WO1990013817A1 - PROCEDE DE DOSAGE D'ANTICORPS IgG ANTI-ALLERGENES SPECIFIQUES, FRACTION RICHE EN IgG4 OBTENUE SELON CE PROCEDE ET COMPOSITION PHARMACEUTIQUE CONTENANT CETTE FRACTION - Google Patents
PROCEDE DE DOSAGE D'ANTICORPS IgG ANTI-ALLERGENES SPECIFIQUES, FRACTION RICHE EN IgG4 OBTENUE SELON CE PROCEDE ET COMPOSITION PHARMACEUTIQUE CONTENANT CETTE FRACTION Download PDFInfo
- Publication number
- WO1990013817A1 WO1990013817A1 PCT/FR1990/000335 FR9000335W WO9013817A1 WO 1990013817 A1 WO1990013817 A1 WO 1990013817A1 FR 9000335 W FR9000335 W FR 9000335W WO 9013817 A1 WO9013817 A1 WO 9013817A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- igg
- fraction
- igg4
- specific
- allergen
- Prior art date
Links
- 238000000034 method Methods 0.000 title claims abstract description 42
- 239000008194 pharmaceutical composition Substances 0.000 title claims abstract description 5
- 239000013566 allergen Substances 0.000 claims abstract description 18
- 238000002965 ELISA Methods 0.000 claims abstract description 7
- 210000002966 serum Anatomy 0.000 claims description 11
- 108060003951 Immunoglobulin Proteins 0.000 claims description 10
- 102000018358 immunoglobulin Human genes 0.000 claims description 10
- 238000000746 purification Methods 0.000 claims description 9
- WWZKQHOCKIZLMA-UHFFFAOYSA-N octanoic acid Chemical compound CCCCCCCC(O)=O WWZKQHOCKIZLMA-UHFFFAOYSA-N 0.000 claims description 8
- 238000003556 assay Methods 0.000 claims description 6
- 239000002244 precipitate Substances 0.000 claims description 5
- 239000005635 Caprylic acid (CAS 124-07-2) Substances 0.000 claims description 4
- 229960002446 octanoic acid Drugs 0.000 claims description 4
- 230000008030 elimination Effects 0.000 claims description 2
- 238000003379 elimination reaction Methods 0.000 claims description 2
- 229940043517 specific immunoglobulins Drugs 0.000 claims description 2
- 238000002360 preparation method Methods 0.000 claims 1
- 229960004784 allergens Drugs 0.000 description 4
- 238000010790 dilution Methods 0.000 description 4
- 239000012895 dilution Substances 0.000 description 4
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 4
- 239000000284 extract Substances 0.000 description 4
- 229940072221 immunoglobulins Drugs 0.000 description 4
- 239000002953 phosphate buffered saline Substances 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 238000011282 treatment Methods 0.000 description 4
- 230000000172 allergic effect Effects 0.000 description 3
- 239000000427 antigen Substances 0.000 description 3
- 102000036639 antigens Human genes 0.000 description 3
- 108091007433 antigens Proteins 0.000 description 3
- 208000010668 atopic eczema Diseases 0.000 description 3
- 229940046528 grass pollen Drugs 0.000 description 3
- 238000011534 incubation Methods 0.000 description 3
- 210000002381 plasma Anatomy 0.000 description 3
- 102000009027 Albumins Human genes 0.000 description 2
- 108010088751 Albumins Proteins 0.000 description 2
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 2
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 2
- 240000004585 Dactylis glomerata Species 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 241000209504 Poaceae Species 0.000 description 2
- 230000009102 absorption Effects 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 238000000586 desensitisation Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 230000003287 optical effect Effects 0.000 description 2
- 238000002616 plasmapheresis Methods 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- XZKIHKMTEMTJQX-UHFFFAOYSA-N 4-Nitrophenyl Phosphate Chemical compound OP(O)(=O)OC1=CC=C([N+]([O-])=O)C=C1 XZKIHKMTEMTJQX-UHFFFAOYSA-N 0.000 description 1
- 206010002368 Anger Diseases 0.000 description 1
- 244000025254 Cannabis sativa Species 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 241000238710 Dermatophagoides Species 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 229920001213 Polysorbate 20 Polymers 0.000 description 1
- 229920002684 Sepharose Polymers 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 230000001476 alcoholic effect Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 230000037029 cross reaction Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005194 fractionation Methods 0.000 description 1
- 230000000521 hyperimmunizing effect Effects 0.000 description 1
- 238000003018 immunoassay Methods 0.000 description 1
- 229940027941 immunoglobulin g Drugs 0.000 description 1
- 238000010348 incorporation Methods 0.000 description 1
- 238000005497 microtitration Methods 0.000 description 1
- 229940062713 mite extract Drugs 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 1
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
- 229920000136 polysorbate Polymers 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000002285 radioactive effect Effects 0.000 description 1
- 238000003127 radioimmunoassay Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
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
- G01N33/6854—Immunoglobulins
-
- 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 in particular to the assay of anti-allergenic antibodies and to a method making it possible to obtain immunogiobulins having a high titer in specific anti-allergenic IgGs, in particular anti-grass pollen.
- European patent n ° 86 901887.9 describes a process making it possible to obtain immunoglobulins having a high titer in specific IgG anti-allergens.
- One of the drawbacks of this method is that it must start from the serum of patients whose content of specific anti-allergen IgG is most unknown time. This is all the less the case since the sera treated are mostly mixtures from different donors.
- the method according to the present invention is therefore a method for determining the content of specific IgG allergens contained in a medium, characterized in that the ELISA technique is used, in which the IgGs contained in the sample of the medium are fixed with support via the aergergene, the specific allergen IgGs are detected by means of a non-human monoclonal anti-human IgG ⁇ antibody, the presence of which is revealed by a labeled antibody corresponding to the non-human epitope part of the monclonal antibody.
- pollen Among the more particularly important allergens to be detected by the method of the present invention, mention should be made of pollen, pollen extracts from trees or grasses and dermatophagoid mites farinae and pteronyssinus.
- this ELISA technique is applied in an original manner to allergens of the pollen and mite extract type and uses anti-IgG monoclonal antibodies which allow the detection of specific IgGs.
- the ELISA technique according to the invention does not have the aforementioned drawbacks and makes it possible to express the results in absolute value (mg / ml).
- the present invention relates more particularly to the incorporation of this assay method into a method for obtaining immunogiobulins anti-allergen hyperimmune, method in which, prior to the purification of the sera by any suitable method, the donor serum is assayed and the sera having specific anti-allergen IgG antibody immunoglobulin levels of at least 0.2 ⁇ g / ml and preferably at least minus 1 ⁇ g / l.
- the purification methods which can be used mention must be made of the purification method mentioned at the beginning of this description, that is to say a method which, in the fractions I + II + III of Cohn, precipitates the specific immunoglobulins thanks to caprylic acid after re-solution of the precipitate I + II + III.
- the present invention can be implemented according to the method which comprises the following steps: a) assaying the serum samples according to the invention; b) elimination of the samples having an immunoglobulin anti-allergen specific IgG antibody content of less than 0.2 ⁇ g / ml; c) purification of the serum samples to obtain said fraction rich in IgG.
- the present invention also relates to a fraction rich in IgG, which can be obtained by the above method.
- the present invention further relates to a pharmaceutical composition which can be used in desensitization treatments containing the fraction rich in IgG referred to above.
- Example 1 The examples below will better illustrate certain characteristics of the assay and purification process according to the invention.
- Example 1 The examples below will better illustrate certain characteristics of the assay and purification process according to the invention.
- the monoclonal anti-human IgG antibodies come from Merck, Nogent-sur-Marne antibodies and immunsera,
- the goat anti-immunoglobulin G mouse antiserum conjugated with alkaline phosphatase comes from Laboratoires Sigma Chemical Co. (Saint-Louis, USA). It was absorbed by human IgG to avoid cross-reactions with human IgG. Immuno-enzymatic technique for the measurement of grass pollen IgG
- 0.1 ml of a solution of soluble pollen antigens (Dactylis Glomerata) at 2 ⁇ g / ml in phosphate saline buffer pH 7 are deposited in the wells of the plates. microtitration. After an incubation of 18 hours at ° C, the wells are washed 5 times in phosphate-buffered saline,
- the results are expressed in optical density (OD).
- OD optical density
- 12,500 phasmapheresis donors were screened for subjects with high grass anti-polain IgG antibody levels.
- the measurements were carried out at a single dilution of the 50th and expressed in optical density as described in Example 1. This screening was carried out either from subjects selected by an interrogation relating to the existence of allergic history, or subjects randomly selected from the population of plasmapheresis donors.
- the plasmas selected have been "pooled".
- the rate of specific anti-pollen IgG antibodies is 7 ⁇ g / ml of specific IgG4, measured by ELISA, in the “pool” of plasmas.
- the immunoglobulins were fractionated by a technique using the ethanol and caprylic acid fraction.
- the alcoholic fraction II + III is precipitated (precipitate A from the Kitler and Nisehman fractionation scheme) according to a method using caprylic acid.
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Analytical Chemistry (AREA)
- Biotechnology (AREA)
- Pathology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Cell Biology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Peptides Or Proteins (AREA)
Abstract
Description
Claims
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
FR8906281A FR2646917A1 (fr) | 1989-05-12 | 1989-05-12 | Procede de dosage d'anti-corps igg anti-allergenes specifiques, fraction riche en igg4 obtenue selon ce procede et composition pharmaceutique contenant cette fraction |
FR89/06281 | 1989-05-12 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO1990013817A1 true WO1990013817A1 (fr) | 1990-11-15 |
Family
ID=9381627
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/FR1990/000335 WO1990013817A1 (fr) | 1989-05-12 | 1990-05-11 | PROCEDE DE DOSAGE D'ANTICORPS IgG ANTI-ALLERGENES SPECIFIQUES, FRACTION RICHE EN IgG4 OBTENUE SELON CE PROCEDE ET COMPOSITION PHARMACEUTIQUE CONTENANT CETTE FRACTION |
Country Status (3)
Country | Link |
---|---|
FR (1) | FR2646917A1 (fr) |
IT (1) | IT9020295A1 (fr) |
WO (1) | WO1990013817A1 (fr) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5367054A (en) * | 1993-04-12 | 1994-11-22 | Stolle Research & Development Corp. | Large-scale purification of egg immunoglobulin |
EP2232265A1 (fr) * | 2008-01-02 | 2010-09-29 | Fundacao De Amparo A Pesquisa Do Estado De Minas Gerais - Fapemig | Procédé et kit pour quantifier des sous-classes d'igg humaines spécifiques d'allergènes pour le contrôle et le soin d'immunothérapie spécifique |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0161868A2 (fr) * | 1984-05-11 | 1985-11-21 | BioWhittaker, Inc. | Dosage fluorimétrique d'IgG4 |
EP0163141A2 (fr) * | 1984-04-27 | 1985-12-04 | Shionogi & Co., Ltd. | Anticorps monoclonal anti-IgG humain et son procédé de préparation |
WO1986005099A1 (fr) * | 1985-03-07 | 1986-09-12 | Centre National De Transfusion Sanguine | Preparations d'immunoglobulines presentant des titres eleves en anticorps bloquants |
EP0205352A2 (fr) * | 1985-06-12 | 1986-12-17 | Shionogi & Co., Ltd. | Anticorps IgG4-anti humain monoclonaux, leur préparation et utilisation |
-
1989
- 1989-05-12 FR FR8906281A patent/FR2646917A1/fr not_active Withdrawn
-
1990
- 1990-05-11 WO PCT/FR1990/000335 patent/WO1990013817A1/fr not_active Application Discontinuation
- 1990-05-14 IT IT020295A patent/IT9020295A1/it not_active Application Discontinuation
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0163141A2 (fr) * | 1984-04-27 | 1985-12-04 | Shionogi & Co., Ltd. | Anticorps monoclonal anti-IgG humain et son procédé de préparation |
EP0161868A2 (fr) * | 1984-05-11 | 1985-11-21 | BioWhittaker, Inc. | Dosage fluorimétrique d'IgG4 |
WO1986005099A1 (fr) * | 1985-03-07 | 1986-09-12 | Centre National De Transfusion Sanguine | Preparations d'immunoglobulines presentant des titres eleves en anticorps bloquants |
EP0205352A2 (fr) * | 1985-06-12 | 1986-12-17 | Shionogi & Co., Ltd. | Anticorps IgG4-anti humain monoclonaux, leur préparation et utilisation |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5367054A (en) * | 1993-04-12 | 1994-11-22 | Stolle Research & Development Corp. | Large-scale purification of egg immunoglobulin |
EP2232265A1 (fr) * | 2008-01-02 | 2010-09-29 | Fundacao De Amparo A Pesquisa Do Estado De Minas Gerais - Fapemig | Procédé et kit pour quantifier des sous-classes d'igg humaines spécifiques d'allergènes pour le contrôle et le soin d'immunothérapie spécifique |
EP2232265A4 (fr) * | 2008-01-02 | 2010-12-29 | Fundacao De Amparo A Pesquisa | Procédé et kit pour quantifier des sous-classes d'igg humaines spécifiques d'allergènes pour le contrôle et le soin d'immunothérapie spécifique |
Also Published As
Publication number | Publication date |
---|---|
IT9020295A1 (it) | 1990-11-13 |
FR2646917A1 (fr) | 1990-11-16 |
IT9020295A0 (it) | 1990-05-14 |
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