WO2010095469A1 - イムノクロマト法によるアレルゲン検出方法 - Google Patents
イムノクロマト法によるアレルゲン検出方法 Download PDFInfo
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- WO2010095469A1 WO2010095469A1 PCT/JP2010/001209 JP2010001209W WO2010095469A1 WO 2010095469 A1 WO2010095469 A1 WO 2010095469A1 JP 2010001209 W JP2010001209 W JP 2010001209W WO 2010095469 A1 WO2010095469 A1 WO 2010095469A1
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- Prior art keywords
- allergen
- allergens
- immunochromatography
- denatured
- colloidal gold
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- 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
- G01N33/54366—Apparatus specially adapted for solid-phase testing
- G01N33/54386—Analytical elements
- G01N33/54387—Immunochromatographic test strips
- G01N33/54388—Immunochromatographic test strips based on lateral flow
-
- 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
- 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
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- 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
- G01N33/02—Food
-
- 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
- G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/24—Immunology or allergic disorders
Definitions
- each allergen is extracted from a test sample such as food containing various allergens using an anionic surfactant and thiosulfate, and each allergen is in any denatured / undenatured state. Efficient extraction with an anionic surfactant and thiosulfate, or an anionic surfactant and a nonionic surfactant, and further suppresses non-specific reactions associated with the collapse of colloidal gold bound to the antibody.
- the present invention relates to a method for detecting an allergen by an immunochromatography method that can be detected at once, and a detection kit for an allergen for immunochromatography that can be used therefor.
- an immunoassay method for detecting a target substance comprising a specific antigen or antibody using a specific reaction by an antigen-antibody, a target substance in a sample
- An agglutination method that binds an antibody or antigen sensitized to microparticles by an immune reaction and measures the aggregation state of the microparticles resulting from the binding is a simple immunoassay method, and is generally used because it can be visually judged. It is the method that has been.
- a radioimmunoassay method or enzyme immunity method in which an antibody or antigen labeled with a labeling substance composed of a radioisotope, an enzyme or a fluorescent substance is bound to an analyte in a sample by an immune reaction, and the bound labeling substance is measured.
- Measurement methods or fluorescence immunoassays are also employed. In these immunoassays, competitive reaction and sandwich reaction are widely used. Of these, immunochromatography is known as a so-called sandwich-type reaction measurement method (see, for example, Patent Document 1).
- sandwich-type reaction measurement method see, for example, Patent Document 1
- Allergen detection kits are sold.
- Samples applied to such immunochromatography include biological samples and extracts from foods, but depending on the type of sample, a so-called non-specific reaction that shows a light color at the capture site despite the absence of the specimen. May cause a decrease in accuracy in inspection. Therefore, the buffer contains a polymer having a phosphorylcholine group at a concentration of 0.005 to 0.3 w / v%, and the number average molecular weight of the polymer is 40,000 or more.
- Patent Document 2 that prevents non-specific aggregation and non-specific reaction during measurement and thus enables measurement with high accuracy.
- Applicant can extract allergens quickly and accurately by suppressing non-specific reactions associated with colloidal collapse of gold colloids, even when extracted using an extraction solution containing a denaturant and a reducing agent and adapted to immunochromatography.
- the immunochromatography method which can be performed is proposed (for example, refer patent document 3).
- allergens were sufficiently extracted from a heated test sample and further tested by a simple immunochromatography method, accuracy and simplicity could be dramatically improved.
- the reducing agent used (2-mercaptoethanol) has a unique odor and 2-mercaptoethanol has been designated as a toxic substance on July 1, 2008. Has become difficult. Therefore, there has been a demand for a safer and more efficient extraction method and an immunochromatography method that can be accurately tested without any non-specific reaction when these are applied to the immunochromatography method.
- An object of the present invention is to use an anionic surfactant and a thiosulfate or an anionic surfactant and a nonionic surfactant from a test sample such as a food containing each allergen. , And efficiently extract with an anionic surfactant and thiosulfate, or an anionic surfactant and a nonionic surfactant, regardless of whether each allergen is in a denatured / undenatured state. Furthermore, we offer a method for detecting allergens by immunochromatography that can quickly and accurately detect allergens by suppressing the nonspecific reaction associated with the collapsing of colloidal gold bound to the antibody, and an immunochromatographic allergen detection kit that can be used therefor. There is to do.
- each allergen is in any state of denaturation / non-denaturation without using 2-mercaptoethanol, which has been considered indispensable to sufficiently extract each allergen from a heated test sample. Even in an immunochromatography method that can be efficiently extracted with an anionic surfactant and thiosulfate, or an anionic surfactant and a nonionic surfactant, It has been found that the use of a developing solution containing (FBS: fetal bovine ⁇ ⁇ serum) can solve the above problems, and the present invention has been completed.
- FBS fetal bovine ⁇ ⁇ serum
- the present invention includes (1) a colloidal gold-labeled antibody in which colloidal gold is bound to a monoclonal antibody against a denatured and native allergen, and a monoclonal antibody against a denatured and native allergen that recognizes an epitope different from the colloidal gold labeled antibody.
- the fetal bovine serum (FBS) is developed using a developing solution containing measurement samples of denatured and undenatured allergens extracted using a developing support. ) Is used in an immunochromatographic method characterized by using a developing solution containing at least 10% by weight.
- An allergen detection method (2) an allergen detection method by immunochromatography according to (1), wherein a developing solution containing at least 30% by weight of fetal bovine serum (FBS) is used, and (3 (2) an allergen detection method by immunochromatography as described in (2) above, wherein a developing solution containing at least 50% by weight of fetal bovine serum (FBS), or (4) against denatured and undenatured allergens
- Monoclonal antibody has molecular weight of ⁇ s1 casein as a major component of milk allergen, ⁇ -lactoglobulin as a major component of whey allergen, ovalbumin and ovomucoid as an egg white allergen, gliadin as a major component of wheat allergen, buckwheat major protein 24 kDa and 76 kDa proteins, mainly peanuts
- the present invention also provides (7) a colloidal gold-labeled antibody carrier comprising a colloidal gold-conjugated antibody in which a colloidal gold antibody is bound to a monoclonal antibody against a denatured and undenatured allergen, and a denaturation that recognizes an epitope different from the colloidal gold-labeled antibody.
- an anionic surfactant for extracting the modified and native allergen from a test sample such as food containing the allergen, From a test sample such as a thiosulfate or a buffer solution containing an anionic surfactant and a nonionic surfactant and a food containing an allergen, an anionic surfactant and a thiosulfate, or
- An immunochromatographic allergen detection kit comprising a sample carrier that can be obtained, and fetal bovine serum (FBS) or a developing solution containing fetal bovine serum (FBS), and (8) against denatured and undenatured allergens
- Monoclonal antibody has molecular weight of ⁇ s1 casein as a major component of milk allergen, ⁇ -lactoglobulin as a major component of whey allergen, ovalbumin and
- an allergen by the immunochromatography method of the present invention even when an anionic surfactant and thiosulfate, or an anionic surfactant and a nonionic surfactant are used, gold bound to the antibody is used. Non-specific reactions associated with colloid breakdown can be suppressed, and various allergens can be detected quickly and accurately.
- the method for detecting an allergen by the immunochromatography method of the present invention includes a colloidal gold-labeled antibody in which a colloidal gold antibody is bound to a monoclonal antibody against a denatured and undenatured allergen, and a denatured and undenatured antibody that recognizes a different epitope from the gold colloid-labeled antibody.
- a developing solution containing at least 10% by weight of fetal bovine serum (FBS) The method is not particularly limited, but a developing solution containing 20 to 100% by weight of fetal bovine serum (FBS) is preferably used, and a developing solution containing 30 to 100% by weight is more preferable. It is particularly preferable to use a developing solution containing 40 to 100% by weight, and it is even more preferable to use a developing solution containing 50 to 100% by weight.
- the immunochromatographic allergen detection kit of the present invention includes a colloidal gold-labeled antibody carrying a colloidal gold antibody bound to a monoclonal antibody against denatured and undenatured allergens, an epitope different from the colloidal gold-labeled antibody.
- the fetal bovine serum (FBS) concentration in the developing solution is less than 10% by weight, a nonspecific reaction tends to occur, which is not preferable.
- various additives such as other surfactants, preservatives, and inorganic salts are suspended, emulsified or dissolved in the buffer solution as necessary.
- the pH of the buffer is preferably 4 to 10, and particularly preferably 6 to 8.
- a phosphate buffer (PBS) or a Tris buffer can be suitably exemplified.
- a method for producing a colloidal gold-labeled antibody in which colloidal gold is bound to the above monoclonal antibody is not particularly limited, including a conventionally known method.
- a 2 mM boronic acid solution is added to a colloidal gold solution prepared to pH 9.0 with 0.2 M potassium carbonate solution.
- An example is a method in which a solution in which a monoclonal antibody is dissolved in an acid buffer (pH 9.0) is added and reacted at room temperature for 30 minutes, and then a 10% BSA solution is added, further reacted for 15 minutes, and centrifuged.
- the colloidal gold-labeled antibody carrier can be produced by applying the colloidal gold-labeled antibody produced above onto, for example, a glass wool conjugate pad and drying it.
- the development support is subjected to a blocking treatment after a buffer solution containing a monoclonal antibody against a modified and unmodified allergen that recognizes an epitope different from that of a colloidal gold-labeled antibody, for example, is linearly applied to a nitrocellulose membrane and dried. Can be produced.
- anionic surfactants in the buffer used when preparing a measurement sample by extracting denatured and undenatured allergens higher alcohol sulfate, alkyl naphthalene sulfonate, alkyl benzene sulfonate, alkyl A phosphoric acid ester salt etc. can be mentioned, Specifically, sodium dodecyl sulfate (SDS) can be illustrated suitably.
- SDS sodium dodecyl sulfate
- the thiosulfate include sodium thiosulfate, potassium thiosulfate, and ammonium thiosulfate.
- sodium thiosulfate can be preferably exemplified.
- Nonionic surfactants include polyoxyethylene alkyl ether, polyoxyethylene alkyl phenyl ether, polyoxyethylene fatty acid ester, polyglycerin fatty acid ester, polyoxyethylene fatty acid amide, polyoxyethylene alkylamine, sorbitan fatty acid ester, etc.
- polyoxyethylene (20) sorbitan monolaurate (Tween 20) can be preferably exemplified.
- the concentration of the anionic surfactant is 0.1 to 2.0%, preferably 0.25% to 0.5%, and the concentration of thiosulfate is 0.1 to 5.0%, preferably 0.1% to 1.0%, and the concentration of the nonionic surfactant is 0.01 to 1.0%, preferably 0.05 to 0.2%. It is preferable to use a buffer solution containing an ionic surfactant and thiosulfate, or an anionic surfactant and a nonionic surfactant, because extraction efficiency is high and nonspecific reaction can be suppressed.
- sample carrier capable of supporting the measurement sample
- a sample pad made of glass wool can be exemplified.
- the sample carrier, the colloidal gold-labeled antibody support, the development support, and preferably the other end of the development support are connected to the other end of the development support, such as an absorbent pad, for immunochromatography. It can be a test piece.
- the allergen in the measurement sample moves due to capillary action or the like and binds to the colloidal gold labeled antibody, and this antigen-antibody complex is
- the antigen-antibody complex is captured at a predetermined position where a monoclonal antibody against a modified and native allergen that recognizes an epitope different from the colloidal gold-labeled antibody is still moved by capillary action or the like on the development support, Allergens can be detected by the presence or absence of a colored line appearing at a predetermined position.
- Monoclonal antibodies against the above-mentioned modified and native allergens include ⁇ s1 casein as a major component of milk allergen, ⁇ -lactoglobulin as a major component of whey allergen, ovalbumin and ovomucoid as an egg white allergen, as a major component of wheat allergen
- Two types of monoclonal antibodies specifically recognizing denatured and native allergens selected from gliadin, proteins of buckwheat major proteins of 24 kDa and 76 kDa, and arachi, a major protein of peanuts can be preferably exemplified. .
- anti- ⁇ s1 casein monoclonal antibody Pas1CN1 produced by hybridoma (FERM-BP-10263) and hybridoma (FERM-BP-10264) are produced as anti- ⁇ s1 casein monoclonal antibodies prepared by the present inventors.
- Examples of the anti- ⁇ s1 casein monoclonal antibody PasCN2 can be mentioned.
- Examples of the anti- ⁇ -lactoglobulin monoclonal antibody include an anti- ⁇ -lactoglobulin monoclonal antibody P ⁇ LG3 produced by a hybridoma (FERM-ABP-11237) and a hybridoma (FERM-ABP-11238).
- Anti- ⁇ -lactoglobulin monoclonal antibody P ⁇ LG4 produced by The hybridoma (FERM-BP-10263) and the hybridoma (FERM-BP-10264) were incorporated on February 24, 2005 by the National Institute of Advanced Industrial Science and Technology (AIST) Tsukuba City East 1-1 1)
- the hybridoma (FERM-ABP-11237) and hybridoma (FERM-ABP-11238) were commissioned on February 22, 2010 by the National Institute of Advanced Industrial Science and Technology (AIST). Address: Tsukuba, Ibaraki Prefecture 1-1-1 Higashi Tsukuba Center Central 6)
- anti-ovalbumin monoclonal antibody PDOA3 produced by a hybridoma (FERM-ABP-11235) and anti-ovalbumin monoclonal antibody PDOA4 produced by a hybridoma (FERM-ABP-11236) can be exemplified.
- Anti-ovomucoid monoclonal antibody PNOM1 produced by hybridoma (FERM-BP-10279), anti-ovomucoid monoclonal antibody PNOM2 produced by hybridoma (FERM-BP-10280), and anti-ovomucoid monoclonal produced by hybridoma (FERM-BP-10277)
- Antibody PDOM1 and anti-ovomucoid monoclonal antibody PDOM produced by hybridoma (FERM-BP-10278) It can be mentioned.
- Hybridoma (FERM-ABP-11235) and Hybridoma (FERM-ABP-11236) were incorporated on February 22, 2010 by the National Institute of Advanced Industrial Science and Technology (AIST) Tsukuba City East 1-1 1)
- the hybridoma (FERM-BP-10279), hybridoma (FERM-BP-10280), hybridoma (FERM-BP-10277), and hybridoma (FERM-BP-10278) It was entrusted to the National Institute of Advanced Industrial Science and Technology Patent Biological Deposit Center (address: Tsukuba City Center 1-1-1 Tsukuba Center Central 6th) on February 24, 1992.
- anti-wheat gliadin monoclonal antibody examples include an anti-wheat gliadin monoclonal antibody PGL1 produced by a hybridoma (FERM-BP-10267) and an anti-wheat gliadin monoclonal antibody PGL2 produced by a hybridoma (FERM-BP-10268).
- Hybridoma (FERM-BP-10267) and Hybridoma (FERM-BP-10268) were incorporated on February 24, 2005 at the National Institute of Advanced Industrial Science and Technology (AIST) Tsukuba City East 1-1 It is entrusted to 1 Tsukuba Center Central 6).
- anti-soba protein monoclonal antibody examples include an anti-24 kDa protein monoclonal antibody PBW5 produced by a hybridoma (FERM-ABP-11241) and an anti-76 kDa protein monoclonal antibody PBW2 produced by a hybridoma (FERM-BP-10273). .
- Hybridoma (FERM-ABP-11241) was received on February 22, 2010 by the National Institute of Advanced Industrial Science and Technology Patent Biological Depositary (Address: 1-1-1 Tsukuba Center, Tsukuba City, Ibaraki) , Hybridoma (FERM-BP-10273) was commissioned on February 24, 2005 to the National Institute of Advanced Industrial Science and Technology, Patent Biological Deposit Center (address: 1-1-1 Tsukuba Center, Tsukuba City, Ibaraki) Has been. *
- anti-peanut Ara h1 protein monoclonal antibody anti-native Ara11 monoclonal antibody PAh1-5 produced by hybridoma (FERM-ABP-11240) and anti-native Ara h1 protein monoclonal produced by hybridoma (FERM-ABP-11239)
- the antibody PAh1-4 can be mentioned.
- Hybridoma (FERM-ABP-11240) and Hybridoma (FERM-ABP-11239) were incorporated on February 22, 2010 by the National Institute of Advanced Industrial Science and Technology (AIST) Tsukuba City East 1-1 1st Tsukuba Center Center 6)
- Model meat product A meat product was selected as a model food for the quantitative test, and a model meat product containing egg protein at each concentration was prepared according to the formulation shown in Table 1.
- the red pork used was prepared by removing fats and streaks from pork loin and grinding it to 5 mm. Additives were weighed according to each formulation, mixed in a food processor, and filled into a PVC tube.
- Heating temperature / time Heating was performed by heating at 75 ° C. for 30 minutes. After heating, the sample made uniform by a food processor was used as a sample for detection.
- Tween 20 is not present in the extraction with the immunochromatography kit.
- the measurement sample [3] is for examining whether Tween 20 contributes to extraction efficiency because SDS and Tween 20 coexist and are extracted. 4) Confirmation of detection by immunochromatography 20 ⁇ l of the prepared gold colloid-labeled antibody, 30 ⁇ l of fetal bovine serum (FBS) as a developing solution, 50 ⁇ l of measurement sample [1], measurement sample [2] and measurement sample [3] are added, The test was performed on an immunochromatographic strip and the detection was confirmed.
- FBS fetal bovine serum
- the model meat product heated at 75 ° C. for 30 minutes was judged as + up to 2 ppm in all measurement samples, and there was no non-specific reaction at 0 ppm.
- the model meat product heated at 121 ° C. for 20 minutes was determined to be + w up to 2 ppm in the measurement sample [3], and egg protein could be detected up to 2 ppm. There was no non-specific reaction at 0 ppm. From this, in the case of the extraction method combining an anionic surfactant and a nonionic surfactant, the extraction efficiency is high, and there is no non-specific reaction by using FBS in the developing solution, and An accurate immunochromatography kit can be constructed.
- Table 6 shows the detection results. Judgment was expressed in the order of +, + w, +-in order from the strongest line, and negative was set as-.
- the measurement sample [3] was determined to be + w up to 2 ppm, and milk protein could be detected up to 2 ppm. There was no non-specific reaction at 0 ppm. Therefore, the extraction efficiency is the highest in the case of the extraction method combining an anionic surfactant and a nonionic surfactant as in the measurement sample [3], and FBS is used as a developing solution. It was possible to construct an immunochromatography kit quickly and accurately without any non-specific reaction.
- Table 9 shows the detection results. Judgment was expressed in the order of +, + w, +-in order from the strongest line, and negative was set as-.
- a glass wool sample pad for the test solution spot and a glass wool absorption pad for absorbing the test solution are prepared separately.
- the absorbent pads were pasted in this order to form an immunochromatographic strip.
- Wheat protein was prepared from commercially available wheat powder according to the method of Hatakeyama et al.
- the sample for detection was a model meat product having the composition shown in Table 12, and the heating temperature / time and sample pretreatment were performed under the same conditions as described above.
- the sodium thiosulfate concentration was detected up to 2 ppm from 0% to 10.0%, and was judged to be + from 0% to 5.0%, but in particular from 0.1% to 2.0% Up to the highest visibility. There was no non-specific reaction at 0 ppm. From this, in the case of the extraction method combining an anionic surfactant and thiosulfate, the extraction efficiency is high, and there is no non-specific reaction by using FBS in the developing solution, and it is quick and accurate. It became possible to construct an immunochromatography kit.
- Table 17 shows the detection results. Judgment was expressed in the order of +, + w, +-in order from the strongest line, and negative was set as-.
- the model meat product heated at 75 ° C. for 30 minutes was judged as + ⁇ up to 2 ppm in the measurement sample [3], and there was no non-specific reaction at 0 ppm.
- the measured sample [3] was determined to be + w up to 2 ppm, and the buckwheat protein could be detected up to 2 ppm. There was no non-specific reaction at 0 ppm. From this, the extraction method combining an anionic surfactant and a nonionic surfactant has high extraction efficiency, and by using FBS as a developing solution, there is no non-specific reaction and it can be performed quickly.
- an immunochromatography kit with high accuracy can be constructed.
- allergen detection kit for immunochromatography.
Abstract
Description
1.材料及び方法
(1)金コロイド標識抗体の作製
2mMホウ酸緩衝液(pH9.0)で1mg/mlとなるようにPDOA4(FERM-ABP-11236)のMAb溶液を調製した。あらかじめ0.2M炭酸カリウム溶液でpH9.0に調製した金コロイド溶液(シグマ社製)5mlにMAb溶液を500μl加え、室温で30分間反応した後、10%BSA溶液を635μl加え、さらに15分間反応させた。遠心分離を行い、1%BSA溶液でOD525=1.0になるよう調製した。
PBSで4mg/mlとなるようにPDOA3(FERM-ABP-11235)のMAb溶液を調製し、ニトロセルロースメンブレンに直線状に塗布し乾燥させた。その後、1%スキムミルクを含むTBSで37℃、1時間ブロッキング後、TBSで洗浄し乾燥させた。
抗体固定化メンブレンに加えて、被検液スポット用のガラスウール製サンプルパッド、被検液吸収用のガラスウール製吸収パッドを別途用意し、サンプルパッド、抗体固定化メンブレン、吸収パッドの順にそれぞれ貼り付け、イムノクロマトストリップとした。検出用サンプルには、以下のモデル食肉製品を供試した。
穐山ら(特定原材料(卵)測定の厚生労働省通知ELISA法の複数機関による評価研究.食品衛生学雑誌,44, 2003, 213-219)に従い、市販鶏卵より卵タンパク質を調製した。
定量試験のためのモデル食品として食肉製品を選択し、表1に示す配合にて各濃度の卵タンパク質を含むモデル食肉製品を作製した。豚赤肉は、豚ロース肉より脂、スジを除去し、5mmで挽肉にしたものを使用した。各配合に従い添加物を計量し、フードプロセッサーにて混合後、塩ビチューブに充填を行った。
加熱は、75℃30分加熱したものを用意した。加熱後、フードプロセッサーにて均一としたものを検出用サンプルとした。
検出用サンプル1gを量り取り、それに抽出液として、0.5%SDSとチオ硫酸ナトリウムが最終濃度で0%~10.0%含まれるPBSを19ml加え撹拌し、沸騰水中で1時間加熱し、冷却遠心後、上清を測定サンプルとした。
調製した金コロイド標識抗体を20μl、展開液として牛胎児血清(FBS)を30μl、測定サンプルを50μl加え、イムノクロマトストリップに供試し、検出を確認した。
次に検出結果を表2に示す。判定はラインの強い方から順に+、+w、+-と表記し、陰性を-表記とした。
1.材料及び方法
1)以下の「加熱温度・時間」、「サンプルの前処理」及び「イムノクロマトグラフィーによる検出の確認」を除いては、実施例1と同様に行った。
2)加熱温度・時間
加熱は、75℃30分と121℃20分加熱したものを用意した。
3)サンプルの前処理
加熱後、フードプロセッサーにて均一としたものを検出用サンプルとした。検出用サンプル1gを量り取り、それに、0.5%SDSを含むPBSを19ml加え撹拌し、沸騰水中で1時間加熱し、冷却遠心後、上清を測定サンプル〔1〕とした。また、測定サンプル〔1〕に最終濃度が0.2%となるようにTween20を加えたものを測定サンプル〔2〕とした。さらに、0.5%SDS及び0.2%Tween20を含むPBSを19ml加え撹拌し、沸騰水中で1時間加熱し、冷却遠心後、上清を測定サンプル〔3〕とした。上記測定サンプル〔2〕は、SDS抽出後にTween20を加えているため、抽出に関与するのではなく、イムノクロマトキットでサンプルを測定する際に、Tween20が存在していることがイムノクトマトキットの感度に関与するかを検討するためのものであり、上記測定サンプル〔3〕は、SDSとTween20を共存させ抽出しているため、抽出効率にTween20が貢献するかを検討するためのものである。
4)イムノクロマトグラフィーによる検出の確認
調製した金コロイド標識抗体を20μl、展開液として牛胎児血清(FBS)を30μl、測定サンプル〔1〕、測定サンプル〔2〕、測定サンプル〔3〕を50μl加え、イムノクロマトストリップに供試し、検出を確認した。
次に検出結果を表3及び表4に示す。判定はラインの強い方から順に+、+w、+-と表記し、陰性を-表記とした。
1.材料及び方法
(1)金コロイド標識抗体の作製
2mMホウ酸緩衝液(pH9.0)で1mg/mlとなるようにPαs1CN2(FERM-BP-10264)のMAb溶液を調製した。あらかじめ0.2M炭酸カリウム溶液でpH9.0に調製した金コロイド溶液(シグマ社製)5mlにMAb溶液を500μl加え、室温で30分間反応した後、10%BSA溶液を635μlを加え、さらに15分間反応させた。遠心分離を行い、1%BSA溶液でOD525=1.0になるよう調製した。
PBSで4mg/mlとなるようにPαs1CN1(FERM-BP-10263)のMAb溶液を調製し、ニトロセルロースメンブレンに直線状に塗布し乾燥させた。その後、0.1%牛皮ゼラチンを含むTBSで37℃、1時間ブロッキング後、TBSで洗浄し乾燥させた。
抗体固定化メンブレンに加えて、被検液スポット用のガラスウール製サンプルパッド、被検液吸収用のガラスウール製吸収パッドを別途用意し、サンプルパッド、抗体固定化メンブレン、吸収パッドの順にそれぞれ貼り付け、イムノクロマトストリップとした。乳タンパクは穐山らの方法に従い、ホルスタイン種の新鮮乳より調製した。また、検出用サンプルは表5に示す配合のモデル食肉製品を供試し、加熱温度・時間、サンプルの前処理は実施例1と同様の条件で行った。
調製した金コロイド標識抗体を20μl、展開液として牛胎児血清(FBS)を30μl、測定サンプルを50μl加え、イムノクロマトストリップに供試し、検出を確認した。
次に検出結果を表6示す。判定はラインの強い方から順に+、+w、+-と表記し、陰性を-表記とした。
1.材料及び方法
1)モデル食肉製品の加熱温度・時間、サンプルの前処理、及びイムノクロマトグラフィーによる検出の確認を実施例2と同様の条件で行った他は、実施例3と同様に行った。
次に検出結果を表7及び表8に示す。判定はラインの強い方から順に+、+w、+-と表記し、陰性を-表記とした。
1.材料及び方法
(1)金コロイド標識抗体の作製
2mMホウ酸緩衝液(pH9.0)で1mg/mlとなるようにPβLG4(FERM-ABP-11238)のMAb溶液を調製した。あらかじめ0.2M炭酸カリウム溶液でpH9.0に調製した金コロイド溶液(シグマ社製)5mlにMAb溶液を500μl加え、室温で30分間反応した後、10%BSA溶液を635μlを加え、さらに15分間反応させた。遠心分離を行い、1%BSA溶液でOD525=1.0になるよう調製した。
PBSで4mg/mlとなるようにPβLG3(FERM-ABP-11237)のMAb溶液を調製し、ニトロセルロースメンブレンに直線状に塗布し乾燥させた。その後、0.1%牛皮ゼラチンを含むTBSで37℃、1時間ブロッキング後、TBSで洗浄し乾燥させた。
抗体固定化メンブレンに加えて、被検液スポット用のガラスウール製サンプルパッド、被検液吸収用のガラスウール製吸収パッドを別途用意し、サンプルパッド、抗体固定化メンブレン、吸収パッドの順にそれぞれ貼り付け、イムノクロマトストリップとした。乳タンパクは穐山らの方法に従い、ホルスタイン種の新鮮乳より調製した。また、検出用サンプルは表3に示す配合のモデル食肉製品を供試し、加熱温度・時間、サンプルの前処理は前記同様の条件で行った。
調製した金コロイド標識抗体を20μl、展開液として牛胎児血清(FBS)を30μl、測定サンプルを50μl加え、イムノクロマトストリップに供試し、検出を確認した。
次に検出結果を表9に示す。判定はラインの強い方から順に+、+w、+-と表記し、陰性を-表記とした。
1.材料及び方法
1)モデル食肉製品の加熱温度・時間、サンプルの前処理、及びイムノクロマトグラフィーによる検出の確認を実施例2と同様の条件で行った他は、実施例5と同様に行った。
次に検出結果を表10及び表11に示す。判定はラインの強い方から順に+、+w、+-と表記し、陰性を-表記とした。
1.材料及び方法
(1)金コロイド標識抗体の作製
2mMホウ酸緩衝液(pH9.0)で1mg/mlとなるようにPGL2(FERM-BP-10268)のMAb溶液を調製した。あらかじめ0.2M炭酸カリウム溶液でpH9.0に調製した金コロイド溶液(シグマ社製)5mlにMAb溶液を500μl加え、室温で30分間反応した後、10%BSA溶液を635μlを加え、さらに15分間反応させた。遠心分離を行い、1%BSA溶液でOD525=1.0になるよう調製した。
PBSで4mg/mlとなるようにPGL1(FERM-BP-10267)のMAb溶液を調製し、ニトロセルロースメンブレンに直線状に塗布し乾燥させた。その後、1.0%牛皮ゼラチンを含むTBSで37℃、1時間ブロッキング後、TBSで洗浄し乾燥させた。
抗体固定化メンブレンに加えて、被検液スポット用のガラスウール製サンプルパッド、被検液吸収用のガラスウール製吸収パッドを別途用意し、サンプルパッド、抗体固定化メンブレン、吸収パッドの順にそれぞれ貼り付け、イムノクロマトストリップとした。小麦タンパク質は、穐山らの方法に従い、市販小麦粉末より調製した。また、検出用サンプルは、表12に示す配合のモデル食肉製品を供試し、加熱温度・時間、サンプルの前処理は前記同様の条件で行った。
調製した金コロイド標識抗体を20μl、展開液として牛胎児血清(FBS)を30μl、測定サンプルを50μl加え、イムノクロマトストリップに供試し、検出を確認した。
次に検出結果を表13に示す。判定はラインの強い方から順に+、+w、+-と表記し、陰性を-表記とした。
1.材料及び方法
1)モデル食肉製品の加熱温度・時間、サンプルの前処理、及びイムノクロマトグラフィーによる検出の確認を実施例2と同様の条件で行った他は、実施例7と同様に行った。
次に検出結果を表14及び表15に示す。判定はラインの強い方から順に+、+w、+-と表記し、陰性を-表記とした。
1.材料及び方法
(1)金コロイド標識抗体の作製
2mMホウ酸緩衝液(pH9.0)で1mg/mlとなるようにPBW2(FERM-BP-10273)のMAb溶液を調製した。あらかじめ0.2M炭酸カリウム溶液でpH9.0に調製した金コロイド溶液(シグマ社製)5mlにMAb溶液を500μl加え、室温で30分間反応した後、10%BSA溶液を635μl加え、さらに15分間反応させた。遠心分離を行い、1%BSA溶液でOD525=1.0になるよう調製した。
PBSで4mg/mlとなるようにPBW5(FERM-ABP-11241)のMAb溶液を調製し、ニトロセルロースメンブレンに直線状に塗布し乾燥させた。その後、1%スキムミルクを含むTBSで37℃、1時間ブロッキング後、TBSで洗浄し乾燥させた。
抗体固定化メンブレンに加えて、被検液スポット用のガラスウール製サンプルパッド、被検液吸収用のガラスウール製吸収パッドを別途用意し、サンプルパッド、抗体固定化メンブレン、吸収パッドの順にそれぞれ貼り付け、イムノクロマトストリップとした。そばタンパク質は、穐山らの方法に従い、市販そば粉末より調製した。また、検出用サンプルは、表16に示す配合のモデル食肉製品を供試し、加熱温度・時間、サンプルの前処理は前記同様の条件で行った。
調製した金コロイド標識抗体を20μl、展開液として牛胎児血清(FBS)を30μl、測定サンプルを50μl加え、イムノクロマトストリップに供試し、検出を確認した。
次に検出結果を表17に示す。判定はラインの強い方から順に+、+w、+-と表記し、陰性を-表記とした。
1.材料及び方法
1)モデル食肉製品の加熱温度・時間、サンプルの前処理、及びイムノクロマトグラフィーによる検出の確認を実施例2と同様の条件で行った他は、実施例9と同様に行った。
次に検出結果を表18及び表19に示す。判定はラインの強い方から順に+、+w、+-と表記し、陰性を-表記とした。
1.材料及び方法
(1)金コロイド標識抗体の作製
2mMホウ酸緩衝液(pH9.0)で1mg/mlとなるようにPAh1-4(FERM-ABP-11239)のMAb溶液を調製した。あらかじめ0.2M炭酸カリウム溶液でpH9.0に調製した金コロイド溶液(シグマ社製)5mlにMAb溶液を500μl加え、室温で30分間反応した後、10%BSA溶液を635μl加え、さらに15分間反応させた。遠心分離を行い、1%BSA溶液でOD525=1.0になるよう調製した。
PBSで4mg/mlとなるようにPAh1-5(FERM-ABP-11240)のMAb溶液を調製し、ニトロセルロースメンブレンに直線状に塗布し乾燥させた。その後、1%スキムミルクを含むTBSで37℃、1時間ブロッキング後、TBSで洗浄し乾燥させた。
抗体固定化メンブレンに加えて、被検液スポット用のガラスウール製サンプルパッド、被検液吸収用のガラスウール製吸収パッドを別途用意し、サンプルパッド、抗体固定化メンブレン、吸収パッドの順にそれぞれ貼り付け、イムノクロマトストリップとした。そばタンパク質は、穐山らの方法に従い、脱脂落花生より調製した。また、検出用サンプルは、表20に示す配合のモデル食肉製品を供試し、加熱温度・時間、サンプルの前処理は前記同様の条件で行った。
調製した金コロイド標識抗体を20μl、展開液として牛胎児血清(FBS)を30μl、測定サンプルを50μl加え、イムノクロマトストリップに供試し、検出を確認した。
次に検出結果を表21に示す。判定はラインの強い方から順に+、+w、+-と表記し、陰性を-表記とした。
1.材料及び方法
1)モデル食肉製品の加熱温度・時間、サンプルの前処理、及びイムノクロマトグラフィーによる検出の確認を実施例2と同様の条件で行った他は、実施例11と同様に行った。
次に検出結果を表22及び表23に示す。判定はラインの強い方から順に+、+w、+-と表記し、陰性を-表記とした。
Claims (9)
- 変性及び未変性のアレルゲンに対するモノクローナル抗体に金コロイドを結合した金コロイド標識抗体と、前記金コロイド標識抗体と異なるエピトープを認識する変性及び未変性のアレルゲンに対するモノクローナル抗体が所定の位置に固定された展開支持体と、アレルゲンを含む食品等の被検試料から、陰イオン性界面活性剤とチオ硫酸塩、又は、陰イオン性界面活性剤と非イオン性界面活性剤を用いて抽出した変性及び未変性のアレルゲンの測定サンプルを含む展開液を用い、展開支持体に展開させた後、金コロイドの集積の有無により、アレルゲンを検出するイムノクロマト法において、ウシ胎児血清(FBS)が少なくとも10重量%含まれている展開液を用いることを特徴とするイムノクロマト法によるアレルゲンの検出方法。
- ウシ胎児血清(FBS)が少なくとも30重量%含まれている展開液を用いることを特徴とする請求項1記載のイムノクロマト法によるアレルゲンの検出方法。
- ウシ胎児血清(FBS)が少なくとも50重量%含まれている展開液を用いることを特徴とする請求項2記載のイムノクロマト法によるアレルゲンの検出方法。
- 変性及び未変性のアレルゲンに対するモノクローナル抗体が、乳アレルゲンの主要成分としてのαs1カゼイン、ホエーアレルゲンの主要成分であるβラクトグロブリン、卵白アレルゲンとしてのオボアルブミンとオボムコイド、小麦アレルゲンの主要成分としてのグリアジン、そばの主要タンパク質である分子量24kDaと76kDaのタンパク質、落花生の主要タンパク質であるArah1から選ばれる変性及び未変性のアレルゲンを特異的に認識する2種類のモノクローナル抗体であることを特徴とする請求項1~3のいずれか記載のイムノクロマト法によるアレルゲンの検出方法。
- 陰イオン性界面活性剤としてドデシル硫酸ナトリウムを用いることを特徴とする請求項1~4のいずれか記載のイムノクロマト法によるアレルゲンの検出法。
- 非イオン性界面活性剤としてTween20を用いることを特徴とする請求項1~5のいずれか記載のイムノクロマト法によるアレルゲンの検出法。
- 変性及び未変性のアレルゲンに対するモノクローナル抗体に金コロイドを結合した金コロイド標識抗体を担持させた金コロイド標識抗体担持体、前記金コロイド標識抗体と異なるエピトープを認識する変性及び未変性のアレルゲンに対するモノクローナル抗体が所定の位置に固定された展開支持体と、アレルゲンを含む食品等の被検試料から、変性及び未変性のアレルゲンを抽出するための陰イオン性界面活性剤とチオ硫酸塩、又は、陰イオン性界面活性剤と非イオン性界面活性剤を含有する緩衝液と、アレルゲンを含む食品等の被検試料から、陰イオン性界面活性剤とチオ硫酸塩、又は、陰イオン性界面活性剤と非イオン性界面活性剤を用いて抽出した変性及び未変性のアレルゲンの測定サンプルを担持させることができるサンプル用担体と、ウシ胎児血清(FBS)又はウシ胎児血清(FBS)を含む展開液とを備えたことを特徴とするイムノクロマト用アレルゲンの検出キット。
- 変性及び未変性のアレルゲンに対するモノクローナル抗体が、乳アレルゲンの主要成分としてのαs1カゼイン、ホエーアレルゲンの主要成分であるβラクトグロブリン、卵白アレルゲンとしてのオボアルブミンとオボムコイド、小麦アレルゲンの主要成分としてのグリアジン、そばの主要タンパク質である分子量24kDaと76kDaのタンパク質、落花生の主要タンパク質であるArah1から選ばれる変性及び未変性のアレルゲンを特異的に認識する2種類のモノクローナル抗体であることを特徴とする請求項7記載のイムノクロマト用アレルゲンの検出キット。
- 陰イオン性界面活性剤としてドデシル硫酸ナトリウムを含有することを特徴とする請求項7又は8記載のイムノクロマト用アレルゲンの検出キット。
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CN103439502A (zh) * | 2013-06-28 | 2013-12-11 | 北京新华联协和药业有限责任公司 | 一种快速检测特异性抗体IgE试剂盒及其制备方法 |
JP2016211967A (ja) * | 2015-05-08 | 2016-12-15 | プリマハム株式会社 | イムノクロマト法によるアレルゲンの検出方法 |
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Citations (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH04262796A (ja) * | 1991-02-18 | 1992-09-18 | Green Cross Corp:The | 基質液の安定化法及び基質液 |
JPH0510950A (ja) | 1990-08-31 | 1993-01-19 | Japan Synthetic Rubber Co Ltd | イムノクロマトグラフ法 |
JPH1042869A (ja) * | 1996-08-01 | 1998-02-17 | Eiken Chem Co Ltd | ビリルビンオキシダーゼの安定化方法 |
WO2002037099A1 (fr) * | 2000-10-27 | 2002-05-10 | International Reagents Corporation | Procede de diagnostic de la nephropathie |
JP2003344406A (ja) | 2002-05-24 | 2003-12-03 | Towns:Kk | イムノクロマトグラフィー用展開溶媒、測定法およびキット |
JP2005106811A (ja) * | 2004-08-13 | 2005-04-21 | Morinaga & Co Ltd | イムノアッセイ |
JP2006071509A (ja) * | 2004-09-02 | 2006-03-16 | Nippon Meat Packers Inc | 食品成分抽出液 |
JP2006317226A (ja) * | 2005-05-11 | 2006-11-24 | Morinaga & Co Ltd | 加熱処理された動物性組織由来原料の検出試薬および検出方法 |
JP2007278773A (ja) | 2006-04-04 | 2007-10-25 | Prima Meat Packers Ltd | イムノクロマト法によるアレルゲンの検出方法 |
WO2009069779A1 (ja) * | 2007-11-30 | 2009-06-04 | Morinaga & Co., Ltd. | 食品成分抽出方法及び食品検査方法並びに食品検査キット |
Family Cites Families (13)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1130567C (zh) * | 1997-10-17 | 2003-12-10 | 库特国际公司 | 测量血红蛋白和区分白细胞的不含氰化物的试剂及方法 |
JP2000065820A (ja) * | 1998-08-19 | 2000-03-03 | Allergen Free Technology Kenkyusho:Kk | タンパク質抽出用水溶液及び抽出方法並びにアレルゲン分析方法 |
JP2001305134A (ja) * | 2000-04-21 | 2001-10-31 | Shinto Fine Co Ltd | ダニアレルゲンの簡易検査キット及びそれを用いる検知方法 |
JP2002267670A (ja) * | 2001-03-07 | 2002-09-18 | Asahi Beer Yakuhin Kk | 特異的結合を利用する検体分析方法 |
ES2292795T3 (es) * | 2001-07-17 | 2008-03-16 | Keratec Limited | Produccion de derivados de queratina solubles. |
US8361460B2 (en) * | 2001-09-05 | 2013-01-29 | Nippon Meat Packers, Inc. | Food allergens, method of detecting food allergens and method of detecting food allergy-inducing foods |
US20060228769A1 (en) * | 2002-08-13 | 2006-10-12 | Inc., Admn. Agncy, Nat. Agr. & Bio-Orntd Rsrch Org | Method of detecting allergen protein |
JP3920741B2 (ja) * | 2002-08-28 | 2007-05-30 | 森永乳業株式会社 | 物質の検出試薬及び検出方法 |
JP3600231B1 (ja) * | 2003-09-30 | 2004-12-15 | 森永製菓株式会社 | イムノアッセイ |
WO2005085847A1 (ja) * | 2004-03-05 | 2005-09-15 | Prima Meat Packers, Ltd. | アレルゲンの検出方法 |
JP2005291783A (ja) | 2004-03-31 | 2005-10-20 | Denka Seiken Co Ltd | 免疫測定に供する検体浮遊液調製用媒体組成物及びそれを用いる免疫測定方法 |
CN1314952C (zh) * | 2004-11-23 | 2007-05-09 | 中国检验检疫科学研究院 | 用于固相膜免疫分析方法流动相的样本处理制剂 |
CN101000349B (zh) * | 2006-12-31 | 2011-08-24 | 浙江伊利康生物技术有限公司 | 前白蛋白检测试剂盒 |
-
2010
- 2010-02-23 AU AU2010216932A patent/AU2010216932B2/en not_active Ceased
- 2010-02-23 KR KR1020117018533A patent/KR101331817B1/ko not_active IP Right Cessation
- 2010-02-23 US US13/202,016 patent/US20110306150A1/en not_active Abandoned
- 2010-02-23 EP EP10743598.4A patent/EP2400301B1/en not_active Not-in-force
- 2010-02-23 CN CN201610040264.9A patent/CN105651994B/zh active Active
- 2010-02-23 JP JP2011500539A patent/JP5735411B2/ja active Active
- 2010-02-23 WO PCT/JP2010/001209 patent/WO2010095469A1/ja active Application Filing
- 2010-02-23 CN CN201080007948.XA patent/CN102317780B/zh active Active
- 2010-02-23 CA CA2752126A patent/CA2752126C/en not_active Expired - Fee Related
- 2010-02-23 NZ NZ594549A patent/NZ594549A/xx not_active IP Right Cessation
Patent Citations (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH0510950A (ja) | 1990-08-31 | 1993-01-19 | Japan Synthetic Rubber Co Ltd | イムノクロマトグラフ法 |
JPH04262796A (ja) * | 1991-02-18 | 1992-09-18 | Green Cross Corp:The | 基質液の安定化法及び基質液 |
JPH1042869A (ja) * | 1996-08-01 | 1998-02-17 | Eiken Chem Co Ltd | ビリルビンオキシダーゼの安定化方法 |
WO2002037099A1 (fr) * | 2000-10-27 | 2002-05-10 | International Reagents Corporation | Procede de diagnostic de la nephropathie |
JP2003344406A (ja) | 2002-05-24 | 2003-12-03 | Towns:Kk | イムノクロマトグラフィー用展開溶媒、測定法およびキット |
JP2005106811A (ja) * | 2004-08-13 | 2005-04-21 | Morinaga & Co Ltd | イムノアッセイ |
JP2006071509A (ja) * | 2004-09-02 | 2006-03-16 | Nippon Meat Packers Inc | 食品成分抽出液 |
JP2006317226A (ja) * | 2005-05-11 | 2006-11-24 | Morinaga & Co Ltd | 加熱処理された動物性組織由来原料の検出試薬および検出方法 |
JP2007278773A (ja) | 2006-04-04 | 2007-10-25 | Prima Meat Packers Ltd | イムノクロマト法によるアレルゲンの検出方法 |
WO2009069779A1 (ja) * | 2007-11-30 | 2009-06-04 | Morinaga & Co., Ltd. | 食品成分抽出方法及び食品検査方法並びに食品検査キット |
Non-Patent Citations (2)
Title |
---|
AKIYAMA ET AL.: "Inter-laboratory Evaluation Studies of Notified ELISA Methods for Allergic Substances (Egg", J. FOOD HYG. SOC. JAPAN, vol. 44, 2003, pages 213 - 219 |
See also references of EP2400301A4 |
Cited By (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103439502A (zh) * | 2013-06-28 | 2013-12-11 | 北京新华联协和药业有限责任公司 | 一种快速检测特异性抗体IgE试剂盒及其制备方法 |
JP2016211967A (ja) * | 2015-05-08 | 2016-12-15 | プリマハム株式会社 | イムノクロマト法によるアレルゲンの検出方法 |
US10753928B2 (en) | 2015-12-14 | 2020-08-25 | Morinaga Institute Of Biological Science, Inc. | Protein detection method, and protein immunoassay method |
JP2018072209A (ja) * | 2016-10-31 | 2018-05-10 | プリマハム株式会社 | チョコレート試料中の小麦アレルゲンの検出方法 |
JP2018077171A (ja) * | 2016-11-10 | 2018-05-17 | プリマハム株式会社 | チョコレート試料中のアレルゲンの検出方法 |
JP2018193372A (ja) * | 2018-05-29 | 2018-12-06 | 株式会社森永生科学研究所 | 抗ペプチド抗体の製造方法及び設計方法 |
JP2021032760A (ja) * | 2019-08-27 | 2021-03-01 | 日鉄ケミカル&マテリアル株式会社 | プロゾーン現象の抑制方法 |
JP7458583B2 (ja) | 2019-08-27 | 2024-04-01 | 日鉄ケミカル&マテリアル株式会社 | プロゾーン現象の抑制方法 |
WO2022102607A1 (ja) * | 2020-11-10 | 2022-05-19 | プリマハム株式会社 | アレルゲン検出キット |
KR20230104883A (ko) | 2020-11-10 | 2023-07-11 | 뿌리마하무가부시끼가이샤 | 알레르겐 검출 키트 |
Also Published As
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AU2010216932B2 (en) | 2012-12-06 |
EP2400301B1 (en) | 2013-06-05 |
EP2400301A1 (en) | 2011-12-28 |
KR101331817B1 (ko) | 2013-11-22 |
CN102317780B (zh) | 2016-02-17 |
CN105651994A (zh) | 2016-06-08 |
NZ594549A (en) | 2013-02-22 |
CA2752126C (en) | 2014-04-29 |
CA2752126A1 (en) | 2010-08-26 |
JP5735411B2 (ja) | 2015-06-17 |
AU2010216932A1 (en) | 2011-09-01 |
KR20110114626A (ko) | 2011-10-19 |
CN105651994B (zh) | 2019-03-08 |
EP2400301A4 (en) | 2012-06-20 |
JPWO2010095469A1 (ja) | 2012-08-23 |
US20110306150A1 (en) | 2011-12-15 |
CN102317780A (zh) | 2012-01-11 |
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