Method and device for identifying glycopeptide segment

A glycosylation and peptide technology, which is applied in measuring devices, material analysis through electromagnetic means, instruments, etc., can solve the problems of many candidate results and low accuracy

Inactive Publication Date: 2011-05-25
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is to overcome the defects of too many candidate results obtained by the existing glyco

Method used

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  • Method and device for identifying glycopeptide segment
  • Method and device for identifying glycopeptide segment
  • Method and device for identifying glycopeptide segment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1 Device for identifying glycosylated peptides

[0058] combined with image 3 , the device consists of:

[0059] (1) a mass spectrum filtering unit, which selects a mass spectrum peak with a sufficiently high signal-to-noise ratio in the tandem mass spectrum mass spectrum of the glycosylated peptide sample;

[0060] (2) Obtain the single charge mass number unit: call the mass spectrum peak selected by the mass spectrum filter unit, if the source of the mass spectrum peak is a MALDI mass spectrum, then take the mass number of the mass spectrum peak as the single charge mass number, if the mass spectrum used for identification is an ESI mass spectrum, The mass number of the mass spectrum peak is deconvoluted to obtain its single charge mass number;

[0061] (3) Construct the mass number network unit: call the single-charge mass number obtained from the single-charge mass number unit, perform pairwise subtraction to obtain the mass difference number, and then us...

Embodiment 2

[0064] Example 2 Identification of glycosylated peptides of asialofetuin (Asialofetuin)

[0065] The glycosylated peptide of the standard glycoprotein asialofetuin (Asialofetuin) is used as an unknown sample for identification to test the feasibility and reliability of the method of the present invention.

[0066] experimental method:

[0067] 1. Sample preparation: dissolve the unknown glycoprotein sample in 25mM NH 4 HCO 3 Medium, heat denatured. After the solution was cooled, trypsin was added (according to the ratio of trypsin: glycoprotein sample mass ratio of 1:50), and the enzymolysis was carried out overnight at 37°C. The next day, the enzymatic hydrolysis solution was vacuum-centrifuged and dried for use in subsequent experiments.

[0068] 2. Mass spectrometry identification: AXIMA-QITTM MALDI TOF MS was used for tandem mass spectrometry identification. 2,5-dihydroxybenzoic acid (DHB) was dissolved in 30 vt% acetonitrile aqueous solution containing 0.1 vt% triflu...

Embodiment 3

[0094] Example 3 Identification of glycosylated peptides of horseradish peroxidase (HRP)

[0095] The glycosylated peptide of the standard glycoprotein horseradish peroxidase (Horseradish Peroxidase, HRP) is used as an unknown sample for identification to test the feasibility and reliability of the method of the present invention.

[0096] Mass spectrometer: LTQ-Orbitrap mass spectrometer

[0097] experimental method:

[0098] 1. Sample preparation: dissolve the unknown glycoprotein sample in 25mM NH 4 HCO 3 Medium, heat denatured. After the solution was cooled, trypsin was added (according to the ratio of trypsin: glycoprotein sample mass ratio of 1:50), and the enzymolysis was carried out overnight at 37°C. The next day, the enzymatic hydrolysis solution was vacuum-centrifuged and dried for use in subsequent experiments.

[0099] 2. Mass spectrometry identification: ESI-LTQ-Orbitrap XL Systems was used for tandem mass spectrometry identification, and the proteolytic pep...

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Abstract

The invention discloses a method for identifying a glycopeptide segment, which comprises the following steps: filtering a mass spectrogram; acquiring a single-charge mass number; constructing a mass number network; and retrieving a theory library. The invention also discloses a device for identifying the glycopeptide segment, which comprises a mass spectrogram filtering unit, a single-charge massnumber acquisition unit, a mass number network construction unit and a theory library retrieving unit. Compared with the conventional method for identifying the glycopeptide segment, the identification method and the identification device can efficiently acquire more accurate identification results in which false positive results are obviously reduced.

Description

technical field [0001] The invention relates to a method and device for identifying glycosylated peptides. Background technique [0002] With the development of high-throughput and high-resolution biological mass spectrometry (including matrix-assisted laser desorption ionization mass spectrometry (MAIDI-MS) and electrospray mass spectrometry (ESI-MS)), proteomics research based on biological mass spectrometry is also Significant progress has been made. At present, the top priority of proteomics research focuses on three main subfields in proteomics identification: qualitative identification, quantitative identification and protein modification identification. Among them, the identification of protein modification is an important factor that distinguishes proteomics from transcriptomics, so it has become the most advanced part of proteomics. [0003] The identification of protein modifications has been developed to this day, and relatively complete identification technique...

Claims

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Application Information

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IPC IPC(8): G01N27/62
Inventor 贺福初杨芃原张扬沈诚频刘铭琪陈瑶函
Owner FUDAN UNIV
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