Method for purifying interest protein in serum of patient with liver cancer through electrophoretic separation

Abstract

The invention discloses method for purifying interest protein in serum of a patient with a liver cancer through electrophoretic separation. The method comprises the steps of building a liver cancer and normal person diagnostic model and a liver cancer and chronic liver disease diagnostic model, and the protein in the serum of the patient with the liver cancer is purified through Tricine-SDS-PAGE electrophoretic separation; Blue 3GA and Protein A column materials are used for removing albumin and IgG in the serum of the patient with the liver cancer, then, a Tricine-SDS-PAGE electrophoretic gelresult shows that an ultralow molecular protein standard substance can extract six clear strips, the patient serum sample can have two strips through separation within the range of the molecular weight being 2,500-6,000 Da and the molecular weight being 8,000-11,000 Da, cutting gel enzymolysis is performed on target strips pointed by the red arrow in the picture, and the interest protein can be authenticated through the high performance liquid chromatography tandem mass spectrometry. On the basis of bioinformatics analysis, the effect of the key protein in a system signal network is analyzed;then, MRM absolute quantification verification is adopted for evaluating the liver cancer key protein polypeptide to diagnose the liver cancer value and analyze the clinic correlation, and the clinical application feasibility for liver cancer prevention, early diagnosis, prognosis and the like is put forward.

Description

technical field [0001] The invention relates to the fields of gene transcriptomics and proteomics, in particular to a method for separating and purifying the target protein in serum of patients with liver cancer by electrophoresis. Background technique [0002] In the 20th century, primary liver cancer has risen to become the second cancer killer in my country, second only to lung cancer in urban areas and gastric cancer in rural areas [1] . At present, about 35,000 people in Guangxi die of malignant tumors every year, among which, about 12,000 people die of liver cancer. It can be seen that liver cancer is the first cancer killer in Guangxi. In Guangxi, the hazards of liver cancer are very uneven. The counties and cities with high mortality rate of liver cancer are mainly concentrated in the southwestern part of Guangxi. The three counties of Fusui, Long'an and Wuming around Nanning have the highest incidence. The incidence density of liver cancer in this local area reach...

AI Technical Summary

Problems solved by technology

The high sensitivity of mass spectrometry can help people discover low-abundance proteins and peptides in biological samples in the stage of marker discovery. The dynamic range greatly affects the quantitative detection of low-abundance proteins and peptides in biological fluids by mass spectrometry

Although researchers have tried to use a variety of methods, including chromatographic separation as much as possible, removal of high-abundance proteins, etc., the precision and accuracy of the results are still far from meeting the requirements of analysis, until mass spectrometry multiple reaction monitoring (multiple reaction monitoring) The emergence of monitoring, MRM) technology

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