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Methods for analyzing blood to detect diseases associated with abnormal protein aggregation

A protein aggregation and protein technology, applied in proteomics, analytical materials, disease diagnosis, etc., can solve problems such as invasive, expensive, and unable to withstand high-throughput analysis

Active Publication Date: 2016-01-13
彼德·史戴斯 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This has not been proven to be reliable because the levels of Aβ40, Aβ42 or the ratio of the two cannot reliably distinguish between healthy controls and AD patients (Thambisety and Lovestone 2010)
Other approaches include proteomic analysis of plasma, but it is expensive, complex, not amenable to high-throughput analysis and remains experimental
CSF analysis of Aβ and (phosphorylated) microscopically associated (tau) levels have stronger predictive value (vanRossum et al., 2012; Senanaron et al., 2012), but are invasive and less likely than blood Becoming a routine method outside of formal research trials

Method used

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  • Methods for analyzing blood to detect diseases associated with abnormal protein aggregation
  • Methods for analyzing blood to detect diseases associated with abnormal protein aggregation
  • Methods for analyzing blood to detect diseases associated with abnormal protein aggregation

Examples

Experimental program
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Effect test

Embodiment 1

[0140] Example 1. Detection of Alzheimer's disease from blood using fluorescence spectroscopy

[0141] Many neurological diseases, such as AD, Parkinson's disease, Huntington's disease and prion diseases, are associated with the misfolding and aggregation of proteins from their soluble functional state into highly ordered fibrous assemblies with high β-sheet content. These pathologically unrelated diseases have in common a specific binding to the classic amyloid probe, Congo red, and are therefore termed "Congo red-ophilic" diseases. Conformational-sensitive fluorescent probes that specifically bind amyloid fibrils have been widely used to investigate protein aggregation (Styren et al., 2000; Klunk et al., 2002; Mathis et al., 2002; Aslund et al., 2009; Nilsson et al., 2005; Hammarstrom et al., 2010). Some newer probes have the additional property of detecting soluble oligomer species and altering their emission spectra based on the protein aggregates to which they bind, and ...

Embodiment 2

[0148] Example 2: Detection of protein aggregates in leukocytes using probe K114

[0149] Protein aggregates were detected in human leukocytes using the fluorescent probe K114, an amyloid-specific dye and an analog of Congo red. In addition, the fluorescent signal from labeled leukocytes is processed using an algorithm (e.g., an algorithm based on the Levenberg-Marquardt damped least squares nonlinear curve fitting algorithm) to separate the disease-specific signal from background and autofluorescence emission . Fluorescent emission-based counts are calculated, reflecting the likelihood that the test cell sample originated from a patient with a disease associated with abnormal protein aggregation.

[0150] Program

[0151] Blood was drawn from the patient into EDTA tubes and kept on ice. The buffy coat (white blood cells) was isolated using standard techniques. Briefly, the blood samples were centrifuged at 1700 RCF at room temperature to remove a band of concentrated leuk...

Embodiment 3

[0163] Example 3. Presence of amyloid in the brains of MS patients

[0164] AD is generally considered to be a neurodegenerative disorder with secondary innate inflammation, but the perspective on multiple sclerosis is less clear. Initially, the evidence strongly supported a primary autoimmune disorder, the resulting inflammatory attack triggering demyelination and cortical atrophy. However, recent data from detailed pathological examination and experience with effective anti-inflammatory treatments cast doubt on this conclusion (Stys et al., 2012).

[0165] Thus, the possibility exists that MS may also be a primary degenerative disorder, with an unusually prominent inflammatory secondary response. So what could be the root cause of MS? Whereas the pathology of later chronic MS (in which the inflammatory response is less prominent) shows the development of demyelinating lesions over time (possibly spreading from discrete foci in plaques?), the pathology of the periventricula...

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Abstract

A method of detecting a disease associated with abnormal protein aggregation in a subject is provided, the method comprising (a) contacting leukocytes from the subject with a probe that binds to pathogenic protein aggregates, and (b) detecting the probe bound to the pathogenic protein aggregates, wherein the presence of pathogenic protein aggregates in the leukocytes is indicative that the subject has a disease associated with abnormal protein aggregation. In one embodiment, the disease is Alzheimer's disease, mild cognitive impairment or traumatic brain injury.

Description

[0001] related application [0002] This application claims priority to US Patent Application No. 13 / 833008, filed March 15, 2013, the contents of which are hereby incorporated by reference in their entirety. technical field [0003] The present disclosure relates to methods for analyzing blood to detect diseases associated with abnormal protein aggregation. In one embodiment, the disease associated with abnormal protein aggregation is Alzheimer's disease. Background technique [0004] Alzheimer's disease (AD) is the most common cause of dementia in the elderly population, affecting more than 35 million people worldwide and is expected to rise to 115 million by 2050 unless effective treatments are developed (Barnes and Yaffe 2011). This age-related neurodegenerative disorder is pathologically characterized by amyloid beta (Aβ), including senile plaques, neurofibrillary tangles and synaptic loss in the brain (Selkoe 2001). While it is clear that AD is a degenerative disor...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N21/64G01N33/52G01N33/68G16B20/00
CPCG01N2800/2814G01N2800/2821G01N2800/2828G01N2800/2835G01N2800/2842G01N2800/285G16B20/00G01N33/6896G01N2333/4709
Inventor 彼德·史戴斯筒井茂树
Owner 彼德·史戴斯
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