WO2001063283A1 - Procede de mesure d'une substance echantillon physiologiquement active au moyen d'un filtre poreux - Google Patents

Procede de mesure d'une substance echantillon physiologiquement active au moyen d'un filtre poreux Download PDF

Info

Publication number
WO2001063283A1
WO2001063283A1 PCT/JP2001/001196 JP0101196W WO0163283A1 WO 2001063283 A1 WO2001063283 A1 WO 2001063283A1 JP 0101196 W JP0101196 W JP 0101196W WO 0163283 A1 WO0163283 A1 WO 0163283A1
Authority
WO
WIPO (PCT)
Prior art keywords
substance
ligand
solution
sample
physiologically active
Prior art date
Application number
PCT/JP2001/001196
Other languages
English (en)
Japanese (ja)
Inventor
Kiyosato Niu
Yoshinori Suzuki
Toshio Tajima
Tsuneo Hanyu
Masaru Tanebe
Original Assignee
Nippon Chemiphar Co., Ltd.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Nippon Chemiphar Co., Ltd. filed Critical Nippon Chemiphar Co., Ltd.
Priority to KR1020027010881A priority Critical patent/KR20020086558A/ko
Priority to AU2001232350A priority patent/AU2001232350A1/en
Publication of WO2001063283A1 publication Critical patent/WO2001063283A1/fr

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
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/536—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase
    • G01N33/537—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase with separation of immune complex from unbound antigen or antibody
    • G01N33/538—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase with separation of immune complex from unbound antigen or antibody by sorbent column, particles or resin strip, i.e. sorbent materials
    • 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
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • 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
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54306—Solid-phase reaction mechanisms

Definitions

  • the present invention relates to a method for measuring the amount of a sample substance having a physiological activity based on immunoassay using a porous filter as a solid support.
  • an immobilized antibody solid phase corresponding to the relevant antigen or an immobilized antigen solid phase corresponding to the relevant antibody is prepared in advance according to the antigen or antibody to be measured, and the measurement is performed.
  • the method of selecting the solid phase corresponding to the target substance has been generally used each time. In such a method, the number of types of solid phase required for each substance to be measured is innumerable, so that the preparation of the solid phase is extremely complicated, and the measurement operation is performed mechanically or irrespective of the method used. The complexity of having to select an appropriate solid phase depending on the target substance was added.
  • the problem to be solved by the present invention is to use the conventional immunoassay A method for measuring biologically active sample substances based on immunoassays that can obtain measurement sensitivity at the level of nanograms per milliliter in a time that is significantly shorter than the time required for immunoassays. To provide.
  • the present invention provides a sample solution containing a measurement sample substance having a physiological activity, and a solution containing a conjugate in which a ligand is bound to a first physiologically active substance that specifically binds to the measurement sample substance.
  • the solution is brought into contact with an amount such that the volume ratio of the solution is 1: 1 to 20: 1 (preferably, 2: 1 or more, and 10: 1 or less).
  • Forming a complex of the substance and the measurement sample substance in a solution dropping a solution containing the complex onto the surface of the porous filter to which the ligand capturing agent is bound, A solution containing an enzyme-labeled second physiologically active substance, which specifically binds to the measurement sample substance, on the surface of the porous filter.
  • the method for measuring the amount of a biologically active sample substance using the porous filter As the porous filter used in the measurement method of the present invention, a fibrous porous filter is preferable, but a porous ceramic filter can also be used.
  • the present invention also provides a sample solution containing a measurement sample substance having a physiological activity, a solution containing a conjugate in which a ligand is bound to a first physiologically active substance that specifically binds to the measurement sample substance, and the measurement sample
  • a solution containing an enzyme-labeled second physiologically active substance that specifically binds to a substance is mixed with the sample solution and the conjugate solution at a volume ratio of 1: 1 to 20: 1 (preferably, (2: 1 or more, and 10: 1 or less), the first physiologically active substance bound to the ligand, the sample substance to be measured, and the enzyme-labeled physiologically active substance.
  • a complex in a solution Forming a complex in a solution; dropping the solution containing the complex onto the surface of the porous filter to which the ligand scavenger is bound; Binding the ligand portion of the complex to the ligand scavenger; removing unbound enzyme-labeled second physiologically active substance by washing the porous filter;
  • a method for measuring the amount of a physiologically active sample substance using a porous filter which comprises a step of measuring the activity of an enzyme bound to the enzyme.
  • a fibrous porous filter is preferable, but a porous ceramic filter can also be used.
  • the ligand capture agent is an anti-biotin antibody, and the ligand is an antibody against the anti-biotin antibody.
  • the ligand capture agent is streptavidin.
  • the ligand capture agent is streptavidin, and the ligand is an anti-streptavidin antibody.
  • the ligand-capturing agent is streptavidin
  • the spacer is an antibody against an anti-streptavidin antibody and an anti-streptavidin antibody layered thereon.
  • Absorber is provided at the bottom of the fibrous porous filter.
  • a fibrous porous filter is used as a solid support, and a ligand-capturing agent for capturing a ligand is supported on the solid support, and is used as a first physiologically active substance that specifically binds to a measurement sample substance.
  • a certain amount of the reaction liquid is directly dropped onto the solid phase carrier to which the ligand capture agent is bound.
  • a solution containing an enzyme-labeled second physiologically active substance, which specifically binds to the sample substance to be measured is dropped on the dropping portion to cause a reaction, and the reaction solution is filled in the longitudinal direction of the filter.
  • the washing solution is dropped, and then the amount of the sample substance is measured by measuring the amount of enzyme activity on the fibrous porous film.
  • the ratio of a sample containing a sample substance to a substance in which a ligand is bound to a physiologically active substance that specifically binds to the sample substance is 20: It should be measured in the range of 1 to 1: 1, preferably in the range of 10: 1 to 2: 1, and more preferably in the range of 10: 1 to 4: 1.
  • reaction solution After a binding reaction in a solution between a substance in which a ligand is bound to a first physiologically active substance that specifically binds to a measurement sample substance and a sample containing the measurement sample substance, the reaction solution is appropriately cooled. After diluting with a diluent, take a certain amount and drop it on the solid support.
  • a glass fiber filter as the porous filter.
  • biotin as the ligand
  • an anti-biotin antibody or streptavidin as the ligand scavenger bound to the porous filter.
  • the ligand capturing agent may be bound to the porous filter via a spacer.
  • an antibody to the ligand that is, an anti-biotin antibody or streptavidin
  • an anti-biotin antibody is overlaid on an antibody against the ligand capture agent, that is, an antibody against the anti-biotin antibody.
  • an anti-streptavidin antibody may be used as a spacer, or an antistreptavidin antibody may be overlaid with an anti-streptavidin antibody.
  • a conjugate in which a ligand is bound to a physiologically active substance that specifically binds to a substance to be measured such as a ligand-labeled antigen or a ligand-labeled antibody
  • a porous filter as a solid phase carrier
  • a sample solution containing the sample substance to be measured are allowed to react in advance in the solution, and then this reaction solution is directly dropped onto the solid phase of the porous filter to which the ligand-capturing agent binds, and then dropped.
  • a second physiologically active substance that specifically binds to the sample substance after the reaction solution that has penetrated in the longitudinal direction of the filter is labeled with an enzyme, for example, an enzyme-labeled antibody or an enzyme-labeled antigen.
  • an enzyme for example, an enzyme-labeled antibody or an enzyme-labeled antigen.
  • the sample solution containing the measurement sample substance and the conjugate in which the ligand is bound to the first physiologically active substance that specifically binds to the measurement sample substance have a volume ratio of 2%.
  • the ratio in the range of 0: 1 to 1: 1, preferably in the range of 10: 1 to 2: 1, for example, in a short time of 15 minutes or less, a very small amount of ng / mL can be measured. Is now possible.
  • the solid-phase carrier has excellent liquid permeability, Absorbent layer made of glass fiber that has excellent ability to physically or chemically immobilize proteins and glycoproteins and has moderate physical strength, and under which cellulose for absorbing the solution that has passed through the solid phase carrier It is preferable to use a reaction vessel (ie, a plotting device) for immunological measurement in which the above is combined.
  • the physical immobilization method of the spacer or the ligand capture agent on the solid phase carrier is as follows. From the top of the solid phase carrier, sequentially apply a wetting buffer and biotin Add a protein solution (ligand capture agent solution) that recognizes the target, a blocking solution containing a blocking solution containing a blocking solution to prevent nonspecific adsorption of contaminants other than the target substance, and freeze-dry.
  • a method for preparing an immobilized solid phase carrier can be used.
  • a glass fiber carrier is used, for example, according to the method of Ishikawa et al. (Enzyme immunoassay: Medical Shoin, p94, 1978), aminoalkylsilane-glutaraldehyde.
  • a plotting device is created by combining with the absorption layer, the same solution as in the physical immobilization method described above is added in the same order, and freeze-dried to obtain the solid phase on which the ligand capturing agent is immobilized. Can be made.
  • an anti-biotin antibody or streptavidin can be used as a ligand capturing agent.
  • the following probe is used. Higher sensitivity can be obtained when immobilization is performed through one.
  • the anti-biotin antibody is immobilized on the solid support via an antibody against the anti-biotin antibody
  • streptavidin is immobilized on the solid support via the anti-streptavidin antibody
  • the anti-streptavidin antibody A solid phase prepared by immobilizing streptavidin on a solid phase in which an anti-streptavidin antibody is overlaid on an antibody against the above can further increase the advantage of the assay method of the present invention.
  • a solid phase immobilized with a ligand-capturing agent can be used effectively in order to measure a wide variety of substances to be measured quickly, with high sensitivity, specificity, and high accuracy in an immunological measurement method.
  • biotin As a ligand of a ligand-labeled antigen or a ligand-labeled antibody that immunologically specifically reacts with a substance to be measured (a substance to be measured) in a sample. Since a biotin-labeled antigen or a biotin-labeled antibody reacts quickly with a substance to be measured without impairing its own immunological activity, measurement time can be shortened. If the activity of the antigen or antibody is changed due to biotin labeling, change the side chain length of the biotinylation reagent or use a method of labeling the terminal sugar chain of the antigen or antibody with biotin after oxidation. Can be.
  • the volume ratio of the solution of the physiologically active substance to be measured to the solution containing the ligand label is changed according to the measured sensitivity or the reaction between the target substance and the ligand label, and the concentration of the ligand label substance is increased.
  • the volume ratio is such that the concentration of the substance to be measured is maintained, that is, the ligand-labeled solution: the solution of the substance to be measured is allowed to react in the range of 1:20 to 1: 1, preferably 1:10 to 1: 2.
  • the measurement sample can be diluted with an appropriate diluent and then mixed with the ligand-labeled product. It is also possible to dilute and then use that fixed amount as the sample volume.
  • a measurement sample substance is reacted with a ligand-labeled antigen or a ligand-labeled antibody, and an enzyme-labeled antibody in a solution in advance, and the reaction mixture is added to a solid phase on which a ligand capture agent is immobilized. After removing the excess enzyme-labeled antibody by adding the enzyme, an enzyme substrate may be further reacted.
  • the sample substance to be measured includes allergen-specific IgE antibodies, non-specific IgE, hepatitis virus (A, B, C, D, E), and viral infection.
  • IgE antibodies include allergen-specific IgE antibodies, non-specific IgE, hepatitis virus (A, B, C, D, E), and viral infection.
  • HIV HIV, HTLV, herpes, oral, rubella etc.
  • protozoal infections toxoplasma, spirochetes, etc.
  • fungal infections chlorlamydia, Candida, Tricomonas etc.
  • various infectious disease markers CEA, AFP , PSA, CA19-9, CA125, ferritin
  • tumor markers such as DUPAN II
  • inflammatory markers such as CRP, AS ⁇ , RF, etc.
  • myocardial infarction markers such as troponin I, troponin T, myoglobin And the like.
  • a blot device incorporating a glass filter and an absorption layer was used to add 1 OmM phosphate from the top of the filter at room temperature.
  • Physiological saline buffer wetting solution: pH 7.4
  • 1 to 1 000 g / mL anti-biotin antibody goat IgG solution (1 o OmM citrate buffer: pH 3 to 4, or 1 OmM phosphate mono-saline buffer: pH 7.4) was added, and the mixture was allowed to stand for 10 to 30 minutes.
  • a blocking agent and a preservative containing Block Ace (Dainippon Pharmaceutical Co., Ltd.) were added.
  • the blotting apparatus was freeze-dried for 24 hours to prepare an anti-biotin antibody-immobilized solid phase.
  • TMBZ tetramethylbenzidine
  • the reflectance ( ⁇ / S) of the blue color tone on the surface of the solid support was measured with an integrating sphere detector.
  • the initial rate of the enzymatic reaction was measured within 6 minutes after dropping the liquid into the cutting device.
  • the reflectance a and b seconds after the addition of the substrate is (K / S) a and (K / S) b, respectively, the initial velocity ( ⁇ / S) is expressed by the following equation.
  • ⁇ / S ((K / S) b ⁇ (K / S) a) ⁇ 60 / (ba)
  • unsensitized glass fiber was used.
  • 10 mM phosphate-saline buffer (wet solution: pH 7.4) at room temperature from the top of the filter, and then add 1 to 100 ⁇ g / mL Human IgE monoclonal antibody solution (10 OmM citrate buffer: pH 3-4, or 1 OmM phosphate mono-saline buffer: pH 7.4) Add 50 / L and leave for 10-30 minutes.
  • a blocking agent containing block ace and a stabilizer mixture containing preservatives were added.
  • the blotting apparatus was freeze-dried for 24 hours to prepare an anti-human IgE monoclonal antibody-immobilized solid phase.
  • the diluent and the sample were mixed in a ratio of 1: 1.
  • 50 L as the sample, the same operation as in the method of the present invention described above was performed and a human IgE sample was prepared. Was measured, and the measured value was determined (Comparative Example 1).
  • Table 1 shows the measurement results. Table 1 Comparison of sensitivity of standard human IgE antibody concentration Sample Method of the present invention Comparative Example 1 Method of the present invention Z Comparative Example
  • a total of 15 block devices for 5 ⁇ 3 persons incorporating the anti-biotin antibody-immobilized solid phase prepared in the above (1) and 2) were prepared. From the top of each, add 50 L each of a blocking solution containing a block ace and a blocking solution containing a preservative, and then add a biotinylated house dust 2 allergen solution (H 2), a biotinylated mite Dermatophagoides mite 'allergen Solution (D2), Piotinylated Camellia pollen 'Allergen solution (G3)', Piotinylated bush pollen 'Allergen solution (W1), Piotinylated alternaria' Allergen solution (M6) ⁇ L was added and the mixture was heated at 37 ° C for 5 minutes.
  • H 2 biotinylated house dust 2 allergen solution
  • D2 biotinylated mite Dermatophagoides mite 'allergen Solution
  • G3 Piotinylated Camellia pollen
  • each of the blotters was coated with a horseradish peroxidase-labeled anti-human IgE mouse monoclonal antibody solution for 20 ⁇ l each.
  • Each L was added, and the mixture was heated at 37 ° C for 2 minutes.
  • Each block was supplied twice with 80 L of a wash solution consisting of buffered saline containing 0.5% Tween-20, after the previously supplied solution was completely absorbed, and applied to the filter and solid phase.
  • a 10 mM phosphate-saline solution at room temperature was applied from the upper part of the filter to a blotting device incorporating a glass filter and an absorption layer.
  • Buffer solution wetting solution: pH 7.4
  • Add 50 zm Caro then 1 to 5 zg / mL allergen solution 10 OmM citrate buffer: pH 3 to 4 or 1 OmM phosphate-saline buffer (Wetting liquid: pH 7.4)
  • Add 100 / m leave for 10 to 30 minutes, then add 5 O ⁇ L of blocking ace-containing blocking agent and preservative-containing stabilizer mixture did.
  • the block device was freeze-dried all day and night to obtain an allergen-immobilized solid phase.
  • TMBZ tetramethylbenzidine
  • a wetting solution to the block device containing a glass filter from the top of the filter at room temperature, and then add a 1 to 1000 ⁇ g / mL streptavidin solution (10 OmM citrate buffer: pH 3 to 4, Alternatively, 10 mM phosphate-saline buffer: pH 7.4) was added, and the mixture was allowed to stand for 10 to 30 minutes, and then a mixed solution of a blocking agent and a stabilizing agent was added. It was freeze-dried all day and night to prepare a streptavidin-immobilized solid phase.
  • streptavidin solution 10 OmM citrate buffer: pH 3 to 4, Alternatively, 10 mM phosphate-saline buffer: pH 7.4
  • Example 2 As a comparative example, the same method as in Example 1 was used, using Comparative Example 2 (in which the allergen was directly bound to the solid phase) described in Example 1 as it was, and using the solid phase of 2) above.
  • the allergen-specific IgE of the patient serum was measured by the method, almost the same value as in Example 1 was obtained, and the sensitivity was about twice as high. That is, it was confirmed that the same result was obtained even when the solid phase was changed to streptavidin.
  • a glass fill punched to a diameter of 8 mm was immersed in a 2% solution of 3-aminopropylethoxysilane-aceton at 45 ° C for 24 hours, washed with acetone and dried. Dried. Then, this was immersed in a 1% aqueous solution of glutaraldehyde (GA) for 1 hour at room temperature, and washed with 0.25M phosphate buffer (pH 7.4) until the smell of glutaraldehyde disappeared.
  • a glutaraldehyde activating filter was prepared.
  • Example 1 Using the solid phase obtained above, the detection sensitivity was determined by the same method as in Example 1. As a result, it was confirmed that the same results as in Example 1 were obtained.
  • test sample solution 10 volumes were used for 1 volume of the biotinylated allergen solution.
  • 5 parts of each of these reaction mixtures was added for 2 minutes. Warmed at ° C. After heating, 20 ⁇ L of a horseradish peroxidase-labeled anti-human IgE mouse monoclonal antibody solution was added from the top of each block device and heated at 37 ° C. for 2 minutes.
  • G 1 100> 100> 100> 100> 47.5 33.3 1.5 2.4 33.3 37.7
  • G6 100> 100> 100> 100> 30.9 35.2 1.6 3.3 10.7 34.8
  • the porous solid phase was incorporated into a plotting device.
  • Each plotting device was washed with a washing solution consisting of a buffered saline solution containing 0.5% Tween-20 and 80 j. L 2 times after the previously supplied solution has been completely absorbed and supplied. The remaining excess labeled antibody was removed.
  • TMB Z blue color tone
  • a 1 OmM phosphate / saline buffer solution (wetting solution: pH 7.4) was used at room temperature from the top of the filter using a plotting device assembled with an untreated glass fiber filter. ) And then add 100 ⁇ g / mL anti-human CA 19-9 monoclonal antibody solution (10 OmM citrate buffer: pH 3-4, or 10 mM phosphate mono-saline buffer: p After adding H7.4), the mixture was allowed to stand for 10 to 30 minutes, and then a blocking agent containing Block Ace, a preservative, and a stabilizer mixed solution containing a preservative were added.
  • the plotting apparatus was used to prepare a freeze-dried anti-human C A19-9 monoclonal antibody-immobilized solid phase all day and night. This solid phase was used as a comparative example.
  • the measurement method was as follows: the diluted solution was mixed with the CA 19-9 standard solution at a ratio of 1: 1, and 50 ⁇ L was used as a sample. Then, measured values were obtained. Table 10 also shows the results. Table 10 Measured values of standard solution of CA 19-9
  • a single ligand can be prepared using an immunoassay without preparing and installing separate solid phases corresponding to the substance to be measured.
  • a capture agent-immobilized solid phase it is possible to measure various types of target substances simply, quickly, with high sensitivity, and with high accuracy.

Abstract

L'invention concerne un procédé permettant de mesurer une substance échantillon physiologiquement active avec rapidité et avec une sensibilité élevée. Ce procédé consiste à amener une solution de cette substance échantillon à mesurer au contact d'une solution d'une substance constituée par une première substance physiologiquement active se liant spécifiquement à cette substance échantillon et un ligand lié à ladite substance dans un rapport volumique compris entre 1:1 et 20:1, d'où la formation d'un complexe de cette substance physiologiquement active liée au ligand et à la substance échantillon à mesurer dans la solution. Ledit procédé consiste ensuite à verser la solution contenant ce complexe dans un filtre poreux auquel un agent de capture dudit ligand est lié, d'où la liaison de la fraction de ligand du complexe audit agent de capture, à verser une solution d'une seconde substance physiologiquement active marquée par une enzyme se liant spécifiquement à la substance échantillon à mesurer dans le filtre afin de lier cette substance physiologiquement active à marquage enzymatique au complexe de la première substance physiologiquement active et à la substance échantillon à mesurer liée au filtre via l'agent de capture et la fraction de ligand, à nettoyer le filtre pour éliminer la seconde substance physiologiquement active non liée à marquage enzymatique, puis à mesurer l'activité de l'enzyme liée au filtre.
PCT/JP2001/001196 2000-02-23 2001-02-20 Procede de mesure d'une substance echantillon physiologiquement active au moyen d'un filtre poreux WO2001063283A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
KR1020027010881A KR20020086558A (ko) 2000-02-23 2001-02-20 다공성 필터를 이용하는 생리활성 시료물질의 측정방법
AU2001232350A AU2001232350A1 (en) 2000-02-23 2001-02-20 Method of measuring physiologically active sample substance by using porous filter

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2000/45380 2000-02-23
JP2000045380A JP4545869B2 (ja) 2000-02-23 2000-02-23 多孔性フィルタを用いる生理活性試料物質の測定方法

Publications (1)

Publication Number Publication Date
WO2001063283A1 true WO2001063283A1 (fr) 2001-08-30

Family

ID=18567975

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2001/001196 WO2001063283A1 (fr) 2000-02-23 2001-02-20 Procede de mesure d'une substance echantillon physiologiquement active au moyen d'un filtre poreux

Country Status (5)

Country Link
JP (1) JP4545869B2 (fr)
KR (1) KR20020086558A (fr)
AU (1) AU2001232350A1 (fr)
TW (1) TWI275796B (fr)
WO (1) WO2001063283A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2108656A1 (fr) 2008-03-19 2009-10-14 Beninati, Concetta Fragments de protéines antigéniques de streptococcus pneumoniae
US7790187B2 (en) 2005-03-08 2010-09-07 Kenton S.R.L. Chimeric recombinant antigens of Toxoplasma gondii

Families Citing this family (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP4739755B2 (ja) * 2002-10-11 2011-08-03 ゼットビーエックス コーポレーション 診断装置
KR20070012419A (ko) 2004-04-21 2007-01-25 가부시키가이샤 에이앤드티 면역학적 정량법과 시약
JP2007121095A (ja) * 2005-10-27 2007-05-17 Toyobo Co Ltd 非特異反応が抑制されるように改良された測定容器
JP4904182B2 (ja) * 2007-03-19 2012-03-28 財団法人電力中央研究所 カドミウム、鉛、又は３価クロムの免疫学的定量方法及びそれらの免疫学的定量装置、並びに、それらに用いるカドミウム、鉛、又は３価クロム錯体固定化膜
JP5118479B2 (ja) * 2007-12-28 2013-01-16 田中貴金属工業株式会社 イムノクロマト分析法及びイムノクロマト分析キット
JPWO2010041595A1 (ja) * 2008-10-08 2012-03-08 東洋紡績株式会社 生理活性物質測定システム
JP5716501B2 (ja) * 2011-04-04 2015-05-13 東洋紡株式会社 検体の前処理方法
JP6531307B2 (ja) * 2015-04-08 2019-06-19 株式会社パートナーファーム 固相反応容器及びこれを用いた測定方法
JP7121760B2 (ja) 2018-02-02 2022-08-18 日本ケミファ株式会社 生化学反応用基体及び分析装置
WO2021025066A1 (fr) 2019-08-07 2021-02-11 株式会社パートナーファーム Récipient à réaction en phase solide et procédé de mesure dans lequel ce dernier est utilisé
WO2021095858A1 (fr) * 2019-11-14 2021-05-20 インターメディック株式会社 Procédé de mesure de substance physiologiquement active

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS60500731A (ja) * 1983-04-08 1985-05-16 クイデル ヒト絨毛性性腺刺激ホルモンの検出
JPS6366462A (ja) * 1986-06-20 1988-03-25 ベ−リンガ−・マンハイム・ゲゼルシヤフト・ミツト・ベシユレンクテル・ハフツング 不均一系免疫分析用の免疫反応性多孔性担体材料及びその製法
JPH01312463A (ja) * 1988-06-10 1989-12-18 Dai Ichi Pure Chem Co Ltd 免疫学的測定法
JPH02145967A (ja) * 1988-04-07 1990-06-05 Bio Rad Lab Inc ビオチン架橋および万能固相を利用するイムノアッセイ

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4632901A (en) * 1984-05-11 1986-12-30 Hybritech Incorporated Method and apparatus for immunoassays
JP3134231B2 (ja) * 1991-04-16 2001-02-13 東洋紡績株式会社 固相生物学的特異反応測定法およびそのための器具
JPH10253632A (ja) * 1997-03-10 1998-09-25 Nissui Pharm Co Ltd 分析方法、キット及び装置
JPH11201969A (ja) * 1998-01-14 1999-07-30 Wakamoto Pharmaceut Co Ltd 簡易遊離ヘモグロビン測定方法および測定用キット
JPH11326326A (ja) * 1998-05-13 1999-11-26 Dainippon Printing Co Ltd 免疫検査体
JP2000088851A (ja) * 1998-09-10 2000-03-31 Internatl Reagents Corp 生物学的特異反応測定法及び装置

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS60500731A (ja) * 1983-04-08 1985-05-16 クイデル ヒト絨毛性性腺刺激ホルモンの検出
JPS6366462A (ja) * 1986-06-20 1988-03-25 ベ−リンガ−・マンハイム・ゲゼルシヤフト・ミツト・ベシユレンクテル・ハフツング 不均一系免疫分析用の免疫反応性多孔性担体材料及びその製法
JPH02145967A (ja) * 1988-04-07 1990-06-05 Bio Rad Lab Inc ビオチン架橋および万能固相を利用するイムノアッセイ
JPH01312463A (ja) * 1988-06-10 1989-12-18 Dai Ichi Pure Chem Co Ltd 免疫学的測定法

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7790187B2 (en) 2005-03-08 2010-09-07 Kenton S.R.L. Chimeric recombinant antigens of Toxoplasma gondii
US7867503B2 (en) 2005-03-08 2011-01-11 Sigma-Tau Industrie Farmaceutiche Riunite S.P.A. Chimeric recombinant antigens of Toxoplasma gondii
EP2108656A1 (fr) 2008-03-19 2009-10-14 Beninati, Concetta Fragments de protéines antigéniques de streptococcus pneumoniae

Also Published As

Publication number Publication date
JP2001235471A (ja) 2001-08-31
TWI275796B (en) 2007-03-11
JP4545869B2 (ja) 2010-09-15
AU2001232350A1 (en) 2001-09-03
KR20020086558A (ko) 2002-11-18

Similar Documents

Publication Publication Date Title
US5081013A (en) Immunodiagnostic device and method
US20110003310A1 (en) Oral fluid rapid immunochromatography test
JP3334558B2 (ja) 酵素免疫測定方法及び試験片
JP4209978B2 (ja) ポリエチレングリコール誘導化生体分子及び不均質検出法におけるその使用
JP4579414B2 (ja) リガンド結合アッセイおよび阻害性分析物分離領域を有するキット
JP2504923B2 (ja) 免疫学的測定方法
JPH0566547B2 (fr)
JPH08501387A (ja) ボード上に陰性および陽性対照を有する分析試験装置
CN111164095A (zh) 用于改进分析物检测的测定方法
JPH01113662A (ja) 特異的結合アッセイ装置および方法
WO2001063283A1 (fr) Procede de mesure d'une substance echantillon physiologiquement active au moyen d'un filtre poreux
JPH0346561A (ja) 分析物検出のための方法、試薬及び試験ストリップ
JPH08501018A (ja) 分析試験装置用のケーシング手段
WO1995035502A1 (fr) Dispositif de detection de substances polyvalentes
JPH1164336A (ja) 免疫クロマトグラフィーによる分析対象物の検出方法
EP1938102A2 (fr) Détection d'un ligand cible améliorée
JP3237848B2 (ja) 試験要素の作製方法
JPH05503148A (ja) 分析装置および方法
KR960016717B1 (ko) 검체의 검출방법 및 시약
JPH11153600A (ja) 免疫測定用試験片及び該試験片を用いる測定法
JP4627607B2 (ja) 複数項目及び総含有量の同時分析可能なイムノクロマトグラフ法及びイムノクロマトグラフ用ストリップ
JP4531252B2 (ja) 粒子を使用した分析法および本方法を行うための試験キット
JPH11133023A (ja) 尿試料を使用する免疫クロマトグラフィー検定の尿素の影響を減らすための配合物
JP4437211B2 (ja) 免疫クロマトグラフィーを用いた測定方法およびそれに用いる検体分析用具
JP2505658B2 (ja) サンプル流体成分の分析測定用テストキャリヤ

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 BY BZ CA CH CN CR CU CZ DE DK DM DZ EE ES FI GB GD GE GH GM HR HU ID IL IN IS KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NO NZ PL PT RO RU SD SE SG SI SK SL TJ TM TR TT TZ UA UG US UZ VN YU ZA ZW

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): GH GM KE LS MW MZ SD SL SZ TZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE TR BF BJ CF CG CI CM GA GN GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
WWE Wipo information: entry into national phase

Ref document number: 1020027010881

Country of ref document: KR

WWP Wipo information: published in national office

Ref document number: 1020027010881

Country of ref document: KR

REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

122 Ep: pct application non-entry in european phase