Empowering Research through Pristine Lipopolysaccharide (LPS) Production
LPS is a component of the outer wall of bacterial cell walls and is a substance composed of lipids and polysaccharides. LPS is also an endotoxin, which exhibits a wide range of biological activities when it acts on the cells of other organisms such as humans or animals. As a company dedicated to research in glycobiology, CD BioGlyco has been drilling in the field of Marine Carbohydrate Production for many years and has mastered the Production of Marine Carbohydrates and Polysaccharide Purification Technology to provide high-quality marine polysaccharides production services to clients all over the world. Here, we provide LPS production services as shown below.
Extraction of LPS
LPS extracted by different methods may differ slightly in molecular mass and fine structure, CD BioGlyco provides suitable LPS extraction methods according to our clients' research needs. The extraction methods we provide include but are not limited to the following.
- Trichloroacetic acid (TCA) method
The TCA method is a common method to extract LPS from marine microorganisms at lower temperatures. LPS extracted using TCA has a high yield and good antigenicity.
- Phenol-water method
We utilize the phenol-water method to extract LPS from marine S-type bacteria and mutant R-bacteria. When LPS is extracted using phenol and water, the LPS dissolves in the aqueous phase, while most of the proteins are dissolved in phenol, and insoluble substances are present at the junction of the two phases. At the end of the extraction, the aqueous phase containing LPS is dialyzed and dried.
- Organic solvent extraction
LPS is often present in the outer membrane of the cell wall of microorganisms, so we use certain chemicals (e.g., phenol, chloroform, and petroleum ether) to release LPS by lysing the cell wall. In addition, since R-LPS has good solubility in organic solvents, this method can obtain high yields of R-LPS.
Purification of LPS
Extracted LPS can be contaminated by some small molecules such as polyamines, and cations (Ca2+, Mg2+, K+, etc.) in addition to proteins, nucleic acids, and polysaccharides. Therefore, further purification is required.
- Ultracentrifugation
Ultracentrifugation is a commonly used method for purifying LPS, which causes the LPS to precipitate, leaving the proteins, ribose, and other polysaccharides in the supernatant, which can be removed to obtain a more purified LPS.
- Electrodialysis (ECD)
ECD not only removes contaminants such as proteins, nucleic acids, and polysaccharides but also removes small molecules such as polyamines and divalent cations from LPS, resulting in high-purity LPS.
In addition to the above methods of LPS purification, we also offer chromatography, electrophoresis, and other methods to our clients.
Characterization of LPS
- Molecular mass determination
We offer high-performance liquid chromatography (HPLC) to analyze the molecular weight of LPS. In this method, we use glucose of different molecular weights as a standard and construct a linear equation based on the peaks in the spectra and use this to calculate the molecular weight of LPS.
- Structural characterization
We use infrared spectroscopy (IR), ultraviolet-visible spectroscopy (UV-Vis), and scanning electron microscopy (SEM) to characterize the structure and morphology of LPS.
- Monosaccharide composition analysis
The HPLC method is used to test the monosaccharide composition of LPS. LPS is hydrolyzed with acid then derivatized, and then analyzed using HPLC. Finally, the monosaccharide composition of LPS is calculated based on the area of the peaks in the spectra.
Publication Data
Technology: Limulus amebocyte lysate assay
Journal: Periodontology 2000
IF: 18.6
Published: 2022
Results: In this literature, the authors analyzed the structural relationships of LPS. The results of the research showed that LPS consists of three distinct structural domains, namely lipid A, core oligosaccharide, and O-specific sugar chain. Lipid A contains four acyl chains and secondary fatty acids directly attached to the glucosamine backbone, and structurally distinct lipid A can completely block almost all pro-inflammatory responses by competitively binding to the corresponding host receptor. In addition, the authors quantified and successfully detected LPS activity using the Limulus amebocyte lysate assay.
Fig.1 The structure of LPS. (Pussinen, et al., 2022)
Applications
- LPS is a bio-macromolecular reagent, and LPS production services can be used in the process of immunomodulation assays in drug discovery and development, and the testing of the anti-inflammatory activity of drugs.
- LPS can be used to stimulate immune cells such as monocytes and macrophages to trigger an inflammatory response, so LPS production services can be used to explore the mechanisms of immune cell activation and cytokine secretion.
- LPS is a major component of the cell membrane of most Gram-negative bacteria, so LPS production services help scientists investigate the structure and biological functions of bacteria.
Advantages
- Our service team has a deep background in marine biomolecular research and is skilled in the production of marine microbial polysaccharides, providing custom LPS production solutions to our clients.
- With first-class chemical and biological technologies, we provide one-stop solutions for the production, purification, characterization, and bioactivity analysis of LPS to our clients.
- We not only provide our clients with high-quality LPS production services but also offer LPS Products from different sources and through different purification methods.
Frequently Asked Questions
What are the biological functions of LPS?
LPS can induce host cell-intrinsic immune response and play a very important role in the recognition, adhesion, transfer, and pathogenesis of bacteria. Its structural composition is closely related to the serotype and pathogenicity of bacteria. When released in high doses, LPS can cause fever, microcirculatory disorders, endotoxic shock, and disseminated intravascular coagulation. Low doses of LPS can lead to the production of cytokines such as tumor necrosis factor TNF-α and interleukin IL-6, stimulate Toll-like receptor TLR4, help the host to resist infection, and can be used as an active ingredient in vaccines and adjuvants.
Why LPS is negatively charged?
LPS has sugar chains bound by glycosidic bonds, and electrostatic repulsion occurs between groups with the same charge on the sugar chains, resulting in a negative charge on the LPS.
CD BioGlyco is a company dedicated to research in glycobiology, offering a wide range of LPS production and analysis to our clients. We have been drilling in the field of marine carbohydrate production for many years and have earned praise from all over the world. Please feel free to contact us if you would like to know more about LPS.
Reference
- Pussinen, P.J.; et al. Periodontitis and cardiometabolic disorders: The role of lipopolysaccharide and endotoxemia. Periodontology 2000. 2022, 89(1): 19-40.
