Method for analyzing carbohydrate-binding protein by functional sugar chip and flight mass spectrometer

A technology for binding proteins and functional sugars, applied in the field of analyzing sugar-binding proteins, it can solve the problems of low detection throughput, long cycle, complicated detection steps, etc., and achieve the effects of good repeatability, small sample volume, and high-throughput analysis capability.

Active Publication Date: 2008-07-02
SHANXI LIFEGEN
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Problems solved by technology

[0011] The purpose of the present invention is to provide a method for analyzing sugar-binding proteins with a functional sugar chip and a flight mass

Method used

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Embodiment Construction

[0049] The realization steps of the present invention are as follows:

[0050] 1. Preparation of functional sugar chips:

[0051](1) Prepare the coupling agent solution and incubate the solid-phase carrier: dissolve the coupling agent in an organic solvent to prepare a 10-50 mol / L organic solution of the coupling agent; immerse the derivatized solid-phase carrier in it, and Incubate for 3 hours. The coupling agent of the present invention can be a coupling agent containing an amino group and a hydroxyl group, and the derivatized solid phase support can be a solid phase support derivatized with an oxirane group. Specifically, 4-hydroxybenzamide or 4-hydroxybutyric acid hydrazine can be used as the coupling agent. The coupling agent of the present invention may also be a coupling agent containing a carboxyl group and a hydroxyl group, and the derivatized solid phase support may also be a solid phase support derivatized with an amino group. According to the ratio of solute in ...

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Abstract

A method for analyzing carbohydrate-binding protein with a functional carbohydrate chip and a time-of-flight mass spectrometer comprises the following steps of: preparing a coupling agent solution, incubating solid carrier, cleaning, drying, dissolving reducing carbohydrate in water, adjusting to required concentration, spotting, and incubating to obtain a functional carbohydrate chip; directly adding a biological sample on the functional carbohydrate chip, incubating, removing impurities, forming crystal, dissociating crystal, detecting the flight time of charging ions in a vacuum electric field with a time-of-flight mass spectrometer, obtaining a protein map that directly shows molecular weight, abundance and other information of proteins in the detected biological sample, and identifying protein differential expression; and comparing the protein map information of the detected biological sample with a sample map information to obtain the information of carbohydrate-binding protein with differential expression. The invention solves the technical problems in the prior art, including low detection throughput, complex detection procedures, and long detection period. The invention has the advantages of simple operation, high sensitivity and good repeatability; and is suitable for rapid analysis in medical clinics.

Description

technical field [0001] The invention relates to a method for analyzing sugar-binding proteins, in particular to a method for detecting and identifying sugar-binding proteins by using a functional sugar chip combined with surface-enhanced laser analysis ionization time-of-flight mass spectrometry technology. Background technique [0002] At present, the techniques involved in the analysis and detection of proteomics mainly include: two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), electrospray mass spectrometry (ESI-MS), matrix-assisted laser desorption ion time-of-flight mass spectrometry (MALDI-TOF). -MS) and the recently emerging technology of protein chip coupled with time-of-flight mass spectrometry. Time-of-flight mass spectrometer is developed based on surface-enhanced laser desorption ionization spectrometry (surface-enhanced laser desorption ionization spectrometry, SELDI). In the same year, Bill Hutchens and Tai-Yung Yip of the United States proposed an...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N30/02
Inventor 李铮陈超南刚杨刚龙任挺立
Owner SHANXI LIFEGEN
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