WO2006051756A1 - カルモデュリンの構造変化を検出する方法、カルモデュリンの構造変化に影響を与える活性を有する物質を探索する方法 - Google Patents
カルモデュリンの構造変化を検出する方法、カルモデュリンの構造変化に影響を与える活性を有する物質を探索する方法 Download PDFInfo
- Publication number
- WO2006051756A1 WO2006051756A1 PCT/JP2005/020377 JP2005020377W WO2006051756A1 WO 2006051756 A1 WO2006051756 A1 WO 2006051756A1 JP 2005020377 W JP2005020377 W JP 2005020377W WO 2006051756 A1 WO2006051756 A1 WO 2006051756A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- calmodulin
- protein
- kinase
- dependent
- sample
- Prior art date
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6872—Intracellular protein regulatory factors and their receptors, e.g. including ion channels
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N19/00—Investigating materials by mechanical methods
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/46—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans from vertebrates
- G01N2333/47—Assays involving proteins of known structure or function as defined in the subgroups
- G01N2333/4701—Details
- G01N2333/4727—Calcium binding proteins, e.g. calmodulin
Definitions
- the present invention relates to a method for detecting a structural change of calmodulin and a method for searching for a substance having an activity that affects the structural change of calmodulin.
- Calmodulin In many intracellular signal transduction systems using calcium as a second messenger, calmodulin first binds to calcium. Calmodulin is a protein with a molecular weight of 16700 consisting of 148 amino acids, and is a calcium-binding protein involved in information transmission functions. One molecule of calmodulin can bind four calciums. It is involved in the regulation of various calmodulin-dependent enzyme Z-calmodulin-binding substances. First, calcium binds to calmodulin with changes in protein conformation, followed by calmodulin-calcium complex. Binds to a calmodulin-dependent enzyme or binding substance and activates the enzyme or binding substance
- the object of the present invention is to detect and measure the structural change of calmodulin in real time using a force sensor as changes in tension and Z or elasticity, so that the structural change of calmodulin can be easily and quickly performed. It is to develop a method to evaluate. Furthermore, by using this method, activity that affects the structural change of calmodulin among many substances can also be obtained. It is also an object of the present invention to enable efficient screening of one ligand.
- the present inventors focused on changes in the tension and / or elasticity of a sample film made of calmodulin when a substance acting as a ligand for calmodulin was added, and as a result of various studies, It was found that the structural change of calmodulin caused by calmodulin ligand can be measured by measuring the tension and / or elasticity of the sample membrane with a force sensor. That is, the present invention forms a sample film made of calmodulin on a substrate, arranges the substrate having the sample film on a force sensor, and applies the test sample to the sample film.
- the present invention provides a method for detecting structural change of calmodulin, which comprises detecting a change in tension and / or elasticity resulting from the change with the force sensor.
- the present invention provides a sample film comprising a calmodulin, a calmodulin fragment, a calmodulin mutant, a tagged calmodulin, or an antibody protein against the calmodulin on a substrate, and the substrate having the sample film is a force sensor.
- the structural change of calmodulin consists of detecting the change in tension and / or elasticity resulting from the structural change of the sample membrane when the test sample is applied to the sample membrane.
- the present invention provides a method for searching for a substance having an activity that affects the above.
- the present invention provides a sample film comprising a calmodulin, a calmodulin fragment, a calmodulin mutant, a tagged calmodulin, or an antibody protein against calmodulin on a substrate, and the substrate having the sample film is a force sensor.
- the calcium / calmodulin consists of detecting the change in tension and Z or elasticity derived from the structural change of the sample membrane when the test sample is applied to the sample membrane.
- the present invention provides a method for searching for a diagnostic or therapeutic agent for a disease in which a change occurs in the intracellular signal transduction pathway involved.
- the present invention adds a substance whose activity is to be determined to a sample film on which calmodulin is immobilized, and can detect a change in tension and / or elasticity of the sample film compared to before adding the substance.
- the present invention provides a method for detecting the structural change of calmodulin. According to the method of the present invention, the activity of the above-mentioned substance causing the structural change of calmodulin can be detected efficiently in a short time in real time.
- FIG. 1 is a graph showing the effects of CaCl and EDTA on calmodulin tension and elastic changes (Example 1).
- FIG. 2 is a graph showing the effect of Ca ion and calmodulin inhibitor W-7 on calmodulin tension and elasticity changes (Example 2).
- FIG. 3 shows the effects of calmodulin binding site fragment of calcium-dependent protein kinase II and Ca ion on calmodulin tension and elasticity changes (Example 3).
- the present invention allows a test substance to act on a calmodulin sample membrane, and detects a change in tension and Z or elasticity resulting from the structural change of the sample membrane by a force sensor.
- This is a method for detecting structural changes in calmodulin. Since the method of the present invention directly observes and measures the structural change of calmodulin in real time as changes in tension and Z or elasticity, the method of the present invention is in that respect that the method of the present invention modifies the activity of force-modulin-dependent enzymes. This is different from the conventional indirect method of measuring the current. Furthermore, using the method of the present invention, a substance having an activity that affects the structural change of calmodulin can also be searched.
- the most prominent feature of the present invention is that the structural change of calmodulin is detected by a change in mechanical properties of a sample film made of calmodulin.
- a sample film made of calmodulin is formed on a substrate.
- the size of the sample film formed is preferably in the range of 50 ⁇ to 1000 zm in the vertical direction, 200 zm to 2000 ⁇ m in the horizontal direction, and 0.3 ⁇ m force to 10 ⁇ m in thickness.
- the substrate in the present invention is an appropriate film support that enables the sample film to be moved to the measuring device by forming the sample film thereon, and the material and size of the substrate.
- the thickness is not particularly limited.
- Calmodulin may be a fragment, mutant or tagged calmodulin as long as it retains the function as calmodulin, which is not necessarily used as a complete protein. That is, in the present specification, “a fragment of calmodulin, a mutant of calmodulin, a tagged calmodulin” means a protein that is a fragment of a part of the amino acid sequence of calmodulin, a protein in which a part of the amino acid sequence of calmodulin is mutated, Alternatively, it is a protein with a tag attached to calmodulin and retains the function of calmodulin by binding to calcium and activating calcium-dependent enzymes.
- the present invention is not limited to forming calmodulin's own sample film, but forms a film made of an antibody protein against calmodulin or a protein activated by calmodulin, and the antibody or calmodulin activating protein Since a similar effect can be obtained by measuring the structural change upon binding to calmodulin, the mode to be applied is also within the scope of the present invention.
- the main calmodulin-dependent enzyme / calmodulin-binding substance located downstream of the calcium calmodulin-mediated signal transduction system and controlled by calmodulin includes adenyl cyclase, brush border myosin I heavy chain, calcineurin, strength Lumodurin-dependent protein kinase II, calmodulin-dependent protein kinase IV, caldesmon, calmodulin-dependent cyclic nucleotide phosphagesterase, erythrocyte calcium-ATPase, neuronal nitric oxide synthase, nicotinamide dinucleotide kinase, faosfati Forces include dilinositol 3 kinase, phosphorylase kinase, skeletal muscle myosin light chain kinase, smooth muscle myosin light chain kinase, IQ GAP1, etc. It is not something.
- calmodulin-dependent enzyme / calmodulin-binding substance
- the ligand that acts on the complex of calmodulin-dependent enzyme It is also possible to That is, a sample membrane made of calmodulin is treated with a calmodulin-dependent enzyme z-calmodulin-binding substance to form a complex, or a sample membrane made of calmodulin-dependent enzyme Z-calmodulin-binding substance is treated with calmodulin to form a complex. It is also an embodiment of the present invention to perform screening on the complex. Considering that the calmodulin-dependent enzyme / calmodulin-binding substance bound to calmodulin controls the downstream signal transduction system, it is preferable for the present invention to form a complex and perform screening. This is an embodiment.
- This calmodulin-dependent enzyme / calmodulin-binding substance does not necessarily need to be a complete protein, so long as it retains the function as a calmodulin-dependent enzyme / binding substance, its fragment, mutant, or tagging It may be a protein. That is, in the present specification, “fragment of calmodulin-dependent enzyme / binding substance, variant of the protein, and tagged protein” means a fragment consisting of a part of the amino acid sequence of the above-mentioned calmodulin-dependent enzyme / binding substance.
- the present invention is not limited to immobilizing the calmodulin-dependent enzyme / calmodulin-binding substance itself, but the same effect can be obtained by immobilizing an antibody against the calmodulin-dependent enzyme / force-modulin-binding substance. Such embodiments are also within the scope of the present invention.
- an electrospray can be used as a method for producing the calmodulin sample film described above.
- the ESD method (electrostatic spray method), in which a sample is deposited by (electrostatic spray) to form a thin film, is suitable.
- electrostatic spray method electrostatic spray method
- Such techniques are well known to those skilled in the art, and can be appropriately modified and used for the purpose of the present invention.
- Japanese Patent Application Laid-Open No. 2003-136005 describes an apparatus for producing a thin film or a spot by fixing it while retaining the activity of a biopolymer or the like.
- Special table 2002-50 Japanese Patent No. 3332 describes a method and apparatus for measuring a ligand that binds to protein or DNA.
- mechanochemically mechanochemically
- an intermediate layer made of a water-soluble polymer between the substrate and the sample film is provided.
- a water-soluble polymer 1.2% polybutylpyrrolidone (PVP) is used as such an intermediate layer.
- PVP polybutylpyrrolidone
- the concentration of force S and PVP in which the concentration of PVP is in the range of 0.1% to 5%, preferably 0.3% to 2% is not particularly limited.
- Other water-soluble polymers can also be used.
- the intermediate layer As examples of materials that can be used as the intermediate layer, as described in JP-T-2002-503332, (1) a layer of a water-soluble polymer such as polyacrylamide or polyethylene glycol , (2) a polymer-strength layer with a disulfide bond reduced by mercaptoethanol, (3) a highly dispersed layer of carbon with low adhesion to deposited biological molecules, and (4 ) It is possible to mention a layer having a low melting point carbon polymer conductive composition.
- a water-soluble polymer such as polyacrylamide or polyethylene glycol
- a polymer-strength layer with a disulfide bond reduced by mercaptoethanol (3) a highly dispersed layer of carbon with low adhesion to deposited biological molecules
- a layer having a low melting point carbon polymer conductive composition It is possible to mention a layer having a low melting point carbon polymer conductive composition.
- calmodulin constituting the sample film may be further crosslinked after immobilization by the ESD method. It is not essential to perform strong crosslinking, but it is effective for the purpose of maintaining the shape and strength of the sample film.
- Cross-linking reagents that can be used to polymerize biological molecules are well known to those skilled in the art, see, for example, Her thigh nson et al., Imm obilized Affinity Ligana fechmques Academic Press, New York, 1991. Can do.
- Daltaraldehyde is most suitable as a reagent used to cross-link calmodulin.
- zero-length cross-linking reagents such as 1_ethyl_3_ (3-dimethylamino) pyrrolopylcarbodiimide (EDC)
- EDC pyrrolopylcarbodiimide
- DMA dimethyl adipine imidate
- SPDP succinimidyl 3 _ (2-pyridyldithio) propionate
- SPDP 4-Azido _ 2 _Nitrophenyl biocytin
- a trivalent cross-linking reagent such as 4-nitrophenyl ester can be mentioned, but is not limited thereto.
- the time for performing the crosslinking reaction is not particularly limited, and optimum conditions can be appropriately selected within the range of about 0 to 3 hours.
- Preparation of applying the test sample to the sample film by placing the sample film thus prepared in, for example, a detection device described in JP-T-2002-503332 and immersing it in an appropriate buffer solution I do.
- the buffer used here is not limited to those which are generally used in this technical field and can use H-type es buffer or Tris buffer.
- the pH of the buffer is not particularly limited, and an appropriate pH can be appropriately selected within the range of about pH 3 to pH 9.
- the buffer solution may have an appropriate salt strength, and it is a preferred embodiment of the present invention to add about 0.1M sodium chloride to the buffer solution as in the following examples. However, it is considered possible to perform the measurement without adding an electrolyte for imparting salt strength, and the mode to be applied is within the scope of the present invention.
- the electrolyte to be added is not limited to sodium chloride.
- the sample membrane is replaced with a buffer solution containing a test sample, which is an object to be tested, and allowed to act on the sample membrane.
- the tension and Z or elastic change of the sample film before and after the test sample is added are measured with a force sensor to evaluate the effect of calmodulin on the structural change. Changes in tension and / or elasticity can be measured with force sensors, preferably with mechanochemical sensors. It is a particularly preferred aspect of the present invention to perform measurement with a mechanochemical sensor using an apparatus described in JP-T-2002-503332.
- calmodulin tension and Z or elasticity changes can be detected quickly and in real time. Therefore, since the structural change of calmodulin in many samples can be evaluated efficiently, the time spent searching for a substance that inhibits the structural change of calmodulin can be greatly reduced, and it is easy to select from a large number of substances. Substances with strong activity can be selected.
- US6033913 having a mechanochemical sensor, and a 10 mM Hepes pH7.4 buffer solution containing 0.1 M NaCl ( Hereinafter, it was immersed in a buffer solution). Buffer was flowed over the membrane present on the detector at a flow rate of 0.1 to 0.2 mL / min to stabilize the tension. Then, at the same flow rate, a CaCl solution dissolved in the above buffer is flowed, and tension and A change in elasticity was detected (Fig. 1).
- This graph shows that CaCl significantly increases the isotropic tension of the calmodulin membrane.
- this action has been shown to reversibly recover because the buffer solution returns to the state prior to the addition of the CaCl solution.
- the decrease in tension due to the addition of EDTA, a chelator indicates that only a small amount of Ca ions in the buffer can be detected.
- the place where the horizontal line indicating the initial state rises shows the point in time when tension is applied to the sample film, and the place where it vibrates shows the measurement of compliance.
- This example shows the structural change of calmodulin caused by Ca 2+ due to the interaction between calmodulin and CaCl, which can be detected in a few minutes in real time using the present invention. is there.
- the structural changes of calmodulin can be referred to the following review article (Vetter SW and Leclerc, E. (2003). Novel aspects of calmodulin target recognition and activation. Eur. J. Biochem. 270, 404- 414
- a calmodulin membrane produced by the same material and method as in Example 1 was immersed in lOmM MOPS buffer ⁇ 7.2 containing 0.1M KC1 and 10 mM EGTA. Calcium buffer solutions with free calcium ion concentrations of 151 ⁇ 352, 352 ⁇ , and 1360 ⁇ were prepared, and either alone or 50 ⁇ of calmodulin inhibitor W-7 was added to detect changes in tension and elasticity (Fig. 2).
- W-7 alone is a force that generates tension in the calmodulin membrane. In the presence of calcium, it increased the tension more and was restored by removing W-7. Since W-7 is known to bind to calmodulin that has undergone structural changes by calcium and to make the structure compact (Osawa, M. et al, (1999). Evidense ror calmodulin mter-domain compaction m FEBS Letters 442, 173-177), this example shows that the strength of calmodulin membrane due to strength modulin inhibitor corresponds to the structural change of calmodulin, and this system is effective in detecting calmodulin inhibitor. It is shown that.
- a calmodulin membrane produced by the same material and method as in Example 1 was prepared using 0.1M KC1, 10m. It was immersed in lOmM HEPES buffer pH 7.2 containing M EGTA. Then, a solution containing the calmodulin binding site fragment of calcium-dependent protein kinase II was prepared, and this was contacted with a force modulin membrane to detect changes in tension and elasticity (Fig. 3).
- a method for detecting a structural change of calmodulin using a mechanochemical sensor has been provided. Furthermore, a method for searching for a substance having an activity that affects the structural change of calmodulin was provided using the above detection method.
- the method of the present invention is considered to contribute to the purpose of obtaining a therapeutic or diagnostic agent for a disease in which an information transmission system via forceful modulin is involved.
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Cell Biology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Medicinal Chemistry (AREA)
- Food Science & Technology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
Description
Claims
Priority Applications (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CA002586415A CA2586415A1 (en) | 2004-11-09 | 2005-11-07 | A method for detecting conformational change of calmodulin, a method for screening a substance having an activity that affects to conformational change of calmodulin |
EP05800408A EP1811284A4 (en) | 2004-11-09 | 2005-11-07 | METHOD FOR DETERMINING A CALMODULINE STRUCTURE MODIFICATION, METHOD FOR OBTAINING MATERIAL THAT CONCERNS THE CALMODULINE STRUCTURE CHANGE OF ACTIVE ACTIVITY |
US11/666,077 US20080115593A1 (en) | 2004-11-09 | 2005-11-07 | Method for Detecting Conformational Change of Calmodulin, a Method for Screening a Substance Having an Activity that Affects to Conformational Change of Calmodulin |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2004325005A JP2006133164A (ja) | 2004-11-09 | 2004-11-09 | カルモデュリンの構造変化を検出する方法、カルモデュリンの構造変化に影響を与える活性を有する物質を探索する方法 |
JP2004-325005 | 2004-11-09 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2006051756A1 true WO2006051756A1 (ja) | 2006-05-18 |
Family
ID=36336437
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/JP2005/020377 WO2006051756A1 (ja) | 2004-11-09 | 2005-11-07 | カルモデュリンの構造変化を検出する方法、カルモデュリンの構造変化に影響を与える活性を有する物質を探索する方法 |
Country Status (5)
Country | Link |
---|---|
US (1) | US20080115593A1 (ja) |
EP (1) | EP1811284A4 (ja) |
JP (1) | JP2006133164A (ja) |
CA (1) | CA2586415A1 (ja) |
WO (1) | WO2006051756A1 (ja) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR101187674B1 (ko) | 2010-09-10 | 2012-10-10 | 송인국 | 이단 면역 크로마토그래피를 이용한 효소 활성 측정법 |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH05508079A (ja) * | 1991-04-19 | 1993-11-18 | ザ ボード オブ リージェンツ オブ ザ ユニヴァーシティ オブ ワシントン | 哺乳類ホスホジエステラーゼをコードするdna |
JP2002503332A (ja) * | 1996-06-20 | 2002-01-29 | ニューヨーク ユニヴァーシティ | ポリマ材料と相互作用するリガンドの検出 |
JP2003522946A (ja) * | 2000-02-10 | 2003-07-29 | エボテック オーアーイー アクチェンゲゼルシャフト | 蛍光強度及び寿命分布分析 |
JP2004510145A (ja) * | 2000-09-20 | 2004-04-02 | モレキュラー・リフレクションズ | 共鳴センサーとして使用するための微細加工された超音波アレイ |
Family Cites Families (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0988112B1 (en) * | 1997-06-20 | 2010-04-14 | New York University | Electrospraying solutions of substances for mass fabrication of chips and libraries |
AU2001280552A1 (en) * | 2000-07-13 | 2002-01-30 | The Ohio State University Research Foundation | Multimeric biopolymers as structural elements, sensors and actuators in microsystems |
KR100442822B1 (ko) * | 2001-10-23 | 2004-08-02 | 삼성전자주식회사 | 전단응력 측정을 이용한 생분자들간의 결합 여부 검출 방법 |
CA2463301A1 (en) * | 2004-04-22 | 2005-10-22 | Atanas Iliev Lalev | Method for detecting protein interactions - affinity purification with switching the ligand |
CA2561246A1 (en) * | 2004-03-25 | 2005-10-06 | Fuence Co., Ltd. | Method of detecting conformational change of an amyloid protein, a method of searching a substance having an activity that affects to conformational change of an amyloid protein, and a method of searching a therapeutic or diagnostic agent for amyloid-related diseases |
JP2006090782A (ja) * | 2004-09-22 | 2006-04-06 | Fyuuensu:Kk | 基板上に固定化した蛋白質の構造変化を検出する方法 |
-
2004
- 2004-11-09 JP JP2004325005A patent/JP2006133164A/ja active Pending
-
2005
- 2005-11-07 WO PCT/JP2005/020377 patent/WO2006051756A1/ja active Application Filing
- 2005-11-07 US US11/666,077 patent/US20080115593A1/en not_active Abandoned
- 2005-11-07 CA CA002586415A patent/CA2586415A1/en not_active Abandoned
- 2005-11-07 EP EP05800408A patent/EP1811284A4/en not_active Withdrawn
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH05508079A (ja) * | 1991-04-19 | 1993-11-18 | ザ ボード オブ リージェンツ オブ ザ ユニヴァーシティ オブ ワシントン | 哺乳類ホスホジエステラーゼをコードするdna |
JP2002503332A (ja) * | 1996-06-20 | 2002-01-29 | ニューヨーク ユニヴァーシティ | ポリマ材料と相互作用するリガンドの検出 |
JP2003522946A (ja) * | 2000-02-10 | 2003-07-29 | エボテック オーアーイー アクチェンゲゼルシャフト | 蛍光強度及び寿命分布分析 |
JP2004510145A (ja) * | 2000-09-20 | 2004-04-02 | モレキュラー・リフレクションズ | 共鳴センサーとして使用するための微細加工された超音波アレイ |
Non-Patent Citations (1)
Title |
---|
See also references of EP1811284A4 * |
Also Published As
Publication number | Publication date |
---|---|
US20080115593A1 (en) | 2008-05-22 |
EP1811284A4 (en) | 2008-12-24 |
CA2586415A1 (en) | 2006-05-18 |
JP2006133164A (ja) | 2006-05-25 |
EP1811284A1 (en) | 2007-07-25 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Dechtrirat et al. | Protein rebinding to a surface‐confined imprint | |
US20230084931A1 (en) | Methods and systems for characterizing analytes using nanopores | |
Fischer | Amine coupling through EDC/NHS: a practical approach | |
Haji-Hashemi et al. | Simple and effective label free electrochemical immunosensor for Fig mosaic virus detection | |
US20140342359A1 (en) | Versatile and sensitive biosensor | |
JP2003508763A (ja) | 固体支持体上にリガンドを固定化する方法及び装置並びにその使用方法 | |
WO2014130793A1 (en) | Epitope-polymer platform for detection of bacterial organisms | |
Liu et al. | Selective detection of 8-Oxo-2′-deoxyguanosine in single-stranded DNA via nanopore sensing approach | |
EP4211259A1 (en) | Electrochemical biosensor for target analyte detection | |
Ballestar et al. | Conformational changes in the nucleosome followed by the selective accessibility of histone glutamines in the transglutaminase reaction: effects of ionic strength | |
JPS59224564A (ja) | 生物学的活性もしくは官能性化合物の複合化およびその調製およびその利用 | |
WO2006051756A1 (ja) | カルモデュリンの構造変化を検出する方法、カルモデュリンの構造変化に影響を与える活性を有する物質を探索する方法 | |
JP4940456B2 (ja) | 標的に対して高い親和性で結合する核酸を選択する方法 | |
JP2005500053A5 (ja) | ||
WO2006136686A1 (fr) | Procede d'immobilisation de l'adn superenroule et utilisation pour analyser la reparation de l'adn | |
WO2012031294A2 (en) | Methods for determining protein ligand binding | |
Singh et al. | Small molecule microarray screening methodology based on surface plasmon resonance imaging | |
Ivan et al. | Competitive ligands facilitate dissociation of the complex of bifunctional inhibitor and protein kinase | |
US20090068673A1 (en) | Utilization of compatible solutes to improve the performance of the techniques using immobilized biologic materials | |
Hao et al. | Highly sensitive poly-N-isopropylacrylamide microgel-based electrochemical biosensor for the detection of SARS-COV-2 spike protein | |
Ibanez et al. | DNA detection using a triple readout optical/AFM/MALDI planar microwell plastic chip | |
Al Borhani et al. | Selection of a new aptamer targeting amoxicillin for utilization in a label-free electrochemical biosensor | |
JP2003502665A (ja) | 検出可能な競合リガンドを使用した、親和性リガンドのスクリーニングに適したキャピラリ電気泳動法 | |
Rich et al. | Extracting affinity constants from biosensor binding responses | |
Yoo et al. | A radioimmunoassay method for detection of DNA based on chemical immobilization of anti-DNA antibody |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AK | Designated states |
Kind code of ref document: A1 Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BW BY BZ CA CH CN CO CR CU CZ DE DK DM DZ EC EE EG ES FI GB GD GE GH GM HR HU ID IL IN IS KE KG KM KN KP KR KZ LC LK LR LS LT LU LV LY MA MD MG MK MN MW MX MZ NA NG NI NO NZ OM PG PH PL PT RO RU SC SD SE SG SK SL SM SY TJ TM TN TR TT TZ UA UG US UZ VC VN YU ZA ZM ZW |
|
AL | Designated countries for regional patents |
Kind code of ref document: A1 Designated state(s): GM KE LS MW MZ NA SD SL SZ TZ UG ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IS IT LT LU LV MC NL PL PT RO SE SI SK TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG |
|
121 | Ep: the epo has been informed by wipo that ep was designated in this application | ||
WWE | Wipo information: entry into national phase |
Ref document number: 11666077 Country of ref document: US |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2586415 Country of ref document: CA |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2005800408 Country of ref document: EP |
|
WWP | Wipo information: published in national office |
Ref document number: 2005800408 Country of ref document: EP |
|
WWP | Wipo information: published in national office |
Ref document number: 11666077 Country of ref document: US |