigens and antibody has occurred on novel microporous plate.
Data in table 2 can be seen that increasing with concentration of specimens, the increasing degree of the relative deviation of conventional microplates
It is apparently higher than novel microporous plate, the accurate detection range of novel microporous plate is wider than conventional microplates.Concentration of specimens in table is warp
Alpha-fetoprotein antigen concentration in the human serum that commercially available reagent detects.
It is in table 3 statistics indicate that the accuracy of novel microporous plate is substantially better than conventional microporous when detecting low concentration sample
Plate, i.e., novel microporous plate can be such that the detection sensitivity of reagent obviously optimizes.
Embodiment two: solidify again through ultraviolet light for detecting cardiac muscle troponin I after microballoon and Streptavidin coupling
The detection of (cardiac Troponin I, cTnI)
1. using polystyrene microsphere, partial size 1um, microsphere surface has modified carboxylic group in advance, by microballoon and strepto-
Avidin, 1- (3- dimethylamino-propyl) -3- ethyl carbodiimide are uniformly mixed, and the final concentration of 10mg/ml of microballoon, strepto- is affine
Final concentration of 1mg/ml, the final concentration of 1mg/ml of 1- (3- dimethylamino-propyl) -3- ethyl carbodiimide of element, room temperature reaction
It is stand-by after cleaning after 2 hours.1- (3- dimethylamino-propyl) -3- ethyl carbodiimide is purchased from Sigma-Aldrich, article No.
E6383.Streptavidin is purchased from Sigma-Aldrich, article No. 85878.Microballoon has purchased from Beijing Hua Taixin bio-medical technology
Limit company, brand Spherotech, article No. CP-10-100.
2. using 0.32cm2Polystyrene micropore plate (or the composition of the hole floor space floor spaces of 96 hole sizes (i.e. common)
The strip of microwell plate) micropore inner surface cleaned as detection surface using the cleaning way of above-mentioned steps one, and it is dry.It is micro-
Orifice plate is purchased from the brilliant peaceful object in Shanghai, article No. J09604.
3. preparing photocuring reaction liquid, specific ratio are as follows: n-isopropyl acrylamide using the mode of above-mentioned steps two
10%, acrylic acid 8% is dissolved in dimethyl sulfoxide, is added 0.6% anthraquinone-2-sodium, then plus 0.3% enters isopropanol.
4. the microballoon for having marked Streptavidin is added in photocuring reaction liquid according to step 3, microballoon it is final
Concentration is 1mg/ml.
5. above-mentioned microballoon reaction solution is added on micropore detection surface, additional amount is the hole 75ul/, so according to step 4
It is placed on shaker (company, celo Czech, the U.S., Beijing Ke Bosaier Biotechnology Co., Ltd agency), shaken at room temperature 15
Minute, vibration velocity 350, until microballoon is uniformly paved with bottom hole；Ultraviolet curing device select Hebei Zhuozhou hundred think scientific and technological 102 type UV is solid
Change machine: irradiation condition are as follows: wavelength 365nm, irradiation time are 20 minutes.
6. according to step 5, cleaning is completed, obtains novel microporous plate (or the card of composition microwell plate of marked by streptavidin
Item) material.For microwell plate biotin-avidin detection system.
By novel Streptavidin microwell plate, cTnI antibody A with biotin labeling, horseradish peroxidase-labeled
CTnI antibody B, then cooperate with cleaning solution, chemiluminescent substrate, detection reagent is formed, a series of cTnI for detecting various concentrations is anti-
It is former.It is compared with the coated Streptavidin microwell plate of traditional approach, other marker concentrations, reaction time, cleaning process
Etc. conditions it is consistent.Biotin is purchased from Sigma-Aldrich, article No. B4639.Antibody A is public purchased from Finland Hytest biotechnology
Department, article No. 4T21-560.Antibody B is purchased from Finland Hytest biotechnology company, article No. 4T21-458.Horseradish peroxidase
Enzyme is purchased from Sigma-Aldrich, article No. P8375.Chemiluminescence detector device is purchased from Beijing Pu Lang Technew SA, instrument
Model PHX-2012.Board-washing machine is purchased from Beijing Pu Lang Technew SA, instrument model DNX-9620.Chemiluminescence bottom
Object is purchased from Shanghai Yan Xi Biotechnology Co., Ltd, article No. ECL-P-100.From testing result it can be seen that novel microporous plate
Detection performance is substantially better than conventional microplates.
Detection data under above-mentioned condition is as follows:
Data in table 4 can be seen that novel microporous plate measured value more luminous than the bulk chemical of conventional microplates is considerably higher,
The association reaction of more antigens and antibody has occurred on novel microporous plate.
Data in table 5 can be seen that increasing with concentration of specimens, the increasing degree of the relative deviation of conventional microplates
It is apparently higher than novel microporous plate, the accurate detection range of novel microporous plate is wider than conventional microplates.Concentration of specimens in table is warp
Alpha-fetoprotein antigen concentration in the human serum that commercially available reagent detects.
It is in table 6 statistics indicate that the accuracy of novel microporous plate is substantially better than conventional microporous when detecting low concentration sample
Plate, i.e., novel microporous plate can be such that the detection sensitivity of reagent obviously optimizes.
Embodiment three: microballoon and detection surface cure after be coated with carcinomebryonic antigen (carcino-embryonicantigen,
CEA) antibody is for detecting
1. using 0.32cm2Microwell plate (or the composition microwell plate of hole floor space (i.e. common 96 hole sizes) polystyrene
Strip) internal surface of hole cleaned as detection surface using the cleaning way of above-mentioned steps one, and it is dry.Microwell plate is purchased from upper
Extra large brilliant peaceful object, article No. J09604.
2. preparing photocuring reaction liquid, specific ratio are as follows: n-isopropyl acrylamide using the mode of above-mentioned steps two
10%, acrylic acid 8% is dissolved in dimethyl sulfoxide, is added 0.6% anthraquinone-2-sodium, then plus 0.3% enters isopropanol.
3. using polystyrene microsphere, partial size 1um, microsphere surface has modified carboxylic group in advance.It, will according to step 3
Microballoon is added in photocuring reaction liquid, and the ultimate density of microballoon is 1mg/ml.Microballoon is purchased from Beijing Hua Taixin biologic medical skill
Art Co., Ltd, brand Spherotech, article No. CP-10-100.
4. above-mentioned microballoon reaction solution is added on micropore detection surface, additional amount is the hole 75ul/, so according to step 4
It is placed on shaker (celo Czech, the U.S., Beijing Ke Bosaier Biotechnology Co., Ltd agency), shaken at room temperature 15 minutes,
Vibration velocity 350, until microballoon is uniformly paved with bottom hole；Ultraviolet curing device selects Hebei Zhuozhou hundred to think to obtain scientific and technological 102 type UV curing:
Irradiation condition are as follows: wavelength 365nm, irradiation time are 20 minutes.
5. completing cleaning according to step 5, novel microporous plate (or strip of composition microwell plate) material is obtained.
6. the CEA antibody that 100ul concentration is 1ug/ml is added in new material hole, it is placed on 2-8 DEG C and is coated with 14 hours
Afterwards, after dry after cleaning, the micropore board detecting method for CEA.Antibody is public purchased from U.S. Meridian Life Science
Department, article No. M01250M.
7. by novel C EA microwell plate, another CEA antibody of horseradish peroxidase-labeled, then cooperate with cleaning solution, chemistry
Luminous substrate forms detection reagent, detects a series of CEA antigen of various concentrations.With the coated CEA antibody micropore of traditional approach
Plate compares, and the conditions such as other marker concentrations, reaction time, cleaning process are consistent.Label is purchased from the U.S. with antibody
Meridian Life Science company, article No. M01246M.Horseradish peroxidase is purchased from Sigma-Aldrich, article No.
P8375.Chemiluminescence detector device is purchased from Beijing Pu Lang Technew SA, instrument model PHX-2012.The purchase of board-washing machine
From Beijing Pu Lang Technew SA, instrument model DNX-9620.Chemiluminescent substrate is purchased from the scorching prosperous biotechnology in Shanghai
Co., Ltd, article No. ECL-P-100.From testing result it can be seen that the detection performance of novel microporous plate be substantially better than tradition it is micro-
Orifice plate.
Detection data under above-mentioned condition is as follows:
Data in table 7 can be seen that novel microporous plate measured value more luminous than the bulk chemical of conventional microplates is considerably higher,
The association reaction of more antigens and antibody has occurred on novel microporous plate.
Data in table 8 can be seen that increasing with concentration of specimens, the increasing degree of the relative deviation of conventional microplates
It is apparently higher than novel microporous plate, the accurate detection range of novel microporous plate is wider than conventional microplates.Concentration of specimens in table is warp
Alpha-fetoprotein antigen concentration in the human serum that commercially available reagent detects.
It is in table 9 statistics indicate that the accuracy of novel microporous plate is substantially better than conventional microporous when detecting low concentration sample
Plate, i.e., novel microporous plate can be such that the detection sensitivity of reagent obviously optimizes.
Claims (11)
1. a kind of for Biological Detection, the solid phase material of analysis, the card including microwell plate or composition microwell plate for detection
Item, it is characterised in that: the detection surface of the microwell plate is fixed with microsphere, and the method that microsphere is fixed on detection surface includes
Following steps: a, by being cleaned and dried to obtain clean detection surface；B, n-isopropyl acrylamide, acrylic acid are dissolved in diformazan
Anthraquinone-2-sodium is added in base sulfoxide, and isopropanol is added, and forms reaction solution, adds microsphere, prepare microsphere reaction solution；
C, above-mentioned microsphere reaction solution is added dropwise on the detection surface of microwell plate, uses ultraviolet light；D, after illumination, successively with
Dehydrated alcohol, distilled water flushing detect surface, to remove unreacted reactant, obtain the microwell plate that microsphere is distributed with after dry.
2. as described in claim 1 a kind of for Biological Detection, the solid phase material of analysis, it is characterised in that: the micropore
The material that plate uses is polystyrene (PS), polytetrafluoroethylene (PTFE) (PTFE), polyvinyl chloride (PVC), polypropylene (PP), polyethylene
(PE), one of silica.
3. a kind of for Biological Detection, the solid phase material of analysis as described in claim 1, it is characterised in that: the microsphere
Material is one of polystyrene (PS), polyethylene (PE), silica.
4. a kind of for Biological Detection, the solid phase material of analysis as claimed in claim 1 or 3, it is characterised in that: the microballoon
The particle size range of body is in 0.02um-5um.
5. a kind of for Biological Detection, the solid phase material of analysis as described in claim 1, it is characterised in that: the microsphere
Surface combined in advance by carboxyl or the chemical group modified of amino, or using antibody, antigen or biological nucleic acid material.
6. a kind of for Biological Detection, the solid phase material of analysis as described in claim 1, it is characterised in that: in the step b
The solution concentration of n-isopropyl acrylamide is 5-10%.
7. a kind of for Biological Detection, the solid phase material of analysis as described in claim 1, it is characterised in that: in the step b
Acrylic acid, solution concentration 1-10%.
8. a kind of for Biological Detection, the solid phase material of analysis as described in claim 1, it is characterised in that: in the step b
Microballoon, solution concentration 1-5mg/ml.
9. a kind of for Biological Detection, the solid phase material of analysis as described in claim 1, it is characterised in that: in the step b
Anthraquinone-2-sodium, solution concentration 0.2-2%.
10. a kind of for Biological Detection, the solid phase material of analysis as described in claim 1, it is characterised in that: the step b
Middle isopropanol, solution concentration 0.1-1.5%.
11. a kind of for Biological Detection, the solid phase material of analysis as described in claim 1, it is characterised in that: the step c
The wavelength of middle ultraviolet light is 300-400nm, and irradiation time is 5-30 minutes.
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|---|---|---|---|---|
| EP1239286A2 (en) * | 2001-03-02 | 2002-09-11 | Hitachi, Ltd. | Biochemical sensors, methods for their manufacture and biochemical methods and systems for testing substances of interest |
| CN2760557Y (en) * | 2004-12-21 | 2006-02-22 | 上海荣盛生物技术有限公司 | Elisa plate with adsorptive force |
| CN1826172A (en) * | 2003-07-23 | 2006-08-30 | 伊斯曼柯达公司 | Random array of microspheres |
| CN1826171A (en) * | 2003-07-23 | 2006-08-30 | 伊斯曼柯达公司 | Colorable microspheres for DNA and protein microarray |
| CN102072955A (en) * | 2010-11-05 | 2011-05-25 | 苏州大学 | Preparation method of modified porous plate |
| CN105556007A (en) * | 2013-09-24 | 2016-05-04 | 恩拓普斯 | Detectable arrays, systems for diagnosis, and methods of making and using the same |
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2017
- 2017-07-28 CN CN201710631319.8A patent/CN107632151B/en not_active Expired - Fee Related
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1239286A2 (en) * | 2001-03-02 | 2002-09-11 | Hitachi, Ltd. | Biochemical sensors, methods for their manufacture and biochemical methods and systems for testing substances of interest |
| CN1826172A (en) * | 2003-07-23 | 2006-08-30 | 伊斯曼柯达公司 | Random array of microspheres |
| CN1826171A (en) * | 2003-07-23 | 2006-08-30 | 伊斯曼柯达公司 | Colorable microspheres for DNA and protein microarray |
| CN2760557Y (en) * | 2004-12-21 | 2006-02-22 | 上海荣盛生物技术有限公司 | Elisa plate with adsorptive force |
| CN102072955A (en) * | 2010-11-05 | 2011-05-25 | 苏州大学 | Preparation method of modified porous plate |
| CN105556007A (en) * | 2013-09-24 | 2016-05-04 | 恩拓普斯 | Detectable arrays, systems for diagnosis, and methods of making and using the same |
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