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Protein markers of coronary heart disease in urine and its use

A technology for coronary heart disease and its application, which is applied in the field of protein markers of coronary heart disease in urine, and can solve the problems of abnormal expression level, unclear mechanism of action, and no clear report on the correlation of coronary heart disease

Active Publication Date: 2019-10-25
THE INST OF BASIC MEDICAL SCI OF CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition to the identification of coronary heart disease-related proteins such as APOE, it was also found that although other proteins in the homologous family have been reported to be associated with cardiovascular and cerebrovascular diseases, there is no clear report on the correlation between TFF1 and APOD in urine and coronary heart disease
Among them, TFF1 (clover factor 1) family protein sequence is characterized by a 40-amino acid fragment containing three conserved disulfide bonds, and is mainly distributed in the epithelium of the digestive system. Its specific mechanism of action is not yet clear, but it is found in The expression level of this protein is abnormal in a variety of tumors, and the expression of this protein in coronary heart disease has not been reported

Method used

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  • Protein markers of coronary heart disease in urine and its use
  • Protein markers of coronary heart disease in urine and its use
  • Protein markers of coronary heart disease in urine and its use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 Screening of Coronary Heart Disease Related Proteins in Urine

[0040] Urine was screened for coronary heart disease-related proteins using isotope-labeled relative and absolute quantification (iTRAQ) labeling differential proteomics.

[0041] Materials and Reagents

[0042] 1) Instrument: ultra-high performance liquid chromatography (Waters Company), model ACQUITY UPLC H-Class. TripleTOF mass spectrometer (AB sciex company), model 5600.

[0043] 2) Main reagents: Pierce BCA Protein Quantitative Analysis Kit (Thermo Scientific); trypsin (Promega); C18 solid-phase extraction cartridge (3CC, 60 mg, Waters); 4 standard iTRAQ reagents (AB sciex); C18 reverse-phase chromatographic column (4.6mm × 250mm, C18, 3μm, Waters Company).

[0044] 3) Sample: obtained from Peking Union Medical College Hospital.

[0045] 1.1 Preparation of urine protein

[0046] Urine samples were centrifuged at 5000 g for 30 min at 4 °C to remove cell debris. Take the supernatant, add ...

Embodiment 2

[0053] Example 2 Mass Spectrometry Detection of Urinary Protein TFF1 and APOD

[0054] In order to better carry out the application detection of these differential proteomic analysis results in further larger-scale clinical samples, the inventors used the target mass spectrometry analysis method to monitor the differential proteins identified. Multiple reaction monitoring (Multiple Reaction Monitor, MRM) is a target mass spectrometry quantitative analysis technology based on the signal of secondary mass spectrometry. It does not require special chemical labeling assistance, and can simultaneously perform absolute and relative quantitative determination of multiple proteins.

[0055] Materials and Reagents

[0056] 1) Instrument: ultra-high performance liquid chromatography (Waters Company), model ACQUITY UPLC H-Class. TripleTOF mass spectrometer (AB sciex company), model 5600.

[0057] 2) Main reagents: Pierce BCA Protein Quantitative Analysis Kit (Thermo Scientific); trypsi...

Embodiment 3

[0077] Example 3 Analysis of mass spectrometry detection results based on urine protein TFF1 and APOD combined with other urine proteins

[0078] In addition to urine TFF1 and APOD, 17 other proteins were also detected by mass spectrometry at the same time. Taking AMPE as an example, ROC curve analysis was performed on the protein combination, and the results showed the AUC analysis results of the combination of TFF1, APOD and AMPE. was 0.95, its specificity was 0.92, and its sensitivity was 0.82. Compared with only using TFF1 and APOD, there was no significant improvement, but this operation showed that integrated analysis could be performed based on the combination of TFF1 and APOD with other urine proteins. For specific ROC curves, see Figure 5 .

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Abstract

The invention discloses a coronary heart disease protein marker in urine and an application of the marker, and further discloses an application of an identification reagent selected from proteins suchas TFF1, APOD and combinations thereof to preparation of a reagent for diagnosing coronary heart diseases of subjects. The two proteins have good application prospects in large-scale screening of thecoronary heart disease, and have the advantage that materials are drawn non-invasively by the aid of the urine.

Description

technical field [0001] The present invention relates to the field of biotechnology, more specifically, to protein markers of coronary heart disease in urine and uses thereof. Background technique [0002] Coronary atherosclerotic heart disease is referred to as coronary heart disease for short. Coronary heart disease characterized by atherosclerosis and its complications have a high morbidity and mortality rate, which has attracted widespread attention. Accompanied by the course of the disease, atherosclerosis will develop into myocardial infarction and stroke in a certain period of time, directly threatening people's lives. Early diagnosis of the disease before it gets worse is crucial for the control and treatment of the disease. At present, early diagnosis methods of cardiovascular diseases, such as electrocardiography, ultrasound, traditional coronary angiography, coronary computed tomography angiography, etc., have good sensitivity and specificity for monitoring the se...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68
CPCG01N33/6848G01N33/6893G01N2800/324
Inventor 孙伟曾勇孙海丹王丹琪刘东方郭正光邵晨
Owner THE INST OF BASIC MEDICAL SCI OF CHINESE ACAD OF MEDICAL SCI