WO2010113943A1 - 免疫分析方法及びそのための試薬 - Google Patents
免疫分析方法及びそのための試薬 Download PDFInfo
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- WO2010113943A1 WO2010113943A1 PCT/JP2010/055693 JP2010055693W WO2010113943A1 WO 2010113943 A1 WO2010113943 A1 WO 2010113943A1 JP 2010055693 W JP2010055693 W JP 2010055693W WO 2010113943 A1 WO2010113943 A1 WO 2010113943A1
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- immunoassay
- fatty acid
- acid ester
- sorbitan fatty
- polyoxyethylene sorbitan
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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
- G01N33/531—Production of immunochemical test materials
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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
- G01N33/5306—Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding
-
- 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
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
Definitions
- the present invention relates to a novel immunoassay method useful for measurement of a low-concentration sample and a reagent therefor.
- Immunological analysis methods are widely used because they can be easily and quickly measured by using an automatic analyzer in clinical tests of serum, plasma, urine and the like.
- the stability of the reagent will decrease, and self-aggregation will cause the agglutination reaction to occur during storage of the reagent, or nonspecific agglutination will occur and even blank samples In some cases, the agglutination reaction proceeded.
- the upper limit is set for the amount of change in absorbance, so the sensitivity in the high value region is limited and a wide measurement region can be obtained. There may not be.
- an object of the present invention is to provide a novel method capable of expanding the measurement area in immunoassay.
- the inventor of the present application has made it possible to suppress variations in the measured values of the blank by allowing polyoxyethylene alkyl ether sulfate and polyoxyethylene sorbitan fatty acid ester to coexist in the reaction system during immunoassay.
- the measured value also decreased, and it was found that the measurement accuracy in the low concentration range of the test substance could be increased, and the present invention was completed.
- the present invention provides an immunoassay method including coexisting polyoxyethylene alkyl ether sulfate and polyoxyethylene sorbitan fatty acid ester in the reaction system.
- the present invention also provides a method for measuring a marker protein in a sample by the immunoassay method of the present invention.
- the present invention provides a method for stabilizing the measurement value by immunoassay, which comprises coexisting polyoxyethylene alkyl ether sulfate and polyoxyethylene sorbitan fatty acid ester in the reaction system in the immunoassay method.
- the present invention provides an immunoassay reagent containing polyoxyethylene alkyl ether sulfate and polyoxyethylene sorbitan fatty acid ester.
- the present invention provides an immunoassay kit containing the immunoassay reagent of the present invention. Furthermore, the present invention provides a measurement value stabilizer for immunoassay comprising polyoxyethylene alkyl ether sulfate and polyoxyethylene sorbitan fatty acid ester.
- a novel means capable of increasing the measurement accuracy in a low concentration range of a test substance in immunoassay.
- the method of the present invention only by adding a predetermined substance to the reaction system, the variation (standard deviation) of the measured value of the blank is suppressed, and the measured value is stabilized.
- the present invention is advantageous for micro-quantification of a substance present in a trace amount in a specimen.
- the present invention is characterized in that immunoassay is performed in the presence of polyoxyethylene alkyl ether sulfate and polyoxyethylene sorbitan fatty acid ester.
- % in this specification is a mass reference
- measurement includes detection, quantification, and semi-quantification.
- the polyoxyethylene alkyl ether sulfate is not particularly limited as long as it is usually used as a surfactant.
- the alkyl moiety may be linear or branched and may contain one or more unsaturated bonds. It may be an inert alkyl group in which one or more carbons are replaced with oxygen, such as an alkoxy group. A part of the alkyl group may be substituted with an aryl group (such as a phenyl group) or a halogen atom.
- the number of carbon atoms in the alkyl moiety is not particularly limited, but is usually 1-30, preferably 5-20, more preferably 10-15.
- a particularly preferable alkyl moiety is a linear carbon chain containing a saturated or several unsaturated bond, and specific examples thereof include a lauryl group.
- the sulfate portion may be a monovalent or higher valent salt, and specific examples include alkali metal salts such as sodium salt and potassium salt, alkaline earth metal salts such as calcium salt and magnesium salt, ammonium salt and amine salt.
- alkali metal salts such as sodium salt and potassium salt
- alkaline earth metal salts such as calcium salt and magnesium salt
- ammonium salt and amine salt A triethanolamine salt and a sodium salt are preferably used.
- the number of oxyethylene units in the molecule of polyoxyethylene alkyl ether sulfate is not particularly limited, but is usually about 1 to 10.
- the polyoxyethylene alkyl ether sulfate may have a single composition or a mixture of two or more.
- a commercially available product sold as a surfactant can be preferably used as the polyoxyethylene alkyl ether sulfate.
- the polyoxyethylene sorbitan fatty acid ester is not particularly limited as long as it is usually used as a surfactant.
- the alkyl portion of the fatty acid may be linear or branched and may contain one or more unsaturated bonds. It may be an inert alkyl group in which one or more carbons are replaced with oxygen, such as an alkoxy group. A part of the alkyl group may be substituted with a halogen atom or the like.
- the number of carbon atoms in the alkyl moiety is not particularly limited, but is usually 1-30, preferably 5-25, more preferably 10-20.
- a particularly preferred alkyl moiety is a linear or branched carbon chain containing a saturated or several unsaturated bonds, and specific examples of the fatty acid moiety include oleic acid, stearic acid, lauric acid, palmitic acid and the like. Can be mentioned.
- the number of oxyethylene units in the molecule of the polyoxyethylene sorbitan fatty acid ester is not particularly limited, but is usually about 1 to 100, preferably about 5 to 60.
- the composition of the polyoxyethylene sorbitan fatty acid ester may be a single composition or a mixture of two or more. Specific examples of the composition include, but are not limited to, polyoxyethylene sorbitan monooleate and polyoxyethylene sorbitan monostearate.
- polyoxyethylene sorbitan fatty acid ester a commercially available product sold as a surfactant can be preferably used.
- a polyoxyethylene sorbitan fatty acid ester having an HLB value of 10 to 20 and a number average molecular weight of 1000 to 50000 has a remarkable effect of addition and is preferably used.
- polyoxyethylene alkyl ether sulfate and polyoxyethylene sorbitan fatty acid ester are added into the reaction system at any stage from the start of the antigen-antibody reaction to the end of detection / quantification of the antigen-antibody reaction amount.
- the polyoxyethylene alkyl ether sulfate and the polyoxyethylene sorbitan fatty acid ester can be preferably added to the reaction system before or simultaneously with the start of the antigen-antibody reaction. For example, it may be added when the specimen is diluted, or may be added when the antibody or antigen is mixed with the specimen.
- polyoxyethylene alkyl ether sulfate and polyoxyethylene sorbitan fatty acid ester contain previously in the various reagents used for an immunoassay.
- these substances can be preliminarily contained in a buffer solution for diluting a specimen, a reagent containing an antibody or an antigen, or the like.
- a polyoxyethylene alkyl ether sulfate and a polyoxyethylene sorbitan fatty acid ester are contained in a reagent containing insoluble carrier particles (sensitized particles) to which antibodies or antigens are immobilized (sensitized). I can leave.
- polyoxyethylene alkyl ether sulfate and polyoxyethylene sorbitan fatty acid ester coexisting at the time of the antigen-antibody reaction are lost. It is preferable to appropriately add polyoxyethylene alkyl ether sulfate and polyoxyethylene sorbitan fatty acid ester to the reaction system, such as using a buffer containing polyoxyethylene sorbitan fatty acid ester.
- the concentration of polyoxyethylene alkyl ether sulfate in the reaction system is preferably 0.0005 to 0.25%, more preferably 0.0125 to 0.25%.
- the concentration of the polyoxyethylene sorbitan fatty acid ester in the reaction system is preferably 0.0005 to 0.25%, more preferably 0.0005 to 0.005%.
- it can be contained in the immunoassay reagent so that the concentration in the reaction system becomes the above-mentioned concentration.
- the concentration of polyoxyethylene alkyl ether sulfate in the reagent is preferably 0.001 to 0.5%, more preferably 0.025 to 0.5%.
- the concentration of polyoxyethylene sorbitan fatty acid ester in the reagent is preferably 0.001 to 0.5%, more preferably 0.001 to 0.01%.
- the immunoassay technique itself is well known.
- any known immunoassay method may be used.
- immunoaggregation is preferable, and latex aggregation using latex particles as insoluble carrier particles is particularly preferable.
- Methods for detecting the aggregation of sensitized particles in the immunoaggregation method are well known, and also in the present invention, known methods such as a method for detecting absorbance or light scattering due to aggregation of sensitized particles can be used.
- immunoturbidimetry TIA method, latex agglutination method
- colorimetric method RPLA method
- CL method immunochromatography method
- the turbidimetric method and the colorimetric method with high sensitivity and good quantitative accuracy are preferably used. It is done.
- the insoluble carrier particles used are not particularly limited, and may be well-known ones conventionally used for immunoassay reagents.
- latex particles such as polyethylene and polystyrene, particles such as alumina particles, silica particles, gold colloid, and magnetic particles can be used.
- latex particles, particularly polystyrene latex particles are preferably used.
- the size of the latex particles is not particularly limited, but the particle size is preferably 30 to 600 nm.
- C-reactive protein C-reactive protein
- FER ferritin
- Mb myoglobin
- PSA prostate specific antigen
- BMG ⁇ -2 microglobulin
- marker proteins such as megalin and podocalyxin
- bacteria such as Escherichia coli
- viruses such as influenza virus, norovirus, and RS virus
- antibodies to these C-reactive protein
- a preferable measurement object is a marker protein that serves as a marker for various diseases.
- the specimen used for the immunoassay is not particularly limited as long as it can contain the measurement object, but it is a body fluid such as blood, serum, plasma, urine, feces, saliva, tissue fluid, spinal fluid, and swab fluid, or a diluted product thereof. Furthermore, blood, serum, plasma, urine, stool, cerebrospinal fluid or a dilution thereof is preferable.
- the blank sample used in the immunoassay is not particularly limited as long as it does not contain the measurement target, but purified water, physiological saline, buffer, negative sample or a diluted sample thereof is preferable.
- the immunoagglutination method itself is well known and need not be explained here, but briefly explained as an example of measuring an antigen in a test sample.
- the reacting antibody or antigen-binding fragment thereof is immobilized.
- the immobilization method is also known, and is performed by a known method such as physical adsorption or covalent bond.
- a plurality of standard samples containing antigens to be measured at various known concentrations are prepared, and the change or rate of change in absorbance is measured by using the above-described method.
- a calibration curve is drawn by plotting the concentration of the antigen to be measured in the standard sample on the horizontal axis and the change or rate of change in the measured absorbance on the vertical axis.
- the various reagents for immunoassay previously containing polyoxyethylene alkyl ether sulfate and polyoxyethylene sorbitan fatty acid ester can be provided as immunoassay reagents used in the method of the present invention.
- the immunoassay reagent of the present invention includes various reagents used for immunoassay. For example, specimen dilution liquid, antibody / antigen dilution liquid, immobilized antibody / antigen, sensitized particle suspension, washing liquid, enzyme liquid, substrate liquid, test substance standard solution for preparing a calibration curve, and the like can be mentioned.
- an immunoagglutination reagent such as a latex agglutination reagent is preferred, and specifically, a reagent containing sensitized particles is preferred.
- concentration of the sensitizing particles in the immunoassay reagent is not particularly limited, but is preferably 0.01 to 0.5%.
- the amount of antibody and the amount of antigen in the sensitized particle suspension may be as usual, and are not particularly limited. For example, in the case of antibody-sensitized latex, the amount of antibody is 0.01 to 2.0 mg / mL in the latex suspension. It is preferable that
- the concentration of polyoxyethylene alkyl ether sulfate in the immunoassay reagent of the present invention is preferably 0.0005 to 0.25%, more preferably 0.0125 to 0.25%.
- the concentration of the polyoxyethylene sorbitan fatty acid ester in the immunoassay reagent is preferably 0.0005 to 0.25%, more preferably 0.0005 to 0.005%.
- polyoxyethylene alkyl ether sulfates and polyoxyethylene sorbitan fatty acid esters can be provided as measurement value stabilizers for immunoassay.
- the stabilization of the measured value here means that the standard deviation of the measured value is reduced as described above.
- the stabilizer is preferably used by being added to various reagents used for immunoassay.
- the above-described immunoassay reagent of the present invention can be provided as an immunoassay kit together with ordinary reagents and the like not containing polyoxyethylene alkyl ether sulfate and polyoxyethylene sorbitan fatty acid ester.
- the immunoassay kit of the present invention may be a kit in which normal immunoassay reagents and the above-described measurement value stabilizer of the present invention are combined.
- Sensitizing particles obtained by supporting 0.08 mg of any one of anti-CRP antibody, anti-FER antibody and anti-Mb antibody with respect to 1 mL of polystyrene latex suspension having an average particle diameter of 190 nm are added to buffer solution (Tris, pH 8.0). The suspension was suspended to 1% to prepare a latex suspension.
- Surfactants were added to the latex suspension as described below to prepare measuring reagents for each protein (CRP, FER, Mb). As a comparative example, a latex suspension without adding the surfactant component was used.
- the automatic analyzer used Hitachi 7180 type automatic analyzer for automatic measurement by the end point method.
- the measurement of the physiological saline (blank) sample solution was performed 10 times using the above-described measurement reagent. 120 ⁇ L of a buffer solution (Tris, pH 8.5) was added to 2.4 ⁇ L of the sample solution, and this mixed solution was stirred and mixed at 37 ° C. After leaving for 5 minutes, 120 ⁇ L of the measurement reagent prepared above was added, and further stirred and mixed at 37 ° C. The aggregation reaction for about 5 minutes was measured as the amount of change in absorbance, and the standard deviation was calculated.
- a buffer solution Tris, pH 8.5
- FIG. 1 shows the change in absorbance over time when the absorbance immediately after the addition of the measurement reagent is 0.000 Abs.
- the reason why the accuracy of the blank is improved is that, by adding polyoxyethylene alkyl ether triethanolamine sulfate and polyoxyethylene sorbitan fatty acid ester, there is a variation in absorbance due to the elapsed time after adding the measurement reagent. Suppressed. The effect of suppressing this variation does not change even when measured twice, indicating that there is reproducibility.
- a surfactant was added to the latex suspension as described below to prepare three types of CRP measurement reagents.
- Reagent No. 2-1 (Example) Latex suspension + Surfactant 1 0.1% Surfactant 2 0.005% Reagent No. 2-2 (Comparative example) Latex suspension + Surfactant 1 0.1% Reagent No. 2-3 (comparative example) Latex suspension only surfactant 1: Polyoxyethylene alkyl ether sulfate triethanolamine (supra)
- Surfactant 2 Polyoxyethylene sorbitan fatty acid ester (supra)
- the automatic analyzer used Hitachi 7180 type automatic analyzer for automatic measurement by the end point method.
- the sample solution was measured 10 times each using the above 3 CRP measuring reagents.
- physiological saline (blank) and physiological saline (CRP) containing CRP at a concentration of 0.005 mg / dL were used as a sample solution.
- 120 ⁇ L of a buffer solution (Tris, pH 8.5) was added to 2.4 ⁇ L of the sample solution, and this mixed solution was stirred and mixed at 37 ° C. After leaving for 5 minutes, 120 ⁇ L of the CRP measurement reagent was added, and the mixture was further stirred and mixed at 37 ° C. The aggregation reaction for about 5 minutes was measured as the amount of change in absorbance.
- the standard deviation SD of the measurement data of the blank and the CRP sample is reduced, and the measured values of the CRP concentrations 0 mg / dL (blank) and 0.005 mg / dL (CRP sample) calculated by the above formula are reduced.
- the difference has grown.
- the standard deviation SD value was particularly small on the blank side. As described above, since the difference between the measured values of the CRP concentration of 0 mg / dL and the CRP concentration of 0.005 mg / dL in the specimen was enlarged, it was shown that the measurable region on the low concentration side was expanded.
- Surfactants were added to the latex suspension as described below to prepare 5 types of CRP measurement reagents with varying concentrations of surfactant 1.
- Reagent No. 3-1 (Comparative example) Latex suspension + surfactant 10% Surfactant 2 0.005% Reagent No. 3-2 (Example) Latex suspension + surfactant 1 0.01% Surfactant 2 0.005% Reagent No. 3-3 (Example) Latex suspension + surfactant 1 0.05% Surfactant 2 0.005% Reagent No. 3-4 (Example) Latex suspension + Surfactant 1 0.10% Surfactant 2 0.005% Reagent No.
- Surfactant 1 Polyoxyethylene alkyl ether sulfate triethanolamine (supra)
- Surfactant 2 Polyoxyethylene sorbitan fatty acid ester (supra)
- the automatic analyzer used Hitachi 7180 type automatic analyzer for automatic measurement by the end point method.
- the measurement of the physiological saline (blank) sample solution was performed 5 times for each reagent using the above five CRP measurement reagents, and the average value was obtained.
- 120 ⁇ L of a buffer solution (Tris, pH 8.5) was added to 2.4 ⁇ L of the sample solution, and this mixed solution was stirred and mixed at 37 ° C.
- 120 ⁇ L of the CRP measurement reagent was added, and the mixture was further stirred and mixed at 37 ° C.
- the aggregation reaction for about 5 minutes was measured as the amount of change in absorbance, and the average value of the amount of change in absorbance was compared.
Abstract
Description
C-反応性タンパク質(CRP)、フェリチン(FER)及びミオグロビン(Mb)に対する抗体を用いて、以下の通りに免疫凝集法による測定試薬を調製した。
測定試薬(実施例)
ラテックス浮遊液
+ 界面活性剤1 0.1%
界面活性剤2 0.005%
測定試薬(比較例)
ラテックス浮遊液のみ
界面活性剤1:
ポリオキシエチレンアルキルエーテル硫酸トリエタノールアミン(花王製「エマール20T」(商品名)、ポリオキシエチレンラウリルエ一テル硫酸トリエタノールアミンを主体)
界面活性剤2:
ポリオキシエチレンソルビタン脂肪酸エステル(和光純薬製「Tween80」(商品名)、ポリオキシエチレンソルビタンモノオレエートを主体、HLB=15、数平均分子量5000)
自動分析装置は日立7180型自動分析装置によりエンドポイント法で自動測定を行った。
測定結果を下記表1に示す。また、測定試薬添加直後の吸光度を0.000Absとした場合の経過時間における吸光度の変化を図1に示す。
ポリオキシエチレンアルキルエーテル硫酸トリエタノールアミン及びポリオキシエチレンソルビタン脂肪酸エステルを添加することでブランクの標準偏差が小さくなり、ブランクの精度が向上することが示された。また、上記成分の添加効果は、抗体の種類に関わらず得られることが示された。
抗CRP抗体を平均粒径150nmのポリスチレンラテックス浮遊液1mLに対し0.06mg担持させてなる感作粒子を、緩衝液(トリス、pH8.0)に0.12%となるように懸濁し、ラテックス浮遊液を調製した。
試薬No.2-1(実施例)
ラテックス浮遊液
+ 界面活性剤1 0.1%
界面活性剤2 0.005%
試薬No.2-2(比較例)
ラテックス浮遊液
+ 界面活性剤1 0.1%
試薬No.2-3(比較例)
ラテックス浮遊液のみ
界面活性剤1:
ポリオキシエチレンアルキルエーテル硫酸トリエタノールアミン(前掲)
界面活性剤2:
ポリオキシエチレンソルビタン脂肪酸エステル(前掲)
自動分析装置は日立7180型自動分析装置によりエンドポイント法で自動測定を行った。
各サンプルの10回測定のデータより平均値と標準偏差を算出し、下記式により数値(以下、単に「差」という)を求めて測定精度を評価した。下記式により求められる差は、ブランクとCRP試料との間の測定値の有意差の指標となるものであり、差の値が大きいほど測定値間の有意差が大きいことを示す。
[CRPの平均値-2×(CRPのSD)]-[ブランクの平均値+2×(ブランクのSD)]
界面活性剤を添加することでブランク及びCRP試料の測定データの標準偏差SDが小さくなり、上記式で算出したCRP濃度0mg/dL(ブランク)と0.005mg/dL(CRP試料)の測定値の差が大きくなった。標準偏差SD値は、ブランク側で特に小さくなった。このように、検体中のCRP濃度0mg/dLとCRP濃度0.005mg/dLの測定値の差が拡大したので、低濃度側の測定可能領域が広がることが示された。
抗CRP抗体を平均粒径190nmのポリスチレンラテックス浮遊液1mLに対し0.04mg担持させてなる感作粒子を、緩衝液(トリス、pH8.0)に0.1%となるように懸濁し、ラテックス浮遊液を調製した。
試薬No.3-1(比較例)
ラテックス浮遊液
+ 界面活性剤1 0%
界面活性剤2 0.005%
試薬No.3-2(実施例)
ラテックス浮遊液
+ 界面活性剤1 0.01%
界面活性剤2 0.005%
試薬No.3-3(実施例)
ラテックス浮遊液
+ 界面活性剤1 0.05%
界面活性剤2 0.005%
試薬No.3-4(実施例)
ラテックス浮遊液
+ 界面活性剤1 0.10%
界面活性剤2 0.005%
試薬No.3-5(実施例)
ラテックス浮遊液
+ 界面活性剤1 0.20%
界面活性剤2 0.005%
界面活性剤1:
ポリオキシエチレンアルキルエーテル硫酸トリエタノールアミン(前掲)
界面活性剤2:
ポリオキシエチレンソルビタン脂肪酸エステル(前掲)
自動分析装置は日立7180型自動分析装置によりエンドポイント法で自動測定を行った。
ポリオキシエチレンアルキルエーテル硫酸トリエタノールアミンを添加することで生理食塩水(ブランク)測定時の吸光度変化量が小さくなり、非特異的凝集が著しく抑制されていたことが確認できた。これにより、測定対象濃度が低い領域での試薬の安定性が向上することが示された。
Claims (17)
- ポリオキシエチレンアルキルエーテル硫酸塩及びポリオキシエチレンソルビタン脂肪酸エステルを反応系内に共存させることを含む免疫分析方法。
- 前記ポリオキシエチレンアルキルエーテル硫酸塩の濃度が0.0005~0.25%である請求項1記載の免疫分析方法。
- 前記ポリオキシエチレンソルビタン脂肪酸エステルの濃度が0.0005~0.005%である請求項1又は2記載の免疫分析方法。
- 前記ポリオキシエチレンソルビタン脂肪酸エステルのHLB値が10~20である請求項1ないし3のいずれか1項に記載の免疫分析方法。
- 前記ポリオキシエチレンソルビタン脂肪酸エステルの数平均分子量が1000~50000である請求項1ないし4のいずれか1項に記載の免疫分析方法。
- 前記ポリオキシエチレンアルキルエーテル硫酸塩がポリオキシエチレンラウリルエ一テル硫酸トリエタノールアミンである請求項1ないし5のいずれか1項に記載の免疫分析方法。
- 前記ポリオキシエチレンソルビタン脂肪酸エステルがポリオキシエチレンソルビタンモノオレエートである請求項1ないし6のいずれか1項に記載の免疫分析方法。
- 免疫凝集法である請求項1ないし7のいずれか1項に記載の免疫分析方法。
- ラテックス凝集法である請求項9記載の免疫分析方法。
- 請求項1ないし9のいずれか1項に記載の方法による検体中のマーカータンパク質測定方法。
- 免疫分析方法において、ポリオキシエチレンアルキルエーテル硫酸塩及びポリオキシエチレンソルビタン脂肪酸エステルを反応系内に共存させることを含む、免疫分析による測定値を安定化する方法。
- ポリオキシエチレンアルキルエーテル硫酸塩及びポリオキシエチレンソルビタン脂肪酸エステルを含有する免疫分析試薬。
- 前記ポリオキシエチレンアルキルエーテル硫酸塩の濃度が0.0005~0.25%である請求項12記載の免疫分析試薬。
- 前記ポリオキシエチレンソルビタン脂肪酸エステルの濃度が0.0005~0.005%である請求項12又は13記載の免疫分析試薬。
- 請求項12ないし14のいずれか1項に記載の免疫分析試薬を含む免疫分析キット。
- ポリオキシエチレンアルキルエーテル硫酸塩及びポリオキシエチレンソルビタン脂肪酸エステルからなる免疫分析用測定値安定化剤。
- 免疫分析試薬に添加された形態にある請求項16記載の測定値安定化剤。
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CN201080014485.XA CN102369441B (zh) | 2009-03-31 | 2010-03-30 | 免疫分析方法及用于该方法的试剂 |
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US10627393B2 (en) | 2012-07-31 | 2020-04-21 | Sekisui Medical Co., Ltd. | Latex agglutination inhibition immunoassay |
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