CD BioGlyco has provided reliable and cost-effective Lipidomics Analysis Services to clients all over the world. We will adhere to the principle of customer-centricity, keep full transparency in the experimental process and speed up the research of Sphingolipid Analysis for clients.
Gangliosides participate in cellular proliferation, differentiation, adhesion, signal transduction, cell-to-cell interactions, tumorigenesis, and metastasis. The accumulation of gangliosides has been linked to several diseases including Tay-Sachs and Sandhoff disease. An autoimmune response against gangliosides can lead to Guillain-Barre syndrome. GM stimulates neuronal sprouting and enhances the action of nerve growth factor (NGF) by directly and tightly associating with Trk, the high-affinity tyrosine kinase-type receptor for NGF. It is the specific cell surface receptor for cholera toxin.
Fig.1 The ganglioside biosynthetic pathway and related pathologies (Sipione, et al., 2020).
CD BioGlyco provides reversed-phase chromatography coupled with mass spectrometry for the structural study and determination of GM. Taking the sea urchin sample as an example, the analysis process is as follows.
The lyophilized powder of sea urchin ovary is extracted twice with chloroform-methanol-water. The mixture is then stirred at room temperature for 30 min, then centrifuge, filter, and pool the supernatant. Adding water to the supernatant to adjust the final ratio of chloroform-methanol-water. After vortexing and centrifugation, the resulting upper phase is passed through the column for cleaning and desalting. The recovered GM is redissolved in chloroform-methanol-water and eluted on the column. After washing the column with solvent A to remove neutral lipids, a linear gradient from solvent A to solvent B is performed.
MS experiments to study GM cleavage pathways are performed on a triple quadrupole instrument. ESI-MS employs both positive and negative modes. MS2 spectra are acquired from collision-induced dissociation (CID) at a pressure of 0.2 MPa using nitrogen as the collision gas.
The liquid chromatography system is equipped with a binary pump. Chromatographic separation is carried out at room temperature. The mobile phase prepared by mixing acetonitrile and ammonium acetate in a certain ratio is used to elute sea urchin GM containing different ceramides.
To analyze the ceramide structure and relative content of each molecule in sea urchin GM, MRM analysis is performed by monitoring sialic acid ions in negative ion mode and long-chain bases (LCB) ions in positive ion mode. During the MRM analysis, precursor ions of the sialic acid ion in negative ionization mode and precursor ions of the LCB ion in positive ionization mode are monitored.
CD BioGlyco provides high-quality GM analysis service, including GM1, GM2, GM3, and GM4 analysis services.
CD BioGlyco has developed highly specialized platforms and various systematic solutions for Ganglioside Analysis. We provide reversed-phase chromatography coupled with mass spectrometry technology for GM Analysis. If you are interested in our services, please contact us for more details without any hesitation.
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