Characteristic protein maker composition for mass spectroscopic diagnosis of thalassemia and diagnostic products thereof
A thalassemia and characteristic protein technology, which is applied in the field of characteristic hemoglobin composition or mass spectrometry model, and can solve problems such as inability to realize globin chain monitoring and the like
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Embodiment 1
[0099] Example 1. Mass Spectrometry Detection of Thalassemia Whole Blood Polypeptides and Establishment of Thalassemia Screening Model
[0100] 1. Samples and instruments:
[0101] Selected from 300 whole blood samples, including 150 cases from thalassemia patients, another 150 cases from healthy people, and 10 newborn umbilical cord blood samples. All patients with thalassemia were confirmed by postoperative pathology report. All whole blood samples were drawn in the morning on an empty stomach (umbilical cord blood for newborns), and the whole blood was separated and stored in a -80°C freezer.
[0102] Matrix-assisted laser desorption time-of-flight mass spectrometry Clin-TOF was developed by Beijing Yixin Bochuang Biotechnology Co., Ltd. Use the DP data analysis software developed by the company to process the data.
[0103] 2. Technical route:
[0104] Whole blood collection: Collect blood samples in BD tubes to avoid hemolysis. Slowly shake the tube up and down five ...
Embodiment 2
[0130] Embodiment two, the identification of characteristic protein
[0131] Magnetic bead enrichment method
[0132] 1. After the peak to be identified is determined according to the analysis, back check the sample with the highest intensity of the peak to be identified among the pre-processed samples.
[0133] 2. Identify the magnetic beads used in previous experiments. Treat 20 copies of this sample in parallel. Enrichment into one tube.
[0134] 3. Centrifuge at 1300rmp for 5min. Take the supernatant on a magnetic rack. Avoid leaving magnetic beads to affect later experiments.
[0135] 4. Spin the liquid dry and mark.
[0136] Adopt the Nano Aquity UPLC liquid phase system of Waters Company: the parameters are set as follows, trapping column: C18, 5μm, 180μm×20mm, nanoAcquity TM Column; analysis column: C 18 , 1.7μm, 75μm×150mm, nanoAcquity TM Column; mobile phase A: 5% acetonitrile, 0.1% formic acid in water; mobile phase B: 95% acetonitrile, 0.1% formic acid...
Embodiment 3
[0153] Embodiment 3, the blind selection test of thalassemia screening model
[0154] 200 of 300 whole blood samples (150 from thalassemia patients and 150 from healthy people) were selected as training samples for model building. All patients with thalassemia were identified by genetic testing. All whole blood samples were drawn in the morning on an empty stomach, separated and stored in a -80°C freezer.
[0155] Another 100 remaining samples were selected as test samples for blind selection test, 50 of which were known to be from thalassemia patients and the other 50 were from healthy people. The processing method is the same as above.
[0156] The mass spectrometry model of the thalassemia polypeptide was established by using the characteristic polypeptide peaks of thalassemia screened in Examples 1-2. The model is determined to use 3 input variables, namely: 15120m / z, 15859m / z, 16952m / z.
[0157] The recognition rate of model training is 100%. And the random selection...
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