Method for producing a standard for detecting protein aggregates of a protein misfolding disease, standard and use thereof

Patent metadata

Field	Value
Publication number	US-10948499-B2
Application number	US-201615548266-A
Country	US
Kind code	B2
Filing date	Feb 25, 2016
Priority date	Mar 18, 2015
Publication date	Mar 16, 2021
Grant date	Mar 16, 2021

Abstract

An inorganic nanoparticle is provided (step A) having the size of the aggregate of the protein misfolding disease. Free amino groups or free carboxyl groups are formed (step B) on the surface of the nanoparticle (for functionalizing the nanoparticle surface into an amine- or carboxy-functionalized nanoparticle. Maleinimido spacer carboxylic acid is bound to the free amino groups in step (B). Or, free carboxyl groups in step (B) are converted into NHS esters. Monomers of the protein aggregate are bound i) to the maleinimido spacer carboxylic acids by way of a sulfhydryl group at the free end of the monomers, or ii) to the NHS esters by way of the amino group at the free end of the monomer. A standard is provided for use in the detection of protein aggregates occurring with protein misfolding diseases.

First claim

The invention claimed is:

1. A method of using a standard for the detection of a protein aggregate of Alzheimer's disease, comprising:
providing an inorganic silica nanoparticle having the size 15 to 200 nm;
forming free amino groups on the surface of the inorganic silica nanoparticle for functionalizing the inorganic silica nanoparticle surface into an amine-functionalized nanoparticle;
binding a maleinimido spacer carboxylic acid to the free amino groups;
binding an amyloid beta monomer of the protein aggregate to the maleinimido spacer carboxylic acid by way of a sulfhydryl group at a free end of the amyloid beta monomer of the protein aggregate, thereby producing the standard comprising a known number of binding sites;
providing a sample comprising an unknown number of binding sites;
treating the sample and the standard with a fluorescent-labeled antibody; and
inferring the number of binding sites in the sample from the standard treated with said fluorescent-labeled antibody.

Assignees

• Forschungszentrum Juelich Gmbh

Inventors

• HUELSEMANN Maren
• Zafiu Christian
• Bannach Oliver
• Willbold Dieter

Classifications

• G01N33/96

  involving blood or serum control standard · CPC title

• G01N33/54346 Primary

  Nanoparticles · CPC title

• G01N33/6896 Primary

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

• G01N2800/2821

  Alzheimer · CPC title

• G01N2333/4709

  Amyloid plaque core protein · CPC title