Gangliosides Analysis

What are Gangliosides?

Gangliosides are a type of glycosphingolipid, which are complex molecules composed of a lipid (fat) and a carbohydrate (sugar) component. They are found predominantly in the nervous system, particularly in cell membranes, where they play important roles in cell-cell communication and signaling.

Structurally, gangliosides consist of a ceramide lipid tail linked to a complex oligosaccharide chain. The carbohydrate portion of gangliosides contains sialic acid residues, which are negatively charged and contribute to the overall negative charge of the cell membrane. This negative charge influences cell-cell interactions and membrane stability.

Gangliosides are particularly abundant in neuronal membranes, where they are involved in numerous cellular functions, including cell adhesion, signal transduction, and modulation of ion channels. They are also implicated in processes such as neuronal development, synaptic transmission, and neuronal plasticity.

In addition to their roles in the nervous system, gangliosides are also found in other tissues and organs, albeit in lower concentrations. They have been implicated in various physiological and pathological processes, including immunity, inflammation, cancer metastasis, and certain neurodegenerative diseases.

At Creative Proteomics, we offer a range of tailored ganglioside analysis services to meet different research needs for accurate characterization and quantification of gangliosides.

Gangliosides Analysis Services by Creative Proteomics

Gangliosides Qualitative and Quantitative Analysis: Utilizing high-resolution mass spectrometry, we offer precise identification and quantification of various gangliosides species in complex biological systems.

Gangliosides Structural Elucidation: Through advanced MS/MS techniques, our services provide in-depth structural characterization of gangliosides, enabling insights into their functional roles.

Gangliosides Profiling and Mapping: We deliver comprehensive gangliosides profiling to map their distribution across different tissues and sub-cellular locations.

Bioinformatics Integration: Our analysis includes advanced bioinformatics support to interpret the complex data associated with gangliosides research, facilitating a deeper understanding of their biological implications.

Analytical Techniques for Gangliosides Analysis

• High-Resolution Mass Spectrometry (HR-MS): Using instruments such as the Thermo Scientific™ Q Exactive™ HF-X Hybrid Quadrupole-Orbitrap™ Mass Spectrometer, we provide unparalleled precision in the detection and quantitation of gangliosides.
• Liquid Chromatography-Mass Spectrometry (LC-MS/MS): Our Agilent 6545XT AdvanceBio LC/Q-TOF system offers superior resolution and sensitivity for the detailed structural analysis of gangliosides.
• Shotgun Lipidomics: Incorporating this high-throughput technique enables a comprehensive and unbiased profiling of gangliosides directly from complex biological samples.

Sample Requirements for Gangliosides Analysis

Deliverables for Gangliosides Analysis

Comprehensive Analytical Reports: Detailed summaries of experimental procedures, methodologies, and key findings, providing a clear understanding of the results and their relevance to the research goals.

Ganglioside Profiling Data: Detailed profiles of detected ganglioside species, including identification and quantification results, facilitating comparative studies and biomarker discovery.

Structural Characterization Results: In-depth analysis of ganglioside structures, including glycan moiety composition, ceramide tail structures, and sialic acid linkage patterns, offering insights into their molecular complexity and functional implications.

Quantitative Analysis Data: Precise measurement of ganglioside levels in biological samples, enabling accurate assessment of their concentrations under various conditions, aiding in the identification of potential therapeutic targets.

Customized Consultation Services: Personalized support from experienced scientists throughout the analysis process, including tailored solutions and actionable insights specific to the research objectives.

Applications of Gangliosides Analysis

Neurodegenerative Disease Research: Facilitates understanding and identifying biomarkers for diseases like Alzheimer's and Parkinson's.

Rare Disease Studies: Assists in the study and development of treatments for disorders such as GM1 and GM2 gangliosidoses.

Cancer Research: Aids in understanding the role of gangliosides in tumor progression, metastasis, and angiogenesis.

Developmental Biology: Provides insights into the roles of gangliosides during brain development and neurogenesis.

Pharmacology and Toxicology: Evaluates the impact of drugs on ganglioside metabolism and elucidates potential toxicological effects.

Title: Characterization of Ganglioside Expression Profiles in Neuroblastoma Cell Lines Using LC-MS and HPTLC Analysis

Background

Gangliosides, consisting of sialic acid-containing oligosaccharide chains linked to ceramide anchors, play crucial roles in cellular processes and have been implicated in neuroblastoma (NB) pathogenesis. Understanding ganglioside expression profiles in NB cell lines could provide insights into tumor behavior and potential therapeutic targets.

Sample

Eleven NB cell lines were used in the study, including those with and without MYCN gene amplification. Cell lines were cultured under standard conditions in RPMI-1640 supplemented with 10% FBS.

Technical Methods

Cell Culture and Antibodies: NB cell lines were obtained from cell banks or as a gift and cultured in RPMI-1640 with 10% FBS.

Lipid Extraction: Lipids were extracted from cell pellets using chloroform/methanol and chloroform/isopropanol/water, desalted, and analyzed by HPTLC and LC-MS/MS.

HPTLC Analysis: Lipids were separated on Silica gel plates, stained, and immunostained for ganglioside detection. Acetylated GD2 was detected after chromatography and deacetylation.

LC/ESI-MS Analysis: Lipids were separated by high-performance LC, and MS/MS analysis was performed to identify ganglioside molecular species.

Reverse Transcription-PCR (RT-PCR) Analysis: Total RNA was extracted, cDNA synthesized, and RT-PCR performed to analyze gene expression related to ganglioside biosynthesis pathways.

Clustering Analysis: Ganglioside expression profiles were used for hierarchical clustering analysis using R software.

Results

LC-MS Analysis: Ganglioside expression profiles varied among NB cell lines, with differential expression of pathway a (e.g., GD1a) and pathway b (e.g., GD2) gangliosides. O-acetylated gangliosides were detected in all cell lines.

HPTLC Analysis: Ganglioside expression profiles detected by HPTLC were consistent with LC-MS findings for major gangliosides.

TLC Immunostaining: Differences in GD2 expression among NB cell lines were confirmed by TLC immunostaining.

Clustering Analysis: NB cell lines were classified into three groups based on their ganglioside expression profiles.

Correlation with Gene Expression: Ganglioside expression profiles were correlated with the expression of ganglioside synthase mRNA and neural differentiation markers, suggesting a biological significance of ganglioside expression in NB pathogenesis.

The LC-MS analysis of gangliosides extracted from the CHP134 cells.

Reference

1. Kaneko, Tomonori, et al. "Neuroblastoma cells can be classified according to glycosphingolipid expression profiles identified by liquid chromatography-tandem mass spectrometry." International journal of oncology 37.5 (2010): 1279-1288.