Protein marker for myocardial infarction in urine, and applications thereof in diagnosis and prognosis
A myocardial infarction, protein technology, applied in biological tests, instruments, measurement devices, etc., can solve the problem of not being able to detect the disease process and treatment effect in time
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Embodiment 1
[0056] Example 1: Detection of myocardial infarction-related proteins in urine
[0057] We used isotope labeling relative and absolute quantification (iTRAQ) labeling differential proteomics to screen for myocardial infarction-related proteins in urine.
[0058] Materials and Reagents
[0059] 1) Instrument: Triple TOF 5600 mass spectrometer (Absciex Company).
[0060] 2) Main reagents: trypsin (Promega); C18 solid-phase extraction column (3CC, 60mg, Waters); 4 standard iTRAQ reagents (Absciex); C18 reversed-phase chromatography column (4.6mm×250mm, C18, 3μm , Waters Corporation).
[0061] 3) Samples: urine from 7 patients with myocardial infarction (within 12 hours of onset and 7 days after treatment) and 7 normal controls, from the Fourth Affiliated Hospital of Jilin University.
[0062] 1.1 Preparation of urine protein
[0063] Urine samples were centrifuged at 3,500 g for 15 min at 4°C to remove cell debris. Take the supernatant, add pre-cooled absolute ethanol (-20°C...
Embodiment 2
[0070] Example 2: Mass Spectrometric Detection of Urinary Protein Antithrombin-III, Complement C3, α-1-Acid Glycoprotein 1, Serum Transferrin and Cathepsin Z
[0071] In order to better carry out the application detection of these differential proteomic analysis results in further large-scale clinical samples, we used targeted mass spectrometry to monitor the differential proteins identified. Multiple Reaction Monitor (MRM) is a target mass spectrometry quantitative analysis technology based on MS / MS signals, which can simultaneously perform absolute and relative quantitative determinations without the assistance of special chemical labels.
[0072] Materials and Reagents
[0073] 1) Instrument: QTRAP 6500 mass spectrometer (AB Sciex Company).
[0074] 2) Main reagents: trypsin (Promega Company); C18 solid-phase extraction cartridge (3CC, 60 mg, Waters Company).
[0075] 3) Samples: urine from 15 patients with myocardial infarction (within 12 hours of onset and seven days af...
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