Expert Marine Lipopolysaccharide (LPS) Analysis: Fueling Scientific Breakthroughs
At CD BioGlyco, our service system starts with the extensive Marine Carbohydrate Characterization Services that aim to fully analyze the structure and function of marine carbohydrates, providing a solid foundation for marine drug development, material science, and environmental research.
For further refinement, we specifically provide Marine Microbial Polysaccharide Characterization Services, focusing on the characterization of polysaccharides isolated from the unique resource of marine microorganisms. Among this series of services, the LPS characterization service is a highlight. Our LPS characterization service uses cutting-edge analytical technology to conduct comprehensive and in-depth characterization from chemical composition, molecular weight, and structural characteristics to biological activity.
Extraction and Purification
Before any characterization begins, one of the critical steps is the extraction and purification of LPS from marine bacterial sources.
- Hot phenol-water extraction: This method utilizes hot phenol to separate LPS from the bacterial cell wall, ensuring minimal contamination from proteins and nucleic acids.
- Ultra-centrifugation: Following extraction, ultra-centrifugation helps purify the LPS further by removing any remaining cellular debris.
Structural Analysis
- Mass spectrometry (MS): We use MALDI-TOF and ESI-MS to provide detailed information about the molecular weight of LPS components and their structural intricacies.
- Nuclear magnetic resonance (NMR) spectroscopy: This method helps us elucidate the detailed carbohydrate structure, including monosaccharide composition and linkages.
- Gas chromatography-mass spectrometry (GC-MS): For fatty acid profiling, GC-MS is employed after methanolysis to understand the lipid A structure of the LPS.
Chemical Composition Analysis
- Monosaccharide analysis: We conduct acid hydrolysis followed by high-performance liquid chromatography (HPLC) or capillary electrophoresis to identify and quantify monosaccharides.
- Fatty acid analysis: Fatty acids are derivatized to methyl esters and analyzed using GC-MS to determine their composition and distribution within the LPS.
- Phosphate content: We assess the phosphate groups in LPS using colorimetric assays or NMR spectroscopy.
Functional and Biological Characterization
- Cell activation assays: We study the immunogenicity of marine LPS by conducting human or animal cell line assays to see how LPS activates the immune response.
- Limulus amebocyte lysate (LAL) assay: This assay helps determine the endotoxin activity of LPS, which is an indicator of its potential to trigger inflammatory responses.
- Binding studies: Using techniques like surface plasmon resonance (SPR) and isothermal titration calorimetry (ITC), we investigate the binding affinity of LPS to various biomolecules, such as proteins and receptors.
Molecular Modeling
To predict and visualize the structure-function relationship of marine LPS, we employ computational methods as follows.
- Molecular dynamics (MD) simulations: These simulations help in understanding the conformational dynamics of LPS in different environments, providing insights into their stability and interactions.
- Docking studies: We use molecular docking to hypothesize how LPS might interact with receptor molecules at the atomic level, aiding in the identification of potential binding sites and interaction energies.
Comparative and Evolutionary Studies
- Phylogenetic analysis: By comparing the LPS structures from various marine and terrestrial bacteria, we deduce evolutionary trends using bioinformatics tools.
- Comparative omics: Integrating data from genomics, proteomics, and lipidomics enables us to draw comprehensive comparisons and understand the unique features of marine LPS.
Publication
Technology: Chemical structure characterization of LPS
Journal: Marine drugs
IF: 6.085
Published: 2015
Results: The authors examined the chemical structure of LPS from two bacterial species associated with the sponge Suberites domuncula, specifically a commensal Endozoicomonas sp. and an opportunistic Pseudoalteromonas sp. Their electrophoretic patterns demonstrated distinct LPS structures for these bacteria. The electrospray ionization ion-trap mass spectra identified monophosphorylated molecules indicative of tetra- and pentaacylated structures, exhibiting common features between the two strains. Despite having unique structural characteristics, neither of the LPS from these bacteria affected the expression of the macrophage-expressed gene in Suberites domuncula, contrasting with the LPS from Escherichia coli.
Fig.1 ESI-mass spectrum obtained in the negative ion mode of lipid A from Pseudoalteromonas sp and ESI-mass spectrum obtained in the negative ion mode of lipid A from Endozoicomonas sp. (Gardères, et al., 2015)
Applications of LPS
- LPS can be used to study the model components of biomembranes due to its unique amphiphilic structure of lipophilic lipid A and hydrophilic oligosaccharide chains.
- LPS can be used to develop new biomaterials such as drug carriers and tissue engineering scaffolds due to its good biocompatibility and degradability.
- As one of the main components of bacterial cell walls, LPS can be used to trigger the host's immune response, including activating Toll-like receptors and inducing the production of cytokines.
Advantages
- We have advanced analytical technology and a professional technical team to perform high-precision structural analysis of LPS.
- We establish a comprehensive LPS bioactivity evaluation system. Through in vitro and in vivo experiments, we systematically evaluate the activity of LPS in immunomodulation, cell adhesion, and signal transduction.
- We have a professional data analysis team and an efficient data processing system to quickly and accurately process and analyze the large amount of data generated during the LPS characterization process.
- In the field of LPS characterization, we continue to pay attention to the latest scientific research trends and technological advances and are committed to transforming the latest research results into practical characterization technologies.
Frequently Asked Questions
What are the characteristics of marine LPS?
Marine LPS has unique biochemical properties, mainly including its complex molecular structure. It is usually composed of hydrophilic oligosaccharide chains and hydrophobic lipid A parts. This amphiphilic structure gives it good interfacial activity and biocompatibility. In addition, LPS exhibits a variety of biological activities, such as immunomodulation, anti-oxidation, cell adhesion, etc. These characteristics give it a wide range of potential applications in biomedicine, materials science, and environmental science. Its complex structure also makes it an ideal model for studying biomacromolecule interactions and cell signaling.
How is the customized characterization service carried out?
The customized characterization service is based on the specific needs of clients. First, we communicate in-depth with clients to understand their research objectives, sample characteristics, and expected characterization content. Subsequently, our technical team design a personalized experimental plan based on this information and execute it after confirming with the client. During the experiment, we maintain close communication with clients and provide timely feedback on experimental progress and results. Finally, we provide a detailed characterization report, including experimental methods, data results, and analysis discussions to meet the scientific research or industrial needs of clients.
CD BioGlyco provides clients with high-quality LPS characterization services with scientific and rigorous methods, comprehensive characterization strategies, and professional data analysis capabilities, helping them to successfully develop scientific research and industrial applications. Please feel free to contact us for more details if you are interested in our LPS characterization service!
Reference
- Gardères, J.; et al. Lipopolysaccharides from commensal and opportunistic bacteria: characterization and response of the immune system of the host sponge Suberites domuncula. Marine drugs. 2015, 13(8): 4985-5006.
