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Biomarker of Parkinson disease

A technology for biomarkers and Parkinson's disease, applied in the direction of biological testing, microbiological determination/examination, biochemical equipment and methods, etc., can solve problems such as complex diagnosis, difficult diagnosis of Parkinson's disease, lack of clinical symptoms, etc.

Inactive Publication Date: 2010-03-03
生物远景技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in its early stages, Parkinson's disease is difficult to diagnose due to lack of clinical symptoms and lack of reliable biomarkers
In addition, Parkinson's disease can closely resemble other disorders, including but not limited to, such as essential tremor and multiple system atrophy, complicating the possible diagnosis

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0109] Example 1 - Separation of proteins and identification of protein variants / isoforms using two-dimensional gel electrophoresis

[0110] Serum protein samples were analyzed by two-dimensional gel electrophoresis. Separation in the first direction was performed by adding 1 mg of protein sample to 24 cm IPG gel strips using a non-linear pH gradient of 3-7. Protein samples were separated using an IPGphor device (GE Healthcare, Piscataway, NJ) at 20°C by pressurizing to 130,000Vh under the following conditions: 40V for 10 hours; 100V for 20 hours; 200V for 2 hours; 500V for 2 hours; 1,000V 2 hours; 8,000V gradient for 2 hours; 8,000V for 12.5 hours. Then the IPG strips were incubated in equilibration buffer (50mM Tris-HCl, 6M urea, 30% (volume / volume) glycerol, 2% (weight / volume) SDS, trace bromophenol blue, 1% (weight / volume) di Thiothreitol) was incubated for 15 minutes. The IPG strips were incubated for an additional 15 minutes in equilibration buffer containing 2.5% (...

Embodiment 2-W

[0114] Example 2 - Western blot validation

[0115] To verify the results obtained in Example 1, sera from PD patients and controls were further subjected to Western blot analysis. Serum protein samples (50 micrograms) were separated by 12.5% ​​SDS-PAGE and transferred to PVDF membranes (Bio-Rad, Hercules, CA). Membranes were blocked and incubated overnight at 4°C with TTR polyclonal antibody (1:5000, Santa Cruz Biotechnology, Santa Cruz, CA). Anti-rabbit IgG-HRP secondary antibody (Santa Cruz Biotechnology, Santa Cruz, CA) was added at a ratio of 1:10,000. The relative intensities of the bands were calculated with QuantityOne software (Bio-Rad, Hercules, CA, version 4.6.3). Such as Figure 3A As indicated, the presence of TTR was detected in serum proteins (50 mg) of PD patients (lanes 1-3) or controls (lanes 4-6). As a positive control, 50 ng of purified human TTR was added to a separate lane (+). Molecular weight markers are indicated with corresponding KDa masses. ...

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Abstract

The invention relates to a biomarker of Parkinson disease, in particular to a method used for detecting pathogeny of the Parkinson disease and identifying the biomarker of the Parkinson disease (PD).The invention also provides a kit used for diagnosing or monitoring PD. A proteinic isoform or a monomer or polymer form thereof can be used for early diagnosing vulnerable subjects who are possible to suffer from PD. For example, the increase of the level of monomer transthyretin (TTR) in serum or the reduction of the level of dipolymer TTR can be used as a tool for diagnosing and monitoring PD.

Description

technical field [0001] The present invention relates generally to the fields of molecular biology, neurobiology, pharmacology and diagnosis, prognosis or prevention / treatment of Parkinson's disease (PD), and in particular to biomarkers for Parkinson's disease. Background technique [0002] Parkinson's disease is a progressive neurodegenerative disorder characterized by the loss of dopaminergic neurons in the substantia nigra and the presence of inclusion bodies (Lewy bodies) in surviving neurons. Typical clinical features of Parkinson's disease include progressive tremor, rigidity, and bradykinesia. However, in its early stages, Parkinson's disease is difficult to diagnose due to lack of clinical symptoms and lack of reliable biomarkers. In addition, Parkinson's disease can closely resemble other disorders, including but not limited to, such as essential tremor and multiple system atrophy, complicating the possible diagnosis. A diagnosis of PD can be made on the basis of n...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/48C12Q1/00G01N33/53G01N27/447G01N27/64
Inventor 陈彪
Owner 生物远景技术有限公司
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