Unleashing The Power of Hemocyanin for A Brighter Tomorrow!
CD BioGlyco’s research team offers production services for various Marine Biomolecules, including Marine Carbohydrates, Marine Glycoproteins, Marine Lectins, Marine Glycosides, and Marine Glycolipids. Through our various advanced technologies, we provide professional production services for hemocyanin, a crucial protein found in molluscs and arthropods that functions as the primary oxygen carrier in their hemolymph. Within creatures such as arthropods, molluscs, and specific insect larvae, hemocyanins exist as substantial extracellular proteins. These entities possess an electrostatic surface predominantly characterized by negative charges, a feature that enhances the solubility and hydration of hemocyanin in the hemolymph while preventing its interaction with negatively charged cellular surfaces like the glycocalyx. Hemocyanin derived from these organisms exhibits confirmed antiviral and antimicrobial properties, demonstrating utility in a wide array of scientific contexts.
These different types of hemocyanin have unique structures as they are produced by different organisms, and we provide tailored production services for each type to meet the specific requirements of our clients. We will produce various types of hemocyanin, including nautilus-type hemocyanin, squid-type hemocyanin, keyhole limpet-type hemocyanin, and mega-type hemocyanin. The main production process at CD BioGlyco includes the following steps:
Selection of hemocyanin type and raw material
Depending on the client’s specific requirements, we will provide appropriate advice to select a suited hemocyanin type. Based on the chosen type, we will acquire the relevant marine biological materials to serve as the source for hemocyanin production.
Sample pretreatment
We will filter and then treat the raw material with phenylmethanesulphonyl fluoride (PMSF) to prevent proteolysis. After removing cells by centrifugation, hemocyanin will be concentrated using a tangential ultrafiltration system. Sucrose is added as a cryoprotectant before storing the hemolymph at -20°C.
Extraction
Our research team employs the preparative ultracentrifugation method and salting-out procedure to isolate rude hemocyanin.
- The hemocyanin will be isolated from the concentrated sample by precipitation at a specific saturation level with crystalline ammonium sulfate, followed by centrifugation.
- Besides, the hemocyanin pellet will be suspended in a buffer and dialyzed to remove the ammonium sulfate.
- Subsequently, the material will be dialyzed against a stabilizing buffer and the hemocyanin solution will be clarified by centrifugation.
Purification
The extracted hemocyanin is purified through gel filtration chromatography to remove impurities and isolate the protein.
Analysis
The purified hemocyanin is subjected to rigorous quality control tests and analysis to ensure its high quality.
- We utilize the absorption spectra for measurement of the protein concentration based on an absorption coefficient.
- In transmission electron microscopy, the sample will be adsorbed to a glow-discharged support film, stained with ammonium molybdate, and viewed under specified conditions.
- We will perform sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) in polyacrylamide gel using Laemmli's method, and utilize protein standards for determining molecular masses.
By following these steps, we ensure the efficient and high-quality production of various types of hemocyanin for diverse applications.
Publication Data
Technology: Transmission electron microscopy, Gel filtration chromatography, Tumor model establishment, Flow cytometry analysis, enzyme-linked immunosorbent assay (ELISA)
Journal: BMC immunology
Published: 2014
IF: 3.0
Results: In the study, authors isolated and purified hemocyanins from the marine snail Rapana thomasiana (RtH) and terrestrial snail Helix pomatia (HpH). Transmission electron microscopy was used to conduct structural studies, and the electron micrograph displayed didecamers typical of gastropodan hemocyanins. Then they tested the ability of HpH and RtH to induce apoptosis in C-26 cells through apoptosis assays, including studying phosphatidylserine surface expression and analyzing levels of induced apoptosis, necrosis, and Annexin-V/Propidium iodide staining at different time points. It was shown that no dose-dependent quantitative correlation was observed between the amount of hemocyanin added to the cells and the induced apoptosis across all time points tested. Additionally, the authors conducted phenotyping, cytokine detection, anti-C-26 antibody formation, survival analysis, and tumor model development. The results showed that hemocyanin-treated animals exhibited prolonged survival, inhibited tumor growth, splenomegaly, and reduced lung metastasis, indicating that hemocyanin can induce an anti-tumor immune response and moderate C-26 carcinoma symptoms.
Applications
- Hemocyanin serves as an effective carrier and is widely used in vaccine development and production to enhance vaccine immunogenicity and efficacy.
- Hemocyanin is being investigated as a candidate for antiviral and antibacterial drugs, showing potential for a wide range of biomedical applications.
- Hemocyanin may play a role in anti-tumor therapy, warranting further research in this area.
Advantages
- We offer tailored production services for different types of hemocyanin to meet the specific requirements of clients.
- Our research team provides appropriate advice on selecting the most suitable hemocyanin type to ensure optimal choices for production.
- Our researchers utilize various methods to perform rigorous quality control tests and analyses to ensure the high quality of the produced hemocyanin.
Frequently Asked Questions
What is the structure of hemocyanin?
Molluscan hemocyanins represent a category of substantial protein entities, showcasing molecular weights that spread over a vast range, approximately from 3.3 to 13.5 million daltons. These entities congregate in assemblies of ten units or larger, which are comprised of individual protein segments weighing between 330 to 450 thousand daltons each. Within the core of each subunit lies a complex arrangement of more than seven analogous functional components. The structural architecture of molluscan hemocyanins commonly manifests as hollow cylindrical constructs characterized by an exterior cylindrical casing and multiple internal collar regions situated along the inner circumference of the cylinder. The specific configuration through which the constituent units merge together varies contingent upon the particular functional makeup of the corresponding polypeptide chain's component units.
What are the effects of hemocyanin?
- Antibacterial properties
- Antifungal properties
- Anti-parasitic properties
- Antiviral properties
- Agglutinative properties
- Antitumor properties
The production process for hemocyanin involves the selection of hemocyanin type and raw material, sample pretreatment, extraction, purification, and analysis. CD BioGlyco is dedicated to innovation and research in glycoprotein production. Through continuous research and development efforts, we persistently enhance efficiency, quality, and effectiveness in producing carbohydrate products. If you would like to contribute to scientific advancements and innovation in various fields, please feel free to contact us! CD BioGlyco is a reliable partner for seeking customized solutions and high-quality hemocyanin products.
Reference
- Gesheva, V.; et al. Anti-cancer properties of gastropodan hemocyanins in murine model of colon carcinoma. BMC immunology. 2014, 15: 1-11.
