Method for detecting indicators for determining diseases

Patent metadata

Field	Value
Publication number	US-11175288-B2
Application number	US-201414900361-A
Country	US
Kind code	B2
Filing date	Jun 25, 2014
Priority date	Jun 26, 2013
Publication date	Nov 16, 2021
Grant date	Nov 16, 2021

Abstract

The invention concerns a method for detecting indicators for determining diseases (disease indicators), in which aggregates of misfolded proteins play a role, and a method for selective quantitation and/or characterization of these disease indicators.

First claim

What is claimed is:

1. A method for the qualitative and/or quantitative determination of disease indicators in which a sample is tested for at least two aggregate types of endogenous misfolded proteins, wherein the sample is tested for at least two different aggregate types on the same substrate without further processing and/or treatment and the method comprises:
(a) application of the sample to be tested to a substrate,
(b) addition of probes labeled for detection with fluorescent dyes, which probes label a respective aggregate by specifically binding to it, and
(c) detection of labeled aggregates by surface-based Fluorescence Intensity Distribution Analysis (sFIDA), carried out by a method with a temporally and spatially resolved signal and with high spatial resolution wherein a pixel is determined against a respective background, and wherein
(b) can be carried out before (a);
the disease is selected from tauopathies, AA (amyloid A) amyloidosis, AL (amyloid light chain) amyloidosis, AApoAI (amyloid apolipoprotein AI) amyloidosis, (amyloid apolipoprotein AII) AApoAII amyloidosis, ATTR (amyloid transthyretin) amyloidosis, schizophrenia and other DISC1 (disrupted in schizophrenia 1) opathies, amyotrophic lateral sclerosis, frontotemporal lobar degeneration and other FUS (fused in sarcoma) proteinopathies, diabetes mellitus type 2, Parkinson's disease and other synucleinopathies, chronic traumatic encephalopathy and other TDP-43 (transactive response DNA binding protein 43 kDa) proteinopathies, Huntington's disease, familial visceral amyloidosis and/or Alzheimer's dementia; and
the aggregate type of endogenous misfolded proteins is selected from tau aggregates, serum amyloid A protein aggregates, IgG (immunoglobulin G) light chain aggregates, AapoAI aggregates, AapoAll aggregates, ATTR aggregates, DISC1 aggregates, FUS aggregates, IAPP (islet amyloid polypeptide) aggregates, SOD1 (superoxide dismutase 1) aggregates, α-synuclein aggregates, TDP-43 aggregates, huntingtin aggregates, lysozyme aggregates, Aβ aggregates, and mixed aggregates.

2. The method of claim 1 , wherein prior to (a) capture molecules are immobilized on the substrate.

3. The method of claim 1 , wherein capture molecules immobilized on the substrate and/or the probes comprise specific antibodies to an epitope of the proteins which form the aggregates.

4. The method of claim 1 , wherein the sample is tested for at least three aggregate types of endogenous misfolded proteins.

5. The method of claim 1 , wherein the method is a method for a differential diagnosis of a disease selected from tauopathies, AA amyloidosis, AL amyloidosis, AApoAI amyloidosis, AApoAII amyloidosis, ATTR amyloidosis, schizophrenia and other DISC1opathies, amyotrophic lateral sclerosis, frontotemporal lobar degeneration and other FUS proteinopathies, diabetes mellitus type 2, Parkinson's disease and other synucleinopathies, chronic traumatic encephalopathy and other TDP-43 proteinopathies, Huntington's disease, familial visceral amyloidosis and/or Alzheimer's dementia vs. another disease selected from the above-mentioned diseases and comprises:
(i) quantitative determination of disease indicators according to claim 1 ;
(ii) comparison of obtained data with standard values;
(iii) detection of a significantly different quantity of disease indicators as discrepancy in the comparison; and
(iv) attribution of the discrepancy to a disease selected from the above-mentioned diseases.

6. The method of claim 1 , wherein the method is a method for a differential diagnosis and wherein after a quantification of a disease indicator for a disease selected from tauopathies, AA amyloidosis, AL amyloidosis, AApoAI amyloidosis, AApoAll amyloidosis, ATTR amyloidosis, schizophrenia and other DISC1opathies, amyotrophic lateral sclerosis, frontotemporal lobar degeneration and other FUS proteinopathies, diabetes mellitus type 2, Parkinson's disease and other synucleinopathies, chronic traumatic encephalopathy and other TDP-43 proteinopathies, Huntington's disease, familial visceral amyloidosis and/or Alzheimer's dementia, obtained data are compared with standard values, a significantly different quantity of disease indicators is detected as discrepancy in the comparison and the discrepancy is attributed to an above-mentioned disease.

7. The method of claim 1 , wherein the aggregates comprise small, freely diffusing oligomers.

8. The method of claim 1 , wherein a quantitative determination of disease indicators is carried out, which determination comprises a determination of composition, size and/or shape of aggregates.

9. The method of claim 7 , wherein a quantitative determination of disease indicators is carried out, which determination comprises a determination of composition, size and/or shape of aggregates.

10. The method of claim 1 , wherein the aggregates comprise one or more peptides or monomers of SEQ ID Nos: 1-9 and 12-14.

Assignees

• Forschungszentrum Juelich Gmbh

Inventors

• Willbold Dieter
• Funke Susanne Aileen
• Birkmann Eva
• Kravchenko Kateryna
• Bannach Oliver
• Korth Carsten
• Bader Verian
• Huebinger Steffen

Classifications

• G01N33/6896 Primary

  Neurological disorders, e.g. Alzheimer's disease · CPC title

• G01N21/6456

  Spatial resolved fluorescence measurements; Imaging · CPC title

• G01N2333/936

  acting on beta-1, 4 bonds between N-acetylmuramic acid and 2-acetyl-amino 2-deoxy-D-glucose, e.g. lysozyme · CPC title

• G01N2800/302

  Schizophrenia · CPC title

• G01N2333/4709

  Amyloid plaque core protein · CPC title