WO2000040967A2 - Verfahren und vorrichtung zur diagnose allergischer symptome - Google Patents
Verfahren und vorrichtung zur diagnose allergischer symptome Download PDFInfo
- Publication number
- WO2000040967A2 WO2000040967A2 PCT/DE2000/000001 DE0000001W WO0040967A2 WO 2000040967 A2 WO2000040967 A2 WO 2000040967A2 DE 0000001 W DE0000001 W DE 0000001W WO 0040967 A2 WO0040967 A2 WO 0040967A2
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- antibodies
- binding protein
- protein
- carrier
- patient
- Prior art date
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54366—Apparatus specially adapted for solid-phase testing
- G01N33/54373—Apparatus specially adapted for solid-phase testing involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
-
- 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
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
- B01J2219/00583—Features relative to the processes being carried out
- B01J2219/00603—Making arrays on substantially continuous surfaces
- B01J2219/00605—Making arrays on substantially continuous surfaces the compounds being directly bound or immobilised to solid supports
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
- B01J2219/00583—Features relative to the processes being carried out
- B01J2219/00603—Making arrays on substantially continuous surfaces
- B01J2219/00605—Making arrays on substantially continuous surfaces the compounds being directly bound or immobilised to solid supports
- B01J2219/00612—Making arrays on substantially continuous surfaces the compounds being directly bound or immobilised to solid supports the surface being inorganic
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
- B01J2219/00583—Features relative to the processes being carried out
- B01J2219/00603—Making arrays on substantially continuous surfaces
- B01J2219/00605—Making arrays on substantially continuous surfaces the compounds being directly bound or immobilised to solid supports
- B01J2219/00614—Delimitation of the attachment areas
- B01J2219/00617—Delimitation of the attachment areas by chemical means
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
- B01J2219/00583—Features relative to the processes being carried out
- B01J2219/00603—Making arrays on substantially continuous surfaces
- B01J2219/00605—Making arrays on substantially continuous surfaces the compounds being directly bound or immobilised to solid supports
- B01J2219/00623—Immobilisation or binding
- B01J2219/0063—Other, e.g. van der Waals forces, hydrogen bonding
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
- B01J2219/00583—Features relative to the processes being carried out
- B01J2219/00603—Making arrays on substantially continuous surfaces
- B01J2219/00605—Making arrays on substantially continuous surfaces the compounds being directly bound or immobilised to solid supports
- B01J2219/00632—Introduction of reactive groups to the surface
- B01J2219/00637—Introduction of reactive groups to the surface by coating it with another layer
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
- B01J2219/00583—Features relative to the processes being carried out
- B01J2219/00603—Making arrays on substantially continuous surfaces
- B01J2219/00659—Two-dimensional arrays
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
- B01J2219/00718—Type of compounds synthesised
- B01J2219/0072—Organic compounds
Definitions
- the invention relates to a method for diagnosing allergic symptoms by detecting in vitro immune reactions, in which a binding protein is applied to a carrier, immobilized on the antibody of a patient to be examined by coupling to the binding protein and then exposed to an antigen solution which triggers the allergy Contains antigens.
- the invention further relates to a device for diagnosing allergic symptoms by detecting in vitro immune reactions.
- EP 0 629 857 A 2 discloses a method for immunochemical determination of antigens.
- a binding protein on which primary antibodies are immobilized is applied to the surface of a support.
- Antigens to be determined are bound to the primary antibodies and thus also immobilized.
- the antigens are determined by determining an immune reaction with labeled secondary antibodies.
- a disadvantage of this method is that, in addition to the primary antibodies, labeled secondary antibodies are necessary.
- the antigens, which entered into a complex with the immobilized specific antibodies in the previous process step, can in the subsequent incubation steps in a backward reaction partially detach from the complex and thus elude the detection reaction, which, among other things, severely limits the sensitivity. It is therefore an object of the present invention to find a method in which labeled secondary antibodies can be dispensed with.
- the carrier is designed as a reflective solid carrier on which the binding protein is applied in a monomolecular, surface-active layer, and that after an incubation period, an immunoreaction between the antibody and the antigen caused increase in layer thickness of the protein-antibody layer is determined by an ellipsometric measurement method.
- protein A is applied as the binding protein by means of the known Langmuir-Blodgett technique.
- protein G or other suitable proteins as the binding protein and to apply it using the same technique.
- the binding protein in a desired monomolecular surface-active layer.
- the monomolecular layer ensures that minimal substances can be used and detected.
- a silicon wafer is used as the carrier.
- silicon as the carrier material, a correspondingly high resolution can be achieved for the ellipsometric determination of the layer ceiling because of the high refractive index of silicon.
- the carrier is at least partially provided with a lattice-shaped cover before the binding protein is applied. After the protein has been applied in islands separated from one another by the cover grid, the cover grid is removed.
- Immunoglobulin antibody used. For example, the most frequently occurring immunoglobulin G in the immune system
- IgG immunoglobulin E antibodies
- IgE immunoglobulin E antibodies
- the antibody (IgG) - like a "Y" - consists of three areas. The Fab regions, corresponding to the poor, are responsible for the immune-specific responsiveness, while the Fc fragment, the foot end, reacts unspecifically.
- the IgG is coupled to the binding protein, for example protein A, via the Fc fragment.
- isotypic antibodies separated from patient serum are used.
- isotypic antibodies can also be separated from blood, urine or other body fluids.
- the separated antibodies are applied by micropipetting to the binding protein arranged in separate islands.
- the islands of the binding protein can easily be populated by serial micropipetting or by a complete micro-pipetting that is raster-synchronous and corresponds to the island matrix.
- the carrier is scanned on a two-coordinate measuring table of an egg lipometer in a software-controlled manner by the laser beam of the egg lipometer. The result of the scanning process is then evaluated using an evaluation software.
- the software-controlled ellipsometric measurement of the layer thickness increase enables the ellipsometric layer thickness determination to be carried out largely automatically and error-free in a time-saving manner.
- a mass separator is used for a quantitative diagnosis, which breaks down the substance of the islands on which an immune reaction has taken place into their atomic components.
- the separated elementary fractions obtained are reduced by the known protein and antibody proportions, so that the residual substance results in a sought-after, allergy-causing factor.
- the allergenic factor is compared with data stored in an allergen database and evaluated.
- the separator result can be reconstructed using the allergen database - a software library - and identified by comparisons and restrictions.
- the method according to the invention is thus quantitatively much more precise than the previously known method.
- the method according to the invention can be expanded to a generally applicable method.
- EP 629 857 A 2 has the disadvantages described and is not suitable for optical measurement in an egg lipometer.
- Another object of the present invention is therefore to provide a device for diagnosing allergic symptoms by detecting in vitro immune reactions, which does not require labeled secondary antibodies and is suitable for non-destructive optical measurement.
- a sensor base is formed from a reflective solid support on which a monomolecular layer of a surface-active binding protein is applied in islands in a matrix, to which patient-specific antibodies can be attached, that can react with antigens contained in a solution, and that the immune reactions can be measured in an egg lipometer.
- the sensor base is formed from a reflective carrier, on the islands of a surface-active binding protein, to which patient-specific antibodies, which can react with antigens contained in a solution, can be attached, it is possible to mount the device or the carrier in an egg lipometer evaluate so that the immune reactions can be measured in an egg lipometer.
- the reflective carrier is made of silicon.
- silicon By using silicon, a high resolution is achieved for the ellipsometric determination of the layer thickness because of the high refractive index of silicon.
- protein A is used as the binding protein.
- Immunoglobulin G antibodies IgG of the patient to be examined are used as antibodies.
- the Fc receptor of protein A binds the IgG with its Fc region to protein A.
- FIG. 1 a schematic plan view of a carrier in an enlarged view
- FIG. 2 shows a plan view of the carrier from FIG. 1 with a schematic illustration of a micropipette arranged in a row
- FIG. 3 a basic illustration, not to scale, of a side view of a carrier
- Figure 4 is a schematic side view of a carrier inserted into a tub and
- Figure 5 a simplified flow diagram of a method for the detection of antigen.
- a device for diagnosing allergic symptoms essentially consists of a sensor base 1 to which patient-specific allergens 2 can be connected.
- the sensor base 1 essentially consists of a carrier 3, a binding protein 4 and patient-specific antibodies 5.
- the carrier 3 is designed as a rectangular silicon plate on which a monomolecular layer of a binding protein 4 is arranged using the Langmuir-Blodgett technique (coating of monomolecular films of surface-active substances by means of a special dipping process).
- Protein A PrA
- protein G protein
- the binding protein 4 is arranged in a matrix in islands 8 on the Si plate.
- the carrier 3 has an exposed edge 6.
- the released edge 6 serves on the one hand as an attack surface for holders and, on the other hand, optionally as a reference surface for determining the refractive index of the carrier 3.
- the islands 8 are equipped with patient-specific antibodies 5. Different islands 8 have antibodies 5, each with a different specificity.
- Isotypic immunoglobulin G antibodies (IgG) separated from patient serum are used as antibody 5.
- immunoglobulin E antibodies IgE.
- a cover grid 9 made of a wax-like material is applied to the carrier 3 or the Si plate.
- the interstices left free by the cover grid 9 are monomolecularly coated with protein A by dipping, so that islands 8 made of PrA are formed by the cover grid 9.
- the antibodies (IgG) 5 separated from patient sera are applied to the patient with the help of micropipettes 10 arranged in a row. be 8 applied.
- the sensor base 1 is populated with the number of a raster sequence (corresponds to a row in the carrier grid, for example four islands 8) on micropipettes 10, the micropipettes 10 being drawn sequentially over the sensor base 1 or the carrier 3 while the carrier 3 a matrix width 7 is pushed further. For example, with four islands 8 arranged in a row, the matrix width 7 corresponds to four times the island width 11.
- a micropipetting grid corresponding to the carrier grid is produced, which is possible, for example, by micro-etching Si platelets of appropriate thickness using its crystal structure. With the help of the micropipetting grid, a grid-synchronized complete assembly is made possible.
- the islands 8 are freed of excess antibodies by rinsing.
- the cover grid 9 is then removed from the carrier.
- the sensor base 1 which now acts as a biosensor, is exposed in a tub 12 to an antigen or allergen solution 13 with patient-specific antigens or allergens 2.
- the increase in layer thickness to be expected when the antigens or allergens 2 are bound (immune reaction) to the antibodies 5 is then detected using an egg lipometer (not shown).
- the carrier 3 is scanned in a software-controlled manner on a two-coordinate measuring table of the egg lipometer by the laser beam of the egg lipometer. The result of the scanning process is then evaluated using an evaluation software.
- the substance of the islands 8, on which an immune reaction has taken place is broken down into its atomic components.
- the separated elementary fractions achieved are reduced by the known binding protein and antibody proportions.
- the residual substance is the allergy-causing factor that is being sought and is evaluated in an allergen database or software library.
- This allergen database is a collection of all known allergens with the distribution of their elementary components, their biochemical structure as well as molecular structural elements and all conditions necessary for the evaluation.
- the separator result is reconstructed with the help of the allergen database using an integrated processing program and identified by comparisons and restrictions.
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- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Cell Biology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Microbiology (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Investigating Or Analysing Materials By Optical Means (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
Description
Claims
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2000592635A JP2002534682A (ja) | 1999-01-06 | 2000-01-03 | アレルギー症状を診断する方法と装置 |
EP00904801A EP1145008A2 (de) | 1999-01-06 | 2000-01-03 | Verfahren und vorrichtung zur diagnose allergischer symptome |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE1999100119 DE19900119C2 (de) | 1999-01-06 | 1999-01-06 | Verfahren und Vorrichtung zur Diagnose allergischer Symptome |
DE19900119.7 | 1999-01-06 |
Publications (2)
Publication Number | Publication Date |
---|---|
WO2000040967A2 true WO2000040967A2 (de) | 2000-07-13 |
WO2000040967A3 WO2000040967A3 (de) | 2001-10-25 |
Family
ID=7893600
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/DE2000/000001 WO2000040967A2 (de) | 1999-01-06 | 2000-01-03 | Verfahren und vorrichtung zur diagnose allergischer symptome |
Country Status (4)
Country | Link |
---|---|
EP (1) | EP1145008A2 (de) |
JP (1) | JP2002534682A (de) |
DE (1) | DE19900119C2 (de) |
WO (1) | WO2000040967A2 (de) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP2282206A2 (de) | 2004-02-17 | 2011-02-09 | DST Diagnostische Systeme & Technologien GmbH | Verfahren und Vorrichtung zur Bestimmung mehrerer Analyten mit simultaner internen Kontrolle in einer grafischen Kombination |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE10120562C2 (de) * | 2001-04-26 | 2003-04-10 | Horst Messer | Verfahren zur Diagnose von übertragbaren spongiformen Enzephalopathien |
Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1992016286A1 (en) * | 1991-03-22 | 1992-10-01 | Göteborgs Analyslaboratorium Ab | Process and means for mixing at least two components in a flexible tube |
US5552272A (en) * | 1993-06-10 | 1996-09-03 | Biostar, Inc. | Detection of an analyte by fluorescence using a thin film optical device |
US5633724A (en) * | 1995-08-29 | 1997-05-27 | Hewlett-Packard Company | Evanescent scanning of biochemical array |
US5812272A (en) * | 1997-01-30 | 1998-09-22 | Hewlett-Packard Company | Apparatus and method with tiled light source array for integrated assay sensing |
EP0886141A1 (de) * | 1997-06-23 | 1998-12-23 | C.S.E.M. Centre Suisse D'electronique Et De Microtechnique Sa | Optische Sensorvorrichtung und Verfahren zur hochempfindlichen Detektion von chemischen oder biochemischen Analyten |
Family Cites Families (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE3025022C2 (de) * | 1980-07-02 | 1985-07-04 | Wang, Wei-Kung, T'ai-pei | Verfahren zur Bestimmung biologischer Teilchen durch induzierte Signale |
EP0067921B1 (de) * | 1981-06-22 | 1987-11-11 | Prutec Limited | Verfahren zum Bestimmen bioaktiver Substanzen |
SE9602545L (sv) * | 1996-06-25 | 1997-12-26 | Michael Mecklenburg | Metod för att diskriminera komplexa biologiska prover |
DE19629243A1 (de) * | 1996-07-19 | 1998-01-29 | Udo Dr Ris | Verfahren zur quantitativen und/oder qualitativen Bestimmung von Atomen oder Molekülen |
-
1999
- 1999-01-06 DE DE1999100119 patent/DE19900119C2/de not_active Expired - Fee Related
-
2000
- 2000-01-03 WO PCT/DE2000/000001 patent/WO2000040967A2/de not_active Application Discontinuation
- 2000-01-03 JP JP2000592635A patent/JP2002534682A/ja not_active Withdrawn
- 2000-01-03 EP EP00904801A patent/EP1145008A2/de not_active Withdrawn
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1992016286A1 (en) * | 1991-03-22 | 1992-10-01 | Göteborgs Analyslaboratorium Ab | Process and means for mixing at least two components in a flexible tube |
US5552272A (en) * | 1993-06-10 | 1996-09-03 | Biostar, Inc. | Detection of an analyte by fluorescence using a thin film optical device |
US5633724A (en) * | 1995-08-29 | 1997-05-27 | Hewlett-Packard Company | Evanescent scanning of biochemical array |
US5812272A (en) * | 1997-01-30 | 1998-09-22 | Hewlett-Packard Company | Apparatus and method with tiled light source array for integrated assay sensing |
EP0886141A1 (de) * | 1997-06-23 | 1998-12-23 | C.S.E.M. Centre Suisse D'electronique Et De Microtechnique Sa | Optische Sensorvorrichtung und Verfahren zur hochempfindlichen Detektion von chemischen oder biochemischen Analyten |
Non-Patent Citations (3)
Title |
---|
LIN V S -Y ET AL: "A POROUS SILICON-BASED OPTICAL INTERFEROMETRIC BIOSENSOR" SCIENCE,US,AMERICAN ASSOCIATION FOR THE ADVANCEMENT OF SCIENCE,, Bd. 278, Nr. 5339, 31. Oktober 1997 (1997-10-31), Seiten 840-843, XP000876896 ISSN: 0036-8075 * |
OSTROFF R.M. ET AL.: "Thin film biosensor for rapid visual detection of nucleic acid targets" CLIN. CHEM., Bd. 45, Nr. 9, 1999, Seiten 1659-1664, XP002140865 * |
SILZEL J.W. ET AL.: "Mass-sensing, multianalyte microarray immunoassay with imaging detection" CLIN. CHEM., Bd. 44, Nr. 9, 1998, Seiten 2036-2043, XP002140864 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP2282206A2 (de) | 2004-02-17 | 2011-02-09 | DST Diagnostische Systeme & Technologien GmbH | Verfahren und Vorrichtung zur Bestimmung mehrerer Analyten mit simultaner internen Kontrolle in einer grafischen Kombination |
Also Published As
Publication number | Publication date |
---|---|
EP1145008A2 (de) | 2001-10-17 |
DE19900119A1 (de) | 2000-08-17 |
DE19900119C2 (de) | 2001-02-22 |
JP2002534682A (ja) | 2002-10-15 |
WO2000040967A3 (de) | 2001-10-25 |
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