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Biochemical markers for CVD risk assessment

A use, antibody technology, applied in the field of biochemical markers for CVD risk assessment

Active Publication Date: 2018-04-10
NORDIC BIOSCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in some cases, a more invasive approach, i.e. angioplasty or bypass surgery, may be required

Method used

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  • Biochemical markers for CVD risk assessment
  • Biochemical markers for CVD risk assessment
  • Biochemical markers for CVD risk assessment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0265] To analyze the localization of proteoglycans and proteases, we performed immunohistochemical staining on human arterial samples from the left anterior descending coronary artery (LAD). In the following, it is demonstrated that the cathepsin K protease colocalizes with biglycan.

[0266] Such as figure 1 and 2 Immunohistochemical staining shown in shows colocalization of biglycan with cathepsin K. This may indicate that biglycan is a preferred substrate for cathepsin K. The same immunohistochemical staining was performed on samples of aorta in which atherosclerotic plaques formed and consequently resulted in macrophage foam cell infiltrates and calcifications replacing normal aortic structures. The results of these immunostainings are summarized in image 3 and 4 middle.

[0267] Immunohistochemical staining of biglycan and cathepsin K revealed co-localization in developing atherosclerotic lesions. Combining these results led to the hypothesis that a specific cle...

Embodiment 2

[0269] Biglycan degradation for assessment of degraded fragments. Biglycan from bovine articular cartilage (B8041-Sigma-Aldrich) was cleaved by the following proteases: MMP2, MMP3, MMP8, Cathepsin K, Cathepsin S, Cathepsin B, and Cathepsin L. The proteoglycan fragments produced by the enzymatic cleavage of the above proteases were treated at 10% Separation on a Bis-Tris gel followed by silver staining by the "Silver Express" silver staining kit (Invitrogen Cat# LC6100, Lot# 341099). Separation of proteolytically produced biglycan and silver staining results in Figure 5 displayed in .

Embodiment 3

[0271] Mice were immunized with peptides from type III collagen conjugated to ovalbumin. Serum was screened for reactivity with biotin-conjugated screened peptide sequences. Monoclonal antibody-secreting clones are generated and screened using the selection sequence. Clones were examined for lack of reactivity with an extended form of the target peptide that continues an adjacent sequence in type III collagen (reverse selection peptide) and for lack of reactivity with a nonsense peptide. All target sequence positive clones did not react with extended or nonsense sequences.

[0272] The target sequence, immunogen, screening sequence and counter-selection sequence are as follows:

[0273]

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Abstract

The present invention relates to biochemical markers for CVD risk assessment and bioassays for the quantification of peptide fragments containing neo-epitopes that are cleaved by proteases from proteins of atherosclerotic plaques For example, leucin, versican, perlecan, decorin, biglycan, collagen type III, CRP, ApoE or elastin, the method comprises the sample For example urine or serum is contacted with antibodies reactive to the neo-epitope and the level of binding of the immunological binding partner to the peptide fragment in the sample is determined. The assay predicts the risk of a cardiovascular disease event.

Description

[0001] This application is a divisional application of Chinese patent application CN200880123912.0, and the original application is an application that entered the Chinese national phase on July 2, 2010 with the international application number PCT / EP2008 / 064946. technical field [0002] The present invention relates to assays for the detection of biochemical markers valuable for diagnostic purposes of cardiovascular disease and prognosis of disease progression, including biochemical markers indicative of the risk of cardiovascular events resulting from the development of atherosclerosis and plaque instability thing. Background technique [0003] Cardiovascular disease (CVD) is the leading cause of disease and death worldwide. Currently, there is no efficient and non-invasive diagnostic method that allows diagnosis and classification of patients into different risk groups, as well as allowing diagnosis of low-risk patients. Diagnostic and prognostic tools primarily consist ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68
CPCG01N33/68C07K14/775G01N33/542G01N2800/32C12Q1/37C07K14/77C07K14/78G01N33/577G01N33/6878G01N33/6893G01N2800/50G01N2800/52
Inventor 莫滕·卡尔斯达尔佩尔·奎斯特娜塔莎·巴拉斯丘克
Owner NORDIC BIOSCI
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