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Glycan analysis method

An analysis method and technology of sugar chains, applied in the field of analysis of sugar chain structures, can solve the problems of pre-processing cost, expensive, and complicated costs, and achieve low-cost effects

Active Publication Date: 2019-06-21
SHIMADZU SEISAKUSHO CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0021] As described above, in the existing sugar chain analysis methods, there is a problem that in order to reliably negatively ionize the sugar chains to be analyzed and obtain MS / MS spectra reflecting information on their branched structures, expensive special substrates are used instead of Common matrices that are easily purchased, etc., and pretreatment for sample preparation is costly and complicated

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  • Glycan analysis method
  • Glycan analysis method
  • Glycan analysis method

Examples

Experimental program
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experiment example 1

[0069] As a sample, an unlabeled NA2-type standard sugar chain manufactured by SIGMA-ARDRICH, USA was used. The chemical formula of this sugar chain is the same as that shown in Comparative Example 1. This standard sugar chain is dissolved in a liquid matrix containing 2AA, which is also a labeling reagent, as an acidic group-containing organic substance, and the reaction between the sugar chain and 2AA is promoted by heating to form a sugar chain labeled body. More specifically, add 5 μL of tetramethylguanidine (TMG=1,1,3,3,-Tetramethylguanidine) stock solution to 45 μL of 1M 2AA solution and dissolve it, collect 5 μL and dilute with 50% ACN 45 μL , The resulting substance is used as a matrix solution (liquid matrix). 0.5 uL of the matrix solution and 1 uL of the sample solution were mixed on a MALDI sample plate (the above-mentioned μFocus MALDI plate manufactured by Hudson Surface Technology Co., Ltd.), and directly heated together with the plate on a heating block at a tem...

experiment example 2

[0077] In order to label sugar chains, 5-aminosalicylic acid (5ASA) was used instead of 2AA, except that the measurement samples were prepared in exactly the same procedure as in Experimental Example 1 above, and MS / MS analysis was performed in negative ion mode. The MS / MS spectrum thus obtained is shown in Figure 5 . It is the deprotonation detected at m / z1774.7 ([M-H] - ) The result of MS / MS analysis performed on the ion as the precursor ion.

[0078] Peaks of approximately the same pattern as in Experimental Example 1 are observed, that is, at the same mass-to-charge ratio as that of the MS / MS spectrum obtained in Experimental Example 1, it is observed that they originate from ring breaks caused by D ions, E ions, and GlcNAc residues. The peak of the A ion. From this, it can be seen that in the sugar chain analysis method of the present invention, as long as the sugar chains are the same, the MS / MS spectrum reflecting the same branch structure information can be obtained re...

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Abstract

A glycan having a branched structure is labeled using a labeling agent, such as 2-aminobenzoic acid, that has one site that is easily negatively charged. At this time, the reduction that is normally carried out in labeling through reductive amination is not carried out. A sample for mass spectrometry including the labeled glycan obtained in this way is prepared, and tandem mass spectrometry is carried out on the sample in a negative ion mode. The peaks of, for example, E ions and D ions reflecting the branch structure appear clearly in the tandem mass spectrum obtained in this way. As a result, structural analysis of the entirety of a glycan including a branched structure can be carried out easily.

Description

Technical field [0001] The present invention relates to a method for analyzing the structure of a sugar chain by mass spectrometry, and particularly to a method useful for analyzing the structure of a sugar chain having a branched structure (also called a tree structure). Background technique [0002] It is thought that more than half of the proteins constituting the organism are modified by sugar chains, and sugar chain modifications play an important role in the regulation of the structure and function of the protein. In addition, through recent studies, it is known that sugar chains expressed on cell surfaces are involved in various in vivo phenomena such as cell-cell interaction, signal transmission, development / differentiation, fertilization, and cancer metastasis. [0003] The sugar chain of a modified protein rarely has a structure in which monosaccharides are connected in a straight chain, and in most cases has a branched structure. The structure of this branched sugar cha...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/62
CPCG01N33/6848G01N33/58G01N2400/10H01J49/0027H01J49/004
Inventor 西风隆司
Owner SHIMADZU SEISAKUSHO CO LTD