WO2002086511A2 - VERFAHREN ZUR UNTERSUCHUNG VON PRION-PROTEIN ENTHALTENDEN PROBEN AUF DAS EVENTUELLE VORLIEGEN DER PrPSc-FORM - Google Patents
VERFAHREN ZUR UNTERSUCHUNG VON PRION-PROTEIN ENTHALTENDEN PROBEN AUF DAS EVENTUELLE VORLIEGEN DER PrPSc-FORM Download PDFInfo
- Publication number
- WO2002086511A2 WO2002086511A2 PCT/EP2002/004341 EP0204341W WO02086511A2 WO 2002086511 A2 WO2002086511 A2 WO 2002086511A2 EP 0204341 W EP0204341 W EP 0204341W WO 02086511 A2 WO02086511 A2 WO 02086511A2
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- prp
- sample
- prion protein
- protease
- molecules
- 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
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
- G01N33/6896—Neurological disorders, e.g. Alzheimer's disease
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/28—Neurological disorders
- G01N2800/2814—Dementia; Cognitive disorders
- G01N2800/2828—Prion diseases
Definitions
- the invention relates to a method according to the preamble of claim 1.
- Generic methods are currently especially in mammalian screening studies, e.g. Slaughter cattle, applied to communicable, degenerative neurological diseases.
- Such diseases which are summarized under the term spongiform encephalopathies or prion diseases, occur as BSE in cattle and e.g. as scrapies in sheep, or as Kuru or Creuzfeldt-Jakob disease in humans.
- prion diseases are communicable, although the infectiousness has not yet been fully clarified.
- the only molecule associated with the infectious agent has so far been found to be a disease-specific prion protein (PrP c ) which is an abnormal isoform of a normal mammalian protein (PrP c ) of unknown function.
- PrP c disease-specific prion protein
- Both isoforms PrP Sc and PrP c are the same in terms of molecular weight and amino acid sequence. They differ in their spatial folding.
- PrP Sc proteins are able to convert normal PrP c proteins into disease-specific folding, which would explain the infectivity of PrP Sc proteins.
- PrP Sc the central disease molecule
- samples from infected sources contain not only PrP Sc but also prion protein in the PrP c form. During the process, a differentiation must therefore be made between the PrP c and the possibly existing PrP Sc form.
- PrP c form is completely protease digestible, while in the PrP c form only one C-terminal region is protease sensitive and one region of the prion protein protease, designated as PrP 27-30, is resistant is.
- the sample to be examined is therefore first digested with a protease in a first step (step a), it being assumed that after digestion, no protease-sensitive areas of the prion protein are present in the sample and only in the case of infectious sample material the protease resistant region PrP 27-30 of the PrP Sc form remains. Accordingly, after the digestion, in a further step (step b) it is only checked whether the area PrP 27-30 can be detected in the sample or not. For example, specific antibodies binding in the range PrP 27-30 are used for detection. Any antibody-PrP 27-30 complexes formed are then detected using conventional methods, for example Elisa tests (Moynagh and Schimmel; Nature 1999 Jul 8; 400 (6470): 105). If the proof is positive, ie if antibody-PrP 27-30 complexes can be detected, it is assumed that the sample contains PrP Sc and that the organism from which the sample comes was infected.
- the object of the invention is to develop generic methods so that a more reliable statement is possible.
- the sample in step b is not only checked to determine whether it contains the PrP 27-30 region, but also to check whether protease is still present in the sample. sensitive areas of the prion protein are contained.
- the method according to the invention thus allows a statement about the possible presence or absence of PrP 27-30 in the digested sample and a statement as to whether the digestion was complete or not.
- PrP 27-30 in a digested sample is only considered as an indication of PrP Sc if none of the digested sample is present at the same time Protease sensitive areas of the prion protein are more detectable. If such protease sensitive areas can still be detected in the sample even after digestion, the possible detection of PrP 27-30 is not a clear indication of PrP Sc but could also mean that a corresponding area of the PrP c form was not completely digested. In this case the sample would have to be examined again with, for example, higher protease concentrations or longer digestion times.
- false positive examination results can be excluded in a particularly safe and simple manner.
- the significance of a test essentially depends on the fact that false positive results are minimized.
- PrP 27-30 or protease-sensitive regions of the prion protein are detected by means of molecules which bind specifically to the prion protein in the respective regions and which are described below as molecule A (specific for a protease-sensitive) Area) and molecule B (specifically for the area PrP 27-30).
- a common method provides that in step a the digestion of the sample takes place and in step b the molecules A and B of the digested sample are added and it is checked whether there are complexes from the prion protein with the molecule in the sample A and / or B have formed. The evaluation is then carried out depending on whether and which complexes have formed.
- the sample contains PrP Sc . Are there additional complexes that contain the molecule A there is a risk of a false positive result. If no complexes or only complexes with molecule A are found, the sample is negative.
- Molecules A and B can in particular be antibodies (hereinafter referred to as antibodies A and B) which specifically recognize the corresponding regions of the prion protein.
- antibodies A and B antibodies which specifically recognize the corresponding regions of the prion protein.
- other specifically binding molecules e.g. RNA molecules are used.
- Antibodies which recognize the protease-sensitive N-terminal region of the PrP are, for example, from "Brain Research, 545, (1991) 319-321 (antiserum anti-PrP-N),” Brain Pathol. 2002; 12: 1-11 (Antibodies FH11, BG4) "," Proc. Natl. Acad. Be. Vol. 95 pp. 8812-8815, July 1998 (Antikö ⁇ er 5B2) M or "Biochemical and Biophysical Research Communications 273, 136-139 (2000) (Antikö ⁇ er 8B4)".
- the publications mentioned describe the properties of the antibodies and their production.
- the complexes formed can be verified as standard. It is usually provided that one of the two components of the complex formed is carrier-bound.
- the sample material is immobilized after digestion, for example on a microtiter plate or on beads, and a detection with labeled molecules A and B, in particular antibodies A and B, is carried out.
- the preferably used antibodies can be incubated in one batch simultaneously or in succession with the sample material. It is equally possible to carry out two parallel runs, in each of which one of the two antibodies A or B is added.
- a sandwich immunoassay is preferably carried out for detection.
- two antibodies per analyte are used in such sandwich immunoassays, which are bound to two different epitopes of the analyte.
- One of the antibodies is usually immobilized and serves to couple the analyte to a solid phase, while the other serves as a detection antibody and is provided with a label.
- a further antibody C is also provided in this context, which recognizes PrP 27-30, although the epitope recognized by this antibody C is different than that recognized by Antikö ⁇ er B.
- Another possibility is to immobilize the antibodies A and B on a support and then, after incubation of the support with the sample material, to add labeled antibody C for detection.
- the sample material obtained after digestion is incubated in one batch first with immobilized antibodies A and then with immobilized antibodies B. Labeled antibody C is added for detection as described above.
- the staggering over time makes it possible in a simple manner that any protease-sensitive areas of the PrP that are present can initially bind to the specific antibodies A, without the binding kinetics being caused by a simultaneous attack by the antibodies serving as molecule B on the protease-resistant Area is disturbed.
- any complexes formed are detected using labeled molecules, in particular labeled antibodies. If a label can be detected or recognized on a support, this is a sign that the relevant antibody provided with the detected label has been bound, which, depending on the test setup, can be an indication of the presence of a certain complex.
- the molecules A and B or the antibody C can be e.g. the same or different fluorescent labels or with enzymes (Elisa) or with other suitable labels. Basically, all markings are suitable that can be directly or indirectly detected or measured.
- the person skilled in the art is aware of the different possibilities for labeling molecules, in particular antibodies, in a suitable manner for the above processes and for detecting them in the course of the processes. It should therefore not be dealt with in more detail at this point
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Hematology (AREA)
- Chemical & Material Sciences (AREA)
- Urology & Nephrology (AREA)
- Molecular Biology (AREA)
- Immunology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Neurosurgery (AREA)
- Neurology (AREA)
- Food Science & Technology (AREA)
- Cell Biology (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
Description
Claims
Priority Applications (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
NZ527233A NZ527233A (en) | 2001-04-21 | 2002-04-19 | Method for testing samples containing prion protein for the possible presence of PrPSc 27-30 |
US10/474,107 US20040115752A1 (en) | 2001-04-21 | 2002-04-19 | Method for testing samples containing prion protein for the possible presence of the prpsc form |
JP2002583988A JP2004528561A (ja) | 2001-04-21 | 2002-04-19 | プリオンタンパク質含有サンプルをPrPSc型の存在可能性について試験する方法 |
EP02737971A EP1381868A2 (de) | 2001-04-21 | 2002-04-19 | VERFAHREN ZUR UNTERSUCHUNG VON PRION-PROTEIN ENTHALTENDEN PROBEN AUF DAS EVENTUELLE VORLIEGEN DER PrP?Sc -FORM |
CA002437880A CA2437880A1 (en) | 2001-04-21 | 2002-04-19 | Method for testing samples containing prion protein for the possible presence of the prpsc form |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE10119713A DE10119713A1 (de) | 2001-04-21 | 2001-04-21 | Verfahren zur Untersuchung von Prion-Protein enthaltenden Proben auf das eventuelle Vorliegen der PrPSc-Form |
DE10119713.6 | 2001-04-21 |
Publications (2)
Publication Number | Publication Date |
---|---|
WO2002086511A2 true WO2002086511A2 (de) | 2002-10-31 |
WO2002086511A3 WO2002086511A3 (de) | 2003-07-24 |
Family
ID=7682316
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/EP2002/004341 WO2002086511A2 (de) | 2001-04-21 | 2002-04-19 | VERFAHREN ZUR UNTERSUCHUNG VON PRION-PROTEIN ENTHALTENDEN PROBEN AUF DAS EVENTUELLE VORLIEGEN DER PrPSc-FORM |
Country Status (7)
Country | Link |
---|---|
US (1) | US20040115752A1 (de) |
EP (1) | EP1381868A2 (de) |
JP (1) | JP2004528561A (de) |
CA (1) | CA2437880A1 (de) |
DE (1) | DE10119713A1 (de) |
NZ (1) | NZ527233A (de) |
WO (1) | WO2002086511A2 (de) |
Cited By (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
FR2849204A1 (fr) * | 2002-12-20 | 2004-06-25 | Afssa | Procede de detection de la prpsc utilisant un antibiotique d de la famille des aminoglycosides pour l'elimination et la detection de la prpsc dans des echantillons biologiques |
FR2849205A1 (fr) * | 2002-12-20 | 2004-06-25 | Afssa | Procede d'amplification de la detection de la prpsc et utilisation d'un ligand adjuvant macrocyclique pour une telle amplification |
EP1596199A1 (de) * | 2004-05-14 | 2005-11-16 | Prionics AG | Verfahren zum Nachweis von pathogenen Prionenproteinen. |
US7566530B2 (en) | 2004-01-20 | 2009-07-28 | Biomerieux | Method for detecting PrP using at least one positive charge and/or at least one glycosidic bond and a ligand other than a protein ligand |
WO2010084201A1 (en) * | 2009-01-26 | 2010-07-29 | Commissariat A L'energie Atomique Et Aux Energies Alternatives | Novel derivative of erythromycin for the treatment and diagnosis of prion disease |
US8158441B2 (en) | 2005-07-21 | 2012-04-17 | Biomerieux S.A. | Method for detecting aggregate-forming circulating protein forms using an agent for aggregating said forms and an agent for capturing formed aggregates |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE602005007458D1 (de) * | 2004-11-15 | 2008-07-24 | Roche Diagnostics Gmbh | Hoch-Durchsatz Prion Tests |
DE102007016324A1 (de) * | 2007-04-04 | 2008-10-09 | Priontype Gmbh & Co.Kg | Verfahren zum Nachweis von pathologisch verändertem Prion-Protein (PrPSc) |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1998035236A2 (en) * | 1997-02-06 | 1998-08-13 | Enfer Technology Limited | Immunological assay for spongiform encephalopathies |
FR2774988A1 (fr) * | 1998-02-16 | 1999-08-20 | Commissariat Energie Atomique | Procede de purification de la prpres a partir d'un echantillon biologique et ses applications |
WO2000029850A1 (en) * | 1998-11-17 | 2000-05-25 | Wallac Oy | An immunoassay for the determination of transmissible spongiform encephalopathies in bovine |
WO2001023425A1 (en) * | 1999-09-28 | 2001-04-05 | Universität Zürich | Factors having prion-binding activity in serum and plasma and agents to detect transmissible spongiform encephalopathitis |
-
2001
- 2001-04-21 DE DE10119713A patent/DE10119713A1/de not_active Withdrawn
-
2002
- 2002-04-19 WO PCT/EP2002/004341 patent/WO2002086511A2/de not_active Application Discontinuation
- 2002-04-19 CA CA002437880A patent/CA2437880A1/en not_active Abandoned
- 2002-04-19 US US10/474,107 patent/US20040115752A1/en not_active Abandoned
- 2002-04-19 JP JP2002583988A patent/JP2004528561A/ja active Pending
- 2002-04-19 EP EP02737971A patent/EP1381868A2/de not_active Withdrawn
- 2002-04-19 NZ NZ527233A patent/NZ527233A/en unknown
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1998035236A2 (en) * | 1997-02-06 | 1998-08-13 | Enfer Technology Limited | Immunological assay for spongiform encephalopathies |
FR2774988A1 (fr) * | 1998-02-16 | 1999-08-20 | Commissariat Energie Atomique | Procede de purification de la prpres a partir d'un echantillon biologique et ses applications |
WO2000029850A1 (en) * | 1998-11-17 | 2000-05-25 | Wallac Oy | An immunoassay for the determination of transmissible spongiform encephalopathies in bovine |
WO2001023425A1 (en) * | 1999-09-28 | 2001-04-05 | Universität Zürich | Factors having prion-binding activity in serum and plasma and agents to detect transmissible spongiform encephalopathitis |
Cited By (13)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7217530B2 (en) | 2002-12-20 | 2007-05-15 | Agence Francaise de Securite Sanitaire des Aliments-AFSSA | Process for detecting PrP using a macrocyclic adjuvant ligand |
FR2849205A1 (fr) * | 2002-12-20 | 2004-06-25 | Afssa | Procede d'amplification de la detection de la prpsc et utilisation d'un ligand adjuvant macrocyclique pour une telle amplification |
WO2004059321A1 (fr) * | 2002-12-20 | 2004-07-15 | Agence Francaise De Securite Sanitaire Des Aliments-Afssa- | Procede de detection de la prp utilisant un antibiotique des aminoglycosides |
WO2004059322A1 (fr) * | 2002-12-20 | 2004-07-15 | Agence Francaise De Securite Sanitaire Des Aliments-Afssa- | Procede de detection de la prp utilisant un ligand adjuvant macrocyclique. |
FR2849204A1 (fr) * | 2002-12-20 | 2004-06-25 | Afssa | Procede de detection de la prpsc utilisant un antibiotique d de la famille des aminoglycosides pour l'elimination et la detection de la prpsc dans des echantillons biologiques |
CN100380125C (zh) * | 2002-12-20 | 2008-04-09 | 法国食品卫生安全署 | 利用大环辅助配体检测prp的方法 |
US7695918B2 (en) | 2002-12-20 | 2010-04-13 | Agence Francaise de Securite Sanitaire des Aliments-AFSSA | Process for detecting PrPSC using an antibiotic from the family of aminoglycosides |
US7566530B2 (en) | 2004-01-20 | 2009-07-28 | Biomerieux | Method for detecting PrP using at least one positive charge and/or at least one glycosidic bond and a ligand other than a protein ligand |
EP2290093A2 (de) | 2004-01-20 | 2011-03-02 | Biomerieux | Verfahren zum Nachweis des Globalerwärmungspotenzials (GWP) mit Hilfe eines Moleküls, das mindestens eine positive Ladung hat und/oder mindestens eine Oxidverbindung und einen anderen Liganden als einen Proteinliganden |
EP1596199A1 (de) * | 2004-05-14 | 2005-11-16 | Prionics AG | Verfahren zum Nachweis von pathogenen Prionenproteinen. |
WO2005114214A1 (en) * | 2004-05-14 | 2005-12-01 | Prionics Ag | Method for the detection of disease-related prion |
US8158441B2 (en) | 2005-07-21 | 2012-04-17 | Biomerieux S.A. | Method for detecting aggregate-forming circulating protein forms using an agent for aggregating said forms and an agent for capturing formed aggregates |
WO2010084201A1 (en) * | 2009-01-26 | 2010-07-29 | Commissariat A L'energie Atomique Et Aux Energies Alternatives | Novel derivative of erythromycin for the treatment and diagnosis of prion disease |
Also Published As
Publication number | Publication date |
---|---|
JP2004528561A (ja) | 2004-09-16 |
NZ527233A (en) | 2005-07-29 |
DE10119713A1 (de) | 2002-10-24 |
WO2002086511A3 (de) | 2003-07-24 |
EP1381868A2 (de) | 2004-01-21 |
CA2437880A1 (en) | 2002-10-31 |
US20040115752A1 (en) | 2004-06-17 |
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