Disialoganglioside 2 (GD2) Analysis Service
CD BioGlyco has been focusing on Lipidomics Analysis Services. Our Sphingolipid Analysis Services have been widely recognized by researchers, and we will continue to explore the needs of different clients.
Structure and Function of Disialoganglioside 2 (GD2)
Gangliosides are carbohydrate-containing sphingolipids that are ubiquitously expressed in normal tissues. However, GD2 isoforms have limited expression in normal tissues but are overexpressed in a wide range of tumors. GD2 is involved in tumor development and malignant phenotype by enhancing cell proliferation, motility, migration, adhesion, and invasion. For example, GD2, the major ganglioside present in human neuroblastoma cell lines, is abundantly synthesized by the vast majority of primary untreated neuroblastoma, and can be expressed in neuroblastoma regardless of disease stage. detected in patient plasma and tumor tissue. Faster progression and lower patient survival in neuroblastoma appear to be associated with higher circulating tumor-derived GD2 levels at diagnosis. Therefore, the analysis of GD2 is of great significance for tumor-related research.
Fig.1 Structure of GD2. (Wikipedia)
GD2 Analysis Service at CD BioGlyco
CD BioGlyco provides sensitive and reliable qualitative and quantitative Disialoganglioside (GD) Analysis Services for clients' samples. We offer an ultra-performance liquid chromatography (UPLC)/MS/MS method for the measurement of GD2. The method employs a simple protein precipitation strategy for sample extraction, chromatographic separation using UPLC, and detection in tandem Multiple Reaction Monitoring (MRM) modes to achieve high sensitivity and specificity for rapid analysis. Taking human plasma samples as an example, our analysis process is as follows.
- Sample extraction
Working stock solutions and plasma samples are first prepared. Methanol is added to each sample and vortex vigorously. Then, the spiked sample is centrifuged to pellet the protein. Transfer the supernatant to an autosampler vial with a micro insert for LC/MS analysis.
- LC/MS/MS procedure
After ganglioside extraction, each sample is injected into a C18 UPLC column and eluted through an optimal gradient program. Introduce the LC eluate to waste during the first few minutes. Switch to the mass spectrometer after a certain period of elution, and at the same time, all monitored ganglioside components are gradually eluted. Afterward, the LC eluate is directed again to waste, during which time the column is cleaned and re-equilibrated for the next run. The column temperature is always controlled at 35°C during the analysis.
- Calibration curve
Quantification of GD2 in unanalyzed plasma and unknown plasma samples is implemented using standard addition calibration based on three levels, including one to-be-measured sample with nil addition and two standard addition calibrators. For each measured ganglioside specimen, the total peak area (TPA) is obtained by summing the individual peak area from every MRM transition channel in the corresponding sample from a single injection using the equation.
Advantages of Us
- The method employs a simple protein precipitation strategy for sample extraction.
- Detection in tandem MRM modes to achieve high sensitivity and specificity for rapid analysis.
- The UPLC/MS/MS method also be used to measure several common ganglioside species in human plasma, including GM2, GM3, and GD3.
- Comprehensive and reliable after-sales service.
CD BioGlyco has been working on Ganglioside Analysis Service for many years. We provide comprehensive GD2 analysis service through UPLC/MS/MS method. If you are interested in our services, please contact us for more details without any hesitation.
Reference
- Nazha, B.; et al. Disialoganglioside GD2 expression in solid tumors and role as a target for cancer therapy. Frontiers in Oncology. 2020, 10: 1000.
