Method for analyzing glucoprotein

A technology for glycoproteins and binding proteins, which is applied in the field of glycoprotein analysis, can solve the problems of long cycle time, cumbersome detection steps, and low detection throughput, and achieve the effects of small sample volume, wide application range, and good repeatability

Active Publication Date: 2008-11-19
SHANXI LIFEGEN
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Problems solved by technology

[0018] The purpose of the present invention is to provide a method for analyzing glycoproteins, which solves the te

Method used

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Embodiment Construction

[0045] The lectin chip is a high-throughput technology for detecting glycoproteins. The present invention uses a lectin chip combined with surface-enhanced laser desorption ionization time-of-flight mass spectrometry technology to detect, analyze, and identify glycoproteins, and compare the glycoproteins in different biological samples. Content and structural differences, research and identification of biomarkers. The specific implementation steps are as follows:

[0046] (1) Preparation of lectin chips:

[0047] (1.1) Spotting the chip: Dissolve various lectins or sugar-binding proteins in the spotting buffer to make a spotting buffer of 1-5 mg / mL; spot the spotting buffer on a chip spotting instrument On the derivatized solid phase carrier; incubate at 25-60°C for 12 hours to obtain a lectin chip. The spotting buffer can be phosphate buffer, carbonate buffer, Tris-HCl buffer, Bis-tris buffer or Hepes buffer, etc. The derivatized solid phase support can use oxirane groups,...

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Abstract

Disclosed is a method for analyzing glycoprotein, which comprises steps of firstly preparing a spotting buffer solution, spotting a chip, closing, cleaning and then drying, finally obtaining an agglutinin chip. A biological specimen is directly added on the agglutinin chip for undergoing steps of incubation, cleaning for removing hybridprotein, forming a crystal and dissociating the crystal, a time-of-flight mass spectrometer is used for detecting the flying time of charged ions in a vacuum electric field so as to obtain the glycoprotein graphical spectrum information, visually display the glycoprotein information such as molecular weight and abundance in the detected biological specimen, and identify the glycoprotein differential expression. Comparing the glycoprotein graphical spectrum information of the detected biological specimen with the sample graphical spectrum information can obtain the glycoprotein information of the differential expression. The method solves the technical problems of low detection flux, complicated detection steps and long cycle in the background technique. The method has advantages of convenient operation, high sensitivity and good repeatability, and is applied to the fast analysis and application in clinic medicine.

Description

technical field [0001] The invention relates to a method for analyzing glycoproteins, in particular to a method for detecting and identifying glycoproteins by using a lectin chip combined with a surface-enhanced laser analysis ionization time-of-flight mass spectrometry technique. Background technique [0002] Carbohydrates are the main source of energy required by organisms to maintain life activities, the basic raw materials for organisms to synthesize other compounds, and the structural raw materials for organisms. They are one of the biological macromolecules involved in the essence of life activities. Sugars have multiple hydroxyl groups, and glycosidic bonds are divided into α and β configurations. The connection of monosaccharides may produce a large number of isomers. Therefore, the sugar chain structure contains very rich biological language information and is a high-density structure. information carrier. Cells are the basic unit of living organisms, and a wide va...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N27/64
Inventor 李铮陈超简强王婷孙士生王芬
Owner SHANXI LIFEGEN
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