Diagnostic kit for diagnosing disorders or diseases related to abnormal protein aggregation or misfolding of protein using dissolution of protein aggregates

A diagnostic kit, misfolding technology, used in peptide/protein components, biological testing, disease diagnosis, etc., can solve problems such as high cost, difficulty in accurately diagnosing Alzheimer's disease, and invasive costs

Inactive Publication Date: 2016-02-17
KOREA INST OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although symptoms can be checked indirectly based on physical, neurological, or physiological tests, measurements of β-amyloid in cerebrospinal fluid, and most accurately by brain imaging to monitor structural and functional changes in the brain and to identify plaques of β-amyloid to diagnose the progression of Alzheimer's disease, but these methods are very invasive and expensive
Due to these disadvantages, it is difficult to accurately diagnose Alzheimer's disease

Method used

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  • Diagnostic kit for diagnosing disorders or diseases related to abnormal protein aggregation or misfolding of protein using dissolution of protein aggregates
  • Diagnostic kit for diagnosing disorders or diseases related to abnormal protein aggregation or misfolding of protein using dissolution of protein aggregates
  • Diagnostic kit for diagnosing disorders or diseases related to abnormal protein aggregation or misfolding of protein using dissolution of protein aggregates

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0104] Diagnostic kit for measuring the concentration of aggregated protein before and after dissociation

[0105] 1. Preparation of Mice

[0106] Female APP / PS1 transgenic mice were used as test animals, and a total of 32 mice (11 at 5-6 months and 21 at 7.2-8.6 and 13 months) were used. The use of 5-6 month old mice was due to the accumulation of β-amyloid plaques in the brain starting from 5 months.

[0107] The transgenic APP / PS1 mouse model was B6C3-Tg (APPswe, PSEN1dE9) 85Dbo / Mmjax mice purchased from Jackson Laboratory (USA), which were bred and used in experiments according to the instructions of the Experimental Animal Ethics Committee.

[0108] EPPS was administered to each of 31 mice at a dose of 1,000 mg / kg / day.

[0109] 2. Blood draw and measurement of β-amyloid concentration

[0110] Before and after EPPS administration, blood was drawn from the retro-orbital vein of the mice using microtubes (Marienfeld, Germany) treated with 80 IU / mL heparin. Blood d...

Embodiment 2

[0127] Diagnostic kit for measuring dissociated concentration of aggregated protein in blood

[0128] Measurement of β-amyloid monomer concentration and diagnosis of Alzheimer's disease using APP / PS1 / Tau transgenic mice

[0129] 1. Preparation of Mice

[0130] 5-month-old female APP / PS1 / Tau transgenic mice were used as test animals.

[0131] The transgenic APP / PS1 / Tau mouse model was B6;129-Psen1tm1MpmTg(APPSwe, tauP301L)1Lfa / Mmjax mice purchased from Jackson Laboratory (USA), which were bred and used in experiments according to the guidance of the Experimental Animal Ethics Committee.

[0132] 2. Plasma and Whole Blood Processing

[0133] Blood was collected in K2EDTA-treated vacuum tubes containing Roche Complete Mini (protease inhibitor) solution and centrifuged. Plasma separated from the cell fraction was collected in Eppendorf tubes and made into 0.1 mL samples for immediate use or stored at −80 °C for later use. Whole blood was collected in Eppendorf tubes, tr...

Embodiment 3

[0146] Diagnostic kit for measuring dissociated concentration of aggregated protein in blood

[0147] Measurement of β-amyloid oligomer concentration using APP / PS1 transgenic mice

[0148] 1. Preparation of Mice

[0149] Nine-month-old female APP / PS1 transgenic mice were used as test animals.

[0150] The transgenic APP / PS1 mouse model was B6C3-Tg(APPswe, PSEN1dE9)85Dbo / Mmjax mice purchased from Jackson Laboratory (USA), which were bred and used in experiments according to the guidance of the Experimental Animal Ethics Committee.

[0151] Sample processing and concentration measurement of β-amyloid were performed in the same manner as in Example 1.

[0152] Table 7 shows the monomer concentration in the plasma of the control group (UPAβ), the monomer concentration in the plasma of the EPPS group (MPAβ) and the monomer concentration in the whole blood of the EPPS group (MBAβ), as well as the UP / MP and MB / MP values . The average values ​​of the three measurements are sh...

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Abstract

The present invention relates to a diagnostic kit for diagnosing disorders or diseases related to abnormal protein aggregation or misfolding of protein by using dissolution of protein aggregates in the body, capable of accurately diagnosing disorders or diseases such as Alzheimer's disease caused by aggregation of beta amyloid, as well as disorders and diseases related to abnormal protein aggregation or misfolding of protein by analyzing concentrations through the dissolution of aggregated protein in the blood of a person having other disorders or diseases due to protein aggregation.

Description

technical field [0001] The present disclosure relates to a blood diagnostic kit capable of accurately diagnosing diseases or conditions associated with abnormal aggregation or misfolding of proteins, including those caused by aggregation of β-amyloid protein, based on the concentration analysis of aggregated proteins before and after dissociation conditions or diseases such as Alzheimer's disease and conditions or diseases caused by aggregation of other proteins. Background technique [0002] Neuronal dysfunction and damage can be induced by toxic and aggregation-prone proteins, which can manifest in many neurological disorders. Such conditions include, for example, amyotrophic lateral sclerosis, Alzheimer's disease, Parkinson's disease, prion diseases, polyglutamine expansion disease, spinocerebellar ataxia, spinal and bulbar muscular atrophy, spongiform encephalopathy , tauopathy, Huntington's disease, or dystonia. [0003] The toxic and aggregation-prone proteins that c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/53
CPCG01N33/6896G01N2333/4709G01N2800/2821A61K38/1716C12Q1/6886G01N1/28C07K14/4711G01N33/6827G01N33/6893
Inventor 金泳秀金东辰金惠渊曹秀民金泰松金贤真李世真
Owner KOREA INST OF SCI & TECH
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