WO1995013539A1 - Methode de dosage d'anticorps anti-interferon et reactif utilise dans ce dernier - Google Patents

Methode de dosage d'anticorps anti-interferon et reactif utilise dans ce dernier Download PDF

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Publication number
WO1995013539A1
WO1995013539A1 PCT/JP1993/001657 JP9301657W WO9513539A1 WO 1995013539 A1 WO1995013539 A1 WO 1995013539A1 JP 9301657 W JP9301657 W JP 9301657W WO 9513539 A1 WO9513539 A1 WO 9513539A1
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WO
WIPO (PCT)
Prior art keywords
interferon
antibody
solid
antigen
antibody complex
Prior art date
Application number
PCT/JP1993/001657
Other languages
English (en)
Japanese (ja)
Inventor
Yasushi Nakamura
Koji Ushizawa
Original Assignee
Daiichi Pure Chemicals 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
Priority to JP4261627A priority Critical patent/JPH06109728A/ja
Priority claimed from JP4261627A external-priority patent/JPH06109728A/ja
Application filed by Daiichi Pure Chemicals Co., Ltd. filed Critical Daiichi Pure Chemicals Co., Ltd.
Priority to PCT/JP1993/001657 priority patent/WO1995013539A1/fr
Publication of WO1995013539A1 publication Critical patent/WO1995013539A1/fr

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6863—Cytokines, i.e. immune system proteins modifying a biological response such as cell growth proliferation or differentiation, e.g. TNF, CNF, GM-CSF, lymphotoxin, MIF or their receptors
    • G01N33/6866—Interferon

Definitions

  • the present invention relates to a method for simply and accurately quantifying an anti-infatin antibody which is useful as a factor for determining a clinical effect upon treatment with interferon by immunological means.
  • This anti-interferon antibody quantification method is based on
  • the measurement method by ELISA is excellent in simplicity
  • the conventional ELISA method involves directly immobilizing the interferon, which is an antigen, on a solid phase carrier. It is hard to say that all antigen-antibody complexes that need to be purified and that may be present in the sample can be measured simultaneously.
  • an object of the present invention is to provide a method for quantifying an anti-interfuron antibody with high accuracy by a simple operation.
  • the present inventors had studied various methods for quantifying anti-interferin ⁇ antibody by immunological means using immobilized antigen.
  • the anti-interferin antibody was previously bound to a solid-phase carrier. If the immobilized antigen-antibody complex obtained by binding the antigen, which is an antigen, is used as the immobilized antigen, it is not necessary to purify the antigen to be immobilized.
  • the present inventors have found that not only a single antibody but also an interferon-anti-interfuron antibody complex present in a sample can be simultaneously and accurately quantified, thereby completing the present invention.
  • the present invention relates to a method for quantifying an anti-interferon antibody using an immunoreaction between a solid-phased antigen and a test solution, wherein the solid-phased carrier comprises an anti-interferon antibody, and then an interferon.
  • a solid-phased antigen-antibody complex obtained by successively binding anti-interferon antibodies and an interferon-anti-interferon antibody complex in a test solution, characterized in that It is.
  • the present invention provides an anti-interferon antibody and an interferon-anti-interferon antibody containing a surface-phased antigen-antibody complex obtained by sequentially binding an anti-interferon antibody to a solid phase carrier and then interferon. It relates to a reagent for quantifying the complex.
  • FIG. 1 is a diagram showing the relationship between the concentration of anti-Indofin mono- ⁇ antibody and absorbance in Example 1.
  • the immobilized antigen-antibody complex used in the present invention is produced by sequentially binding an anti-interfuron antibody and then interferon to a solid support.
  • the solid-phase carrier include plastic spheres, rods, plates, and the like made of plastic such as polystyrene and polypropylene, and a plastic plate is preferable.
  • anti-interferon antibody used in the present invention any anti-serum or antibody derived from animals obtained by administering interferon, or any monoclonal antibody produced by antibody-producing cells that cross-reacts with interferon can be used.
  • any of those commercially available as general reagents may be used.
  • human leukocyte fractions or cultured genetically modified lymphoblasts are used.
  • 9, Interferon-1 r obtained from the culture of T lymphocyte can be used.
  • a conventional method such as an adsorption method and a method using a cross-linking agent can be mentioned.
  • an anti-interferon antibody in an appropriate buffer is adsorbed to a solid phase carrier to solidify, and then the antigen, which is an antigen, is immobilized under the same buffer. What is necessary is just to couple
  • the amount of the anti-interferon antibody used in the solid phase immobilization reaction is preferably 5 to 500 U rd ⁇ , particularly preferably 20 to 300 UZ, and the amount of the interferon used is 100 to 500 U 0 0 UZm £, particularly 500 to 20000 UZm £, is preferred.
  • any buffer which is commonly used such as a phosphate buffer of pH 5 to 9, a Tris buffer, a Good buffer, and the like can be used as the buffer.
  • the blocking buffer used in the present invention is described in the report of Enzyme-mediatedimmu noassay, Plenum Preress, 1985, which is commonly used for the above buffer. Thus, those containing 1 to 3% of albumin or 0.05 to 0.1% of gelatin are preferred.
  • the quantification method of the present invention may be any of the competitive method and the Sandwich method as long as it is an immunological method using the above-described surface-conjugated antigen-antibody complex as a solid-phase carrier, and is particularly preferably ELISA. Therefore, in a preferred embodiment of the present invention, a test solution is reacted with the immobilized antigen-antibody complex, and after washing, a labeled secondary antibody is reacted, and the amount of the label bound to the solid phase is measured. And a method for quantifying the anti-interferon antibody and the interferon-anti-interfuron antibody complex in the test solution.
  • test liquid used in the present invention includes serum, plasma and the like.
  • the labeling agent for the labeled secondary antibody used in the quantification method of the present invention includes both RI and an enzyme, but an enzyme is preferred. Enzymes used for labeling include veroxidase, alkaline phosphatase, / 9-galactosidase and the like. Examples of the secondary antibody include a human IgG monoclonal antibody, a human IgG polyclonal antibody, and the like. Note that a commercially available labeled secondary antibody can also be used.
  • peroxidase is well known in addition to 0-phenylenediamine, which is well known as 2,2'-azino-di-di- (3-ethyl-pentylazolyl). 6) (ABTS), o-toluidine or 3-methyl-12-benzothiazoline hydrazone (MBTH) can be used.
  • ABTS o-toluidine or 3-methyl-12-benzothiazoline hydrazone
  • BC IP 5-bromo-4-chloro-3-indyl phosphite
  • the cleavage of 0- or p-nitrofuunyl) 9-D-galactosid ( ⁇ -, p-NPG) Free o- or p-nitrophenyl (0-, p-NP) can be used.
  • a reaction substrate for fluorescence tyramine or p-hydroxyphenylacetic acid or the like can be used for peroxidase, and fluorescein 1 / 9-D-galactoside or the like can be used for ⁇ -lauutosidase.
  • luminol or the like can be used for peroxidase.
  • a test solution is added to the immobilized antigen-antibody complex, and the mixture is incubated at room temperature to 40: 1 minute to 2 hours.
  • the solid phase is washed with a buffer, and a labeled secondary antibody is added, followed by incubating at room temperature to 40 for 1 minute to 2 hours.
  • add the substrate solution react at room temperature to 4 for 1 minute to 2 hours, add a reaction terminator, and measure the generated color, fluorescence or luminescence.
  • the amount of change in absorbance over a certain period of time may be measured.
  • an inorganic acid such as sulfuric acid or phosphoric acid, or an aqueous alkali solution such as sodium hydroxide can be used as a stop solution for the color-forming reaction.
  • an anti-interferon ⁇ -1 antibody was detected using a method based on Western plotting. That is, the interferon was electrophoresed, the membrane was subjected to plotting, and the diluted test serum was reacted. After washing, a peroxidase-labeled anti-human IgG was reacted and developed with the substrate diaminobenzidine. The test serum that developed color was determined to be an intestinal specimen. The results of the judgment are shown in Table 2 together with the results of the method of the present invention.
  • Table 2 shows that the method of the present invention has a good correlation with the results of the estan blotting method.
  • the method of the present invention comprises the steps of: Since interferon was sequentially bound, even if incompletely purified or interferon containing contaminants was used, accurate quantification was possible because it was purified in a face-phase system. In addition, since a carrier to which both an anti-interferon antibody and an interferon are bound is used as a solid phase, not only free anti-interferon sigma antibody present in the test solution but also interferon-interferon The antibody complex also reacted, and accurate measurement of anti-interfuron antibody became possible.

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Molecular Biology (AREA)
  • Engineering & Computer Science (AREA)
  • Cell Biology (AREA)
  • Chemical & Material Sciences (AREA)
  • Biomedical Technology (AREA)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Peptides Or Proteins (AREA)

Abstract

L'invention concerne une méthode de dosage d'un anticorps anti-interféron et d'un complexe interféron/anticorps anti-interféron présent dans un échantillon liquide, qui consiste à provoquer une réaction immunologique entre un antigène sur support solide et un échantillon liquide. Ledit antigène sur support solide comprend un complexe antigène sur support solide/anticorps préparé par fixation séquentielle d'un anticorps anti-interféron et d'un interféron sur un support en phase solide. La méthode selon l'invention permet la purification d'un interféron non purifié, même s'il a déjà été utilisé, au cours de l'étape de préparation de la phase solide, et le dosage rapide et précis d'un anticorps anti-interféron et d'un complexe interféron/anti-interféron présents dans le sérum.
PCT/JP1993/001657 1992-09-30 1993-11-12 Methode de dosage d'anticorps anti-interferon et reactif utilise dans ce dernier WO1995013539A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
JP4261627A JPH06109728A (ja) 1992-09-30 1992-09-30 抗インターフェロン抗体の定量法及びこれに用いる試薬
PCT/JP1993/001657 WO1995013539A1 (fr) 1992-09-30 1993-11-12 Methode de dosage d'anticorps anti-interferon et reactif utilise dans ce dernier

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP4261627A JPH06109728A (ja) 1992-09-30 1992-09-30 抗インターフェロン抗体の定量法及びこれに用いる試薬
PCT/JP1993/001657 WO1995013539A1 (fr) 1992-09-30 1993-11-12 Methode de dosage d'anticorps anti-interferon et reactif utilise dans ce dernier

Publications (1)

Publication Number Publication Date
WO1995013539A1 true WO1995013539A1 (fr) 1995-05-18

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7087726B2 (en) 2001-02-22 2006-08-08 Genentech, Inc. Anti-interferon-α antibodies
WO2013059299A1 (fr) * 2011-10-17 2013-04-25 The Uab Research Foundation Anticorps dirigés contre divers sous-types d'interféron, complexe ternaire interféron/récepteur d'interféron et utilisations associées

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS58147653A (ja) * 1982-02-08 1983-09-02 アボツト・ラボラトリ−ズ 種類特異性免疫グロブリン抗体の免疫分析法
JPS59501873A (ja) * 1982-08-09 1984-11-08 ブロック，マイロン・ジェイ 免疫検定装置及び方法

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS58147653A (ja) * 1982-02-08 1983-09-02 アボツト・ラボラトリ−ズ 種類特異性免疫グロブリン抗体の免疫分析法
JPS59501873A (ja) * 1982-08-09 1984-11-08 ブロック，マイロン・ジェイ 免疫検定装置及び方法

Non-Patent Citations (12)

* Cited by examiner, † Cited by third party
Title
BIOTECNOL. APL., Vol. 7, No. 2, 1990, S. CRUZ et al., "Quantification of Human Interferon alpha-2b Using Monoclonal Antibodies", pages 132-141. *
CHEM. PHARM. BULL., (TOKYO), Vol. 36, No. 5, 1988, N. KOBAYASHI et al., "A Solid-phase Enzyme Immunoassay Using Guinea-pig C3 to Detect Anti Mycoplasma-pulmonis Antibody in the Sera of Infected Rats", pages 1803-1807. *
CLIN. CHEM., Vol. 31, No. 2, 1985, Y-S, V. LIU et al., "A Monoclonal-antibody Enzyme Immunoassay for Detection of Hepatitis B Surface Antigen With Use of a Biotin-avidin System", pages 202-205. *
J. CLIN. MICROBIOL., Vol. 15, No. 3, 1982, M.M. COLLINS et al., "Solid Phase Complement C-1q Binding Fluorescence Immunoassay for Detection of Circulating Immune Complexes", pages 456-464. *
J. CLIN. MICROBIOL., Vol. 17, No. 3, 1983, S. GIUNTA et al., "Competitive Enzyme Immunoassay for Detection of Complement Fixing Antibodies in Diagnostic Virology", pages 507-510. *
J. IMMUNOL. METHODS, Vol. 116, No. 2, 1989, T. JITSUKAWA et al., "Increased Coating Efficiency of Antigens and Preservation of Original Antigentic Structure After Coating in ELISA", pages 251-7. *
J. IMMUNOL. METHODS, Vol. 119, No. 1, 1989, M.L. OVERALL et al., "Comparison of Different ELISAs for the Detection of Monoclonal Antibodies to Human Interferon-alpha", pages 27-33. *
J. IMMUNOL. METHODS, Vol. 128, No. 1, 1990, T. TAGUCHI et al., "Detection of Individual Mouse Splentic T Cells Producing IFN-gamma and IL-5 Using the Enzyme-linked Immunospot (ELISPOT) Assay", pages 65-73. *
J. IMMUNOL. METHODS, Vol. 130, No. 2, 1990, O. PRUMMER et al., "Filter Spot-Elisa for the Enumeration of Interferon-alpha Antibody-secreting Cells", pages 187-93. *
J. IMMUNOL. METHODS, Vol. 158, No. 2, 1993, P.K.M. NGAI et al., "Protein A Antibody-capture ELISA PACE an ELISA Format to Avoid Denaturation of Surface-adsorbed Antigens", pages 267-276. *
RES. DISCL., Vol. 256, (1985-8), ANON., "Enzyme-immunological Determination of Interferon-specific Antibodies in Physiological Fluids Using the Double Antigen Sandwich Technique According to the One-step Test Method", page 389. *
RES. DISCL., Vol. 256, (1985-8), ANON., "Enzyme-immunological Test for the Determination of Interferon Antibodies in Physiological Fluids According to the Double Antigen Sandwich Technique", page 397. *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7087726B2 (en) 2001-02-22 2006-08-08 Genentech, Inc. Anti-interferon-α antibodies
WO2013059299A1 (fr) * 2011-10-17 2013-04-25 The Uab Research Foundation Anticorps dirigés contre divers sous-types d'interféron, complexe ternaire interféron/récepteur d'interféron et utilisations associées

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