Post-Translational Modification Analysis | Sugar Chain Structural Analysis by LCMS-IT-TOF

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Post-Translational Modification Analysis

Sugar Chain Structural Analysis by LCMS-IT-TOF

■ Analysis of Commercial Mouse Monoclonal Antibodies

Sugar chains, also called the "third chains of life," are attracting attention as biological information molecules. Sugar chains play important roles in biological phenomena such as intercellular signal transmission and molecular recognition and are known to be involved in cancer; chronic disease; infectious disease; immunological, brain, and developmental abnormalities; and degeneration.
The improved performance of mass spectrometers permits high-throughput sugar chain structural analysis, which previously required a long time.

The analysis of commercial mouse monoclonal antibodies is introduced below.
Test and research flow
1) Use SDS-PAGE to excise bands corresponding to antibodies from the gel.
2) In-gel digestion by PNGaseF.
3) Purification and labeling of sugar chains using the BlotGlyco kit (Sumitomo Bakelite).
4) MSn analysis by Prominence nano and LCMS-IT-TOF.

Fig. 1 shows the mass chromatograms ofm/z696.7672,m/z769.2833,m/z850.3369, andm/z930.9525.

Fig. 1 Prominence nano / LCMS-IT-TOF Mass Chromatograms

Fig. 2 to Fig. 5 show the MS/MS spectra using the precursor ions from the mass chromatograms in Fig. 1 above. The sugar chain structures shown in the diagrams were estimated using sugar chain structure prediction software.

High-Performance Liquid Chromatograph Mass Spectrometer

  • The LCMS-IT-TOF High-Performance Liquid Chromatograph-Mass Spectrometer produces MSn information with high mass accuracy that is ideal for sugar chain analysis.
  • A combination of the Prominence nano Nanoflow High-Performance Liquid Chromatograph with the NES-100 NanoESI Interface offers highly sensitive sugar chain analysis.
  • The high mass accuracy of the LCMS-IT-TOF allows highly accurate sugar chain estimation.
  • The neutral loss survey automatically evaluates preset specific neutral losses in the MS2 spectrum and runs MS3 measurements.
  • Dedicated software provides powerful support for the structural analysis of sugar chains.