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Application of characteristic protein composition or mass spectrum model for representing thalassemia

A technology for thalassemia and characteristic proteins, which is applied in the field of characteristic hemoglobin compositions or mass spectrometry models, and can solve problems such as the inability to monitor globin chains

Inactive Publication Date: 2020-01-07
长沙湘华质谱医学科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this method uses MALDI-TOF-MS detection, its purpose is to detect the combined products of hemoglobin and blood sugar, and to conduct accurate quantitative analysis through the difference in the mass-to-charge ratio of glycosylated hemoglobin and non-glycosylated hemoglobin. Monitoring of conformation of globin chains

Method used

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  • Application of characteristic protein composition or mass spectrum model for representing thalassemia
  • Application of characteristic protein composition or mass spectrum model for representing thalassemia
  • Application of characteristic protein composition or mass spectrum model for representing thalassemia

Examples

Experimental program
Comparison scheme
Effect test

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 ac...

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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Abstract

The invention provides a mass spectrum model for detecting a characteristic protein fragment composition of thalassemia and evaluating a therapeutic effect of a thalassemia drug or evaluating an effect of a treatment method. A sequence of the characteristic protein fragment composition is shown as SEQ ID No.1-3. The characteristic protein fragment composition or the mass spectrum model can be usedfor diagnosis and screening of the thalassemia and evaluation of the treatment method of the thalassemia and the therapeutic effect of the drug. The method is simple and easy to operate and is high in accuracy, and a new method and idea is provided for diagnosis and screening of the thalassemia, the treatment method and drug therapeutic effect evaluation.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a composition or a mass spectrometry model for detecting characteristic hemoglobin of thalassemia, and using the composition or the model, rapid identification of abnormal hemoglobin and detection of characteristic proteins, especially diseases related to hemoglobin fragments Efficacy evaluation of therapeutic methods or drugs. At the same time, the present invention also provides diagnostic products and applications related to the composition or model. Background technique [0002] Hemoglobin, also known as hemoglobin, is a heterotetrameric complex composed of two β gene globulins and two α gene globulins. It is the main component of red blood cells and can combine with oxygen to transport oxygen and carbon dioxide. The hemoglobin level can well reflect the degree of anemia. Hemoglobinopathies are the largest group of hereditary human single-gene diseases in the world. They are also...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/62G01N33/68G01N33/72A61K49/00
CPCG01N33/6848G01N33/721A61K49/0004G01N2333/805
Inventor 何昆吕倩安鹏牛燕燕马庆伟向华
Owner 长沙湘华质谱医学科技有限公司
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