IE20170054A1 - Method of testing surface plasmon resonance sensor - Google Patents
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- IE20170054A1 IE20170054A1 IE20170054A IE20170054A IE20170054A1 IE 20170054 A1 IE20170054 A1 IE 20170054A1 IE 20170054 A IE20170054 A IE 20170054A IE 20170054 A IE20170054 A IE 20170054A IE 20170054 A1 IE20170054 A1 IE 20170054A1
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- 238000002198 surface plasmon resonance spectroscopy Methods 0.000 title claims abstract description 41
- 238000010998 test method Methods 0.000 title claims abstract description 16
- 239000003446 ligand Substances 0.000 claims abstract description 54
- 230000027455 binding Effects 0.000 claims abstract description 44
- 230000009871 nonspecific binding Effects 0.000 claims abstract description 26
- 239000000463 material Substances 0.000 claims abstract description 21
- 239000002082 metal nanoparticle Substances 0.000 claims abstract description 21
- 238000000034 method Methods 0.000 claims abstract description 16
- 230000008859 change Effects 0.000 claims abstract description 14
- 239000007853 buffer solution Substances 0.000 claims description 13
- 238000005259 measurement Methods 0.000 abstract description 18
- 238000004519 manufacturing process Methods 0.000 abstract description 16
- 108090000623 proteins and genes Proteins 0.000 abstract description 14
- 102000004169 proteins and genes Human genes 0.000 abstract description 14
- 230000009870 specific binding Effects 0.000 abstract description 11
- 238000011065 in-situ storage Methods 0.000 abstract description 5
- 239000000243 solution Substances 0.000 description 26
- 239000002105 nanoparticle Substances 0.000 description 11
- 239000012491 analyte Substances 0.000 description 10
- 238000001179 sorption measurement Methods 0.000 description 10
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 9
- 229910052737 gold Inorganic materials 0.000 description 9
- 239000010931 gold Substances 0.000 description 9
- 239000000427 antigen Substances 0.000 description 8
- 238000010586 diagram Methods 0.000 description 7
- 102000036639 antigens Human genes 0.000 description 5
- 108091007433 antigens Proteins 0.000 description 5
- 238000006243 chemical reaction Methods 0.000 description 5
- 230000003287 optical effect Effects 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- 229940098773 bovine serum albumin Drugs 0.000 description 4
- 238000003780 insertion Methods 0.000 description 4
- 230000037431 insertion Effects 0.000 description 4
- 238000001514 detection method Methods 0.000 description 3
- 239000003550 marker Substances 0.000 description 3
- 239000013307 optical fiber Substances 0.000 description 3
- 238000012545 processing Methods 0.000 description 3
- 206010060862 Prostate cancer Diseases 0.000 description 2
- 208000000236 Prostatic Neoplasms Diseases 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 239000002184 metal Substances 0.000 description 2
- 229910052751 metal Inorganic materials 0.000 description 2
- 238000003908 quality control method Methods 0.000 description 2
- 239000002094 self assembled monolayer Substances 0.000 description 2
- 239000013545 self-assembled monolayer Substances 0.000 description 2
- 229910052709 silver Inorganic materials 0.000 description 2
- 239000004332 silver Substances 0.000 description 2
- 239000000439 tumor marker Substances 0.000 description 2
- 238000012935 Averaging Methods 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 239000003574 free electron Substances 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000010409 thin film Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
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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
- G01N21/554—Attenuated total reflection and using surface plasmons detecting the surface plasmon resonance of nanostructured metals, e.g. localised surface plasmon resonance
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/403—Cells and electrode assemblies
- G01N27/404—Cells with anode, cathode and cell electrolyte on the same side of a permeable membrane which separates them from the sample fluid, e.g. Clark-type oxygen sensors
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- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Pathology (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Electrochemistry (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Nanotechnology (AREA)
- Investigating Or Analysing Materials By Optical Means (AREA)
Abstract
Provided is a method of testing a localized surface plasmon resonance sensor based on a surface plasmon resonance phenomenon of metal nanoparticles. The method includes binding a material for preventing nonspecific binding to metal nanoparticles with a ligand thereon, and deriving a change (D) in sensor output signal intensity caused by the binding of the material for preventing nonspecific binding; and evaluating conditions of ligand binding based on D. The method can be carried out in-situ during the manufacturing of a biosensor or the measurement for specific binding of proteins or measurement, and thus is capable of determining sensor performance or adequacy in a convenient manner.
Description
l7l3$5é OF TESTING SURFACE PLASMON RESONANCE SENSOR CLAIM FOR PRIORITY This application claims priority to Korean Patent Application No. 2016-0168829 filed on Dec. 12, 2016 in the Korean Intellectual Property Office (KIPO), the entire contents of which are hereby incorporated by reference.
BACKGROUND 1. Technical Field 10 Example embodiments of the present invention relate in general to the field of surface plasmon resonance sensors and more specifically to a method of testing a surface plasmon resonance sensor, the method capable of in-situ testing during a sensor manufacturing process. 2. Related Art 15 Surface plasmon resonance is a phenomenon caused by collective vibration of free electrons when incident light reacts with nanoparticles or a metal thin film of gold or silver.
Having an advantage of ability to measure reactions among biomaterials in real time without requiring a specific marker, surface plasmon resonance has been applied for biosensors capable of measuring various bioreactions and protein chip analyses. 20 When perfomiing biological measurement such as the measuring of specific binding among proteins using a surface plasmon resonance sensor, a ligand is adsorbed onto a surface of the sensor, and a sensor output signal is detected as analytes react with the ligand. In this case, the reaction between the analytes and the ligand adsorbed onto the sensor surface is significantly affected by the amount, chemical stability, and the like of the ligand. 25 Therefore, to analyze analyte detection characteristics such as sensitivity, reproducibility, and stability and evaluate overall sensor performance based on the analyte detection characteristics, proper observation, response, and analysis of conditions of ligand adsorption onto the sensor surface need to be carried out from the sensor manufacturing stage.
Most sensors currently in use lack a method enabling the understanding of ligand adsorption during a sensor manufacturing stage and a sensor preparation stage. Generally, ligand adsorption is confirmed through analysis according to a method (e.g. sandwich assay) involving the double binding of the analyte with the above ligand and another ligand linked with a proper label or marker such that the analyte is sandwiched between the above two types of ligands. However, such a method may be considered as the manufacturing of a new type of a sensor for analyzing ligand adsorption, and requires the use of additional analytical equipment capable of measuring the label or marker after all processing is completed.
Also from the perspective of a sensor manufacturer, conditions of ligand adsorption need to be analyzed in-situ during the sensor manufacturing stage for sensor performance and quality control, but there is no known proper method for realizing this need.
I Conventional Art Documents) [Patent document] (Patent document 1) Korean Patent Application No. 10-2002-0067661 SUMMARY Accordingly, example embodiments of the present invention are provided to substantially obviate one or more problems due to limitations and disadvantages of the related art, and provide a method of testing a surface plasmon resonance sensor in-situ during a sensor device manufacturing process.
Example embodiments of the present invention provide a test method capable of evaluating conditions of ligand binding during a measuring or manufacturing process of a localized surface plasmon resonance sensor.
In some example embodiments, a method of testing a localized surface plasmon resonance sensor based on a surface plasmon resonance phenomenon of metal nanoparticles is provided. The method includes: binding a material for preventing nonspecific binding to the metal nanoparticles in the localized surface plasmon resonance sensor wherein the metal nanoparticles have a ligand bound thereon; deriving a change D in sensor output signal intensity caused by the binding of the material for preventing nonspecific binding; and evaluating conditions of binding of the ligand based on D.
The conditions of binding of the ligand may be evaluated by setting a reference range for a change in sensor output signal intensity caused by the binding of the material for preventing nonspecific binding in advance and then comparing the change D to the reference range.
The change D in sensor output signal intensity caused by the binding of the material for preventing nonspecific binding is a difference in sensor output signals that are obtained based on light reflected from the metal nanoparticles with the ligand bound thereon in a buffer solution respectively before and after the binding of the material for preventing nonspecific binding.
The change D being greater than the reference range is considered to indicate inappropriate ligand binding due to a shortage or instability of the ligand engaged in binding.
Example embodiments of the present invention provide an in-situ test method capable of analyzing conditions of a ligand bound to metal nanoparticles of a localized surface plasmon resonance sensor during sensor manufacturing or measurement. The performance or adequacy of the sensor can be determined in a simple manner by analyzing conditions of ligand binding during the course of preparing a sensor device or system capable of measuring actual specific binding among proteins. In this way, a more reliable measurement result can be obtained from the sensor.
BRIEF DESCRIPTION OF DRAWINGS Example embodiments of the present invention will become more apparent by describing in detail example embodiments of the present invention with reference to the accompanying drawings, in which: FIG. 1 is a diagram and images for illustrating a process of manufacturing a fiber- optic localized surface plasmon resonance sensor to which a test method of a surface plasmon resonance sensor according to an example embodiment of the present invention is applied; FIG. 2 is a diagram for illustrating an example surface plasmon resonance sensor device to which a test method according to an example embodiment of the present invention is applied; FIG. 3 is a diagram for illustrating a fiber-optic surface plasmon resonance sensor included in the sensor device of FIG. 2; and FIGS. 4 and 5 are diagrams provided for describing a process of testing a surface plasmon resonance sensor according to an example embodiment of the present invention.
DESCRIPTION OF EXAMPLE EMBODIMENTS Hereinafter, example embodiments of the present invention will be described in detail with reference to the accompanying drawings. In describing example embodiments of the present invention, detailed descriptions of related well-known functions or configurations deemed to unnecessarily obscure the gist of the present invention will be omitted.
Example embodiments of the present invention provide a test method capable of analyzing conditions of ligand binding in a localized surface plasmon resonance sensor.
Such a test may be carried out during a manufacturing process of a sensor device including the sensor. The performance or adequacy of a surface plasmon resonance sensor may be determined in a simple manner, for example, by analyzing conditions of ligand binding during the course of preparing a sensor device or system that includes the surface plasmon resonance sensor containing gold nanoparticles adsorbed onto the core of optical fiber ends and is capable of measuring actual specific binding among proteins. In this way, a more reliable measurement result may be obtained from the sensor, fed back to the sensor manufacturing process, and thus be reflected in sensor quality control.
Biological measurement such as the measuring of specific binding among proteins using a localized surface plasmon resonance sensor involves adsorbing a ligand onto metal nanoparticles and detecting a sensor output signal while inducing an analyte reaction with the ligand. To carry out such measurement, a hardware arrangement is implemented, which may include a sensor, a solution supply unit for providing solutions required for the measurement, a component for causing the supplied solutions to contact metal nanoparticles, an optical measuring unit, and the like. Then, a ligand required for the measurement is bound to metal nanoparticles by allowing a solution containing the ligand to contact the nanoparticles, and subsequently, a solution containing analytes is allowed to contact the ligand. Then, specific binding is analyzed.
FIG. 1 is a diagram and images for illustrating a process of manufacturing a fiber- optic localized surface plasmon resonance sensor to which a test method of a surface plasmon resonance sensor according to an example embodiment of the present invention is applied.
FIG. 2 is a diagram for illustrating an example surface plasmon resonance sensor device to which a test method according to an example embodiment of the present invention is applied.
FIG. 3 is a diagram for illustrating a fiber-optic surface plasmon resonance sensor included in the sensor device of FIG. 2.
Referring to the drawings, the method of testing a surface plasmon resonance sensor according to example embodiments of the present invention may be suitably applied to the fiber-optic localized surface plasmon resonance sensor as illustrated in FIGS. 1 to 3, but is not limited thereto.
The fiber-optic localized surface plasmon resonance sensor may be realized, for example, by using a self-assembled monolayer (SAM) to bind metal (e.g. gold or silver) nanoparticles to the core region of optical fiber ends in a manner as shown in FIG. 1.
Sensor device hardware arrangement is prepared for the measuring of the above surface plasmon resonance sensor in a manner as shown in FIG. 2. For example, the sensor hardware arrangement may include a microchannel unit 2 including a microchannel having a plurality of solution inlets 21 and a solution outlet 22; a solution supply unit 3 for supplying individual solutions into the solution inlets 21; a surface plasmon resonance sensor 1 installed within the microchannel unit 2; and an optical measuring unit 4 connected to a rear end of the sensor. Such a configuration of the sensor device makes it possible to minimize the mixing of the plurality of solutions and allow the solutions to flow in a sequential manner for the measurement.
To measure specific binding of a specific analyte, the analyte is released while a ligand (antibody) is bound to metal nanoparticles of a sensor installed within the aforementioned sensor device, and an antibody-antigen reaction is measured. The process of measuring specific binding among proteins or the like is generally indicated by way of a sensorgram, which indicates the magnitude of output signals of a sensor over time in a continuous manner.
As shown in FIGS. 4 and 5 which illustrate the measurement of a prostate cancer marker as exemplary measurement, the test method according to example embodiments of the present invention is carried out by measuring and recording sensor output signals over time while immersing the sensor in a buffer solution, a ligand solution, a buffer solution, a solution containing a protein for preventing nonspecific binding, a buffer solution, and an antigen solution in the written order.
According to the test method of example embodiments of the present invention, sensor output signal intensity is measured both before and after binding the material for preventing nonspecific binding to the ligand—bound metal nanoparticles while an end portion of the sensor (i.e. the portion that contains metal nanoparticles) is immersed in a buffer solution, and a change D in output signal intensity due to the binding of the material for preventing nonspecific binding is derived from a difference in the sensor output signal intensities measured as above. D may be used to determine conditions of ligand binding.
In this case, the material or protein for preventing nonspecific binding inhibits nonspecific binding by adsorbing onto a ligand-free region of a surface of the metal nanoparticles in the sensor. Therefore, with a larger portion of the ligand being adsorbed onto the surface of the metal nanoparticles in a stable manner, less material for preventing nonspecific binding is engaged in binding. Conversely, when a smaller portion of the ligand is adsorbed onto the surface of the metal nanoparticles or the adsorption is unstable enough to allow easy separation, more protein for preventing nonspecific binding will be engaged in binding. In other words, a larger D may be considered to indicate the binding of a smaller amount of the ligand to the surface of the metal nanoparticles in the sensor as well as unstable binding, whereas a smaller D may be considered to indicate the binding of a larger amount of the ligand as well as stable binding.
For example, such a consideration of conditions of ligand binding may be made by setting a reference range or value in advance, and then comparing measured values with the reference range or value. The reference range or value may be derived by averaging accumulated measurement results.
The change D in sensor output signal intensity caused by the binding of the material for preventing nonspecific binding may be derived from a sensorgram in a simple manner without requiring specific processing or measuring devices, wherein the sensorgram has been obtained based on the specific binding of a detection target during the manufacturing process of the sensor device. Sensor output signal intensity B is measured in a buffer solution following ligand binding, and sensor output signal intensity C is determined in a buffer solution following the binding of the material for preventing nonspecific binding. A difference between B and C (i.e. C-B) is calculated.
Example The localized surface plasmon resonance sensor as shown in FIG. 1 is realized by causing gold nanoparticles to be adsorbed onto an end of an optical fiber.
Sensor device hardware arrangement including a fiber-optic surface plasmon resonance sensor 1, a microchannel unit 2, a solution supply unit 3, and an optical measuring unit 4 is implemented.
For example, the microchannel unit 2 includes a microfluidic channel and a sensor insertion hole 23, wherein the microfluidic channel includes a plurality of solution inlets 21 and a solution outlet 22, and the sensor insertion hole 23 is a hole into which a fiber-optic surface plasmon resonance sensor 1 is inserted. The surface plasmon resonance sensor 1 is inserted into the sensor insertion hole 23 in such a way that a front end portion thereof is exposed within the microfluidic channel such that gold nanoparticles immobilized to a core surface Contact a fluid in the channel. A rear end portion of the surface plasmon resonance sensor 1 is connected with a light source 41 and a detector 42 of the optical measuring unit 4 via a coupler 43.
An antibody-antigen reaction of a prostate cancer marker is measured. In this case, the antibody corresponds to a ligand and the antigen corresponds to an analyte. As shown in FIG. 3, the sensor for measuring the antigen as an analyte is configured to include the antibody (i.e. ligand) adsorbed onto a surface of gold nanoparticles and a protein for preventing nonspecific binding adsorbed onto the nanoparticles. As the material for preventing nonspecific binding, bovine serum albumin (BSA) is used.
Specifically, first, a sensorgram based on measured sensor output signals is plotted while supplying a DI water or a buffer solution, a ligand solution, a buffer solution, a solution containing the material for preventing nonspecific binding, and a buffer solution in the written order through respectively designated solution inlets 21 from the solution supply unit 3. To derive a final measurement value for detecting final specific binding of proteins and the like, a test according to example embodiments of the present invention is carried out prior to supplying a solution containing the antigen.
Conditions of a surface of gold nanoparticles during the course of sensor manufacturing and analyte measurement may be illustrated as shown in FIG. 4 or indicated as a sensorgram as provided in FIG. 5.
As shown in FIGS. 4 and 5, conditions of binding of the ligand according to example embodiments of the present invention are understood or determined by causing BSA, which is a material for preventing nonspecific binding, to bind to ligand-bound gold nanoparticles and then deriving a change D in sensor output signal intensity due to such binding. As shown in FIG. 5, D is a difference between B and C (i.e. C-B), which are sensor output signals following the supply of a buffer solution into the microchannel unit 2 and are measured respectively before and after BSA binding.
Conditions of ligand adsorption may be analyzed based on the degree of adsorption of a protein for preventing nonspecific binding and derived using D. In other words, D being significantly increased compared to other values, the predetermined reference value, or the predetermined reference range indicates inappropriate adsorption of the antibody (i.e. ligand) onto a sensor surface. In contrast, D being smaller than other values, the reference range, or the reference value indicates stable adsorption of the ligand. Such analysis is performed based on a sensorgram that indicates a series of processes for sensor manufacturing and measurement, and thus can be implemented in a simple manner without requiring additional processing or devices.
For reference, a final measurement value for detecting specific binding of proteins is represented as F in FIG. 5 and corresponds to a change F in sensor output signal intensity caused by antibody-antigen binding. F is a difference between C and E (i.e. E-C), which are obtained based on light reflected from gold nanoparticles of the sensor following the supply of a buffer solution and are measured respectively before and after binding the antigen to the antibody.
While the example embodiments of the present invention and their advantages have been described in detail, it should be understood that various changes, substitutions and alterations may be made herein without departing from the scope of the invention. 5 [Description of Reference Numerals] 1: sensor 2: microchannel unit 3: solution supply unit 4: optical measuring unit 21: solution inlets 22: solution outlet 10 23: sensor insertion hole 41: light source 42: detector 43: coupler
Claims (4)
1. A method of testing a localized surface plasmon resonance sensor based on a surface plasmon resonance phenomenon of metal nanoparticles, comprising: binding a material for preventing nonspecific binding to the metal nanoparticles in the localized surface plasmon resonance sensor wherein the metal nanoparticles have a ligand bound thereto; deriving a change D in sensor output signal intensity caused by the binding of the material for preventing nonspecific binding; and evaluating conditions of binding of the ligand based on D.
2. The method of claim 1, wherein the conditions of binding of the ligand may be evaluated by setting a reference range for a change in sensor output signal intensity caused by the binding of the material for preventing nonspecific binding in advance and then comparing the change D to the reference range.
3. The method of claim 1, wherein the change D in sensor output signal intensity caused by the binding of the material for preventing nonspecific binding is a difference in sensor output signals that are obtained based on light reflected from the metal nanoparticles with the ligand bound thereon in a buffer solution respectively before and after the binding of the material for preventing nonspecific binding.
4. The method of claim 2, wherein the D being greater than the reference range is considered to indicate inappropriate ligand binding due to a shortage or instability of the ligand engaged in binding. -2 l70€l;‘:a a
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020160168829A KR101881228B1 (en) | 2016-12-12 | 2016-12-12 | Method for testing surface plasmon resonance sensor |
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| IE20170054A1 true IE20170054A1 (en) | 2018-08-08 |
| IE87052B1 IE87052B1 (en) | 2019-10-02 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| KR102233028B1 (en) * | 2019-08-06 | 2021-03-29 | 단국대학교 산학협력단 | Surface plasmon resonance sensor combined with nanostructure |
| KR102233036B1 (en) * | 2019-08-06 | 2021-03-29 | 단국대학교 산학협력단 | Continuous measurement device using surface plasmon resonance sensor |
| KR102233031B1 (en) * | 2019-08-06 | 2021-03-29 | 단국대학교 산학협력단 | Test method for surface plasmon resonance sensor combined with nanostructures, and calibration method of output signal of surface plasmon resonance sensor combined with nanostructure |
| KR20250010847A (en) | 2023-07-13 | 2025-01-21 | 단국대학교 산학협력단 | Multibranched metal nanoparticle and biosensor based upon sandwich assay using it |
| KR20250011594A (en) | 2023-07-14 | 2025-01-21 | 단국대학교 산학협력단 | Method for manufacturing fiber optic based plasmonic biosensor for dementia diagnosis and biosensor manufactured therefrom |
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| US6378566B1 (en) | 2001-02-16 | 2002-04-30 | Albany International Corp. | Enhancements for seams in on-machine-seamable papermaker's fabrics |
| FI118061B (en) | 2001-09-24 | 2007-06-15 | Beanor Oy | Procedure and bio donor for analysis |
| JP2006322878A (en) | 2005-05-20 | 2006-11-30 | Matsushita Electric Ind Co Ltd | Method for analyzing ligand in sample and apparatus for analyzing ligand in sample |
| KR20100067016A (en) * | 2008-12-10 | 2010-06-18 | 한국전자통신연구원 | Apparatus for detecting bio materials and method for detecting bio materials by using the apparatus |
| JP2015064205A (en) | 2013-09-24 | 2015-04-09 | スタンレー電気株式会社 | Localized plasmon resonance chip and localized plasmon resonance biosensor using the same |
| KR20150138968A (en) * | 2014-05-30 | 2015-12-11 | 재단법인대구경북과학기술원 | Plate for diagnosis and manufacturing method thereof |
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| IE87052B1 (en) | 2019-10-02 |
| KR101881228B1 (en) | 2018-08-17 |
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