Driving Innovation with Tailored Aplysianin Solutions for A Sustainable Future
Sea hares, opisthobranch mollusks that have piqued the curiosity of numerous researchers exploring biologically potent compounds, primarily harbor low molecular weight substances from their algal diets. At CD BioGlyco, we utilize diverse technologies to produce three kinds of aplysianins, comprising aplysianin A, aplysianin E, and aplysianin P, each of which plays a pivotal role in enhancing biotechnological advancements.
- Aplysianin A: We will produce aplysianin A by isolating it from the sea hare Aplysia kurodai's albumen gland. It displays antibacterial and antineoplastic properties.
- Aplysianin E: We isolate aplysianin E, an antitumor factor, inducing tumor lysis, from the supernatant of a homogenate of eggs of the sea hare A. kurodai.
- We perform isolation and characterization of a novel cytolytic factor, aplysianin P, from the purple fluid of A. kurodai.
Our aplysianin production encompasses a series of specific steps:
Collection of A. kurodai
Our process begins with the acquiring of living A. kurodai, and eggs of A. kurodai, that are high-quality, laying the foundation for the entire production journey.
Pretreatment
- We will extract the anterior genital mass, which consists of the globular albumen gland and surrounding mucous gland, from freshly dissected animals for the isolation of aplysianin A.
- Egg masses of A. kurodai will be homogenized with saline, and the homogenate will be centrifuged. The resulting supernatant is then recentrifuged to obtain a clear supernatant, which serves as our starting material for the purification of aplysianin E.
- By irritating the animals, we obtain a purple fluid, which we freeze at -20°C for storage, for producing aplysianin P.
Extraction and purification
During this production, we mainly use ion exchange chromatography and gel filtration techniques for purification.
- Firstly, the raw materials after pretreatment undergo dialysis against a phosphate buffer before being applied to columns.
- The columns will be washed with the same buffer and then eluted using a sodium chloride gradient.
- Fractions exhibiting cytolytic activity will be combined, concentrated, and passed through a membrane before being further separated on a column.
- Active fractions will be concentrated, purified through a column, and dialyzed against a phosphate buffer before being processed through a final column with a sodium chloride gradient for elution.
To ensure that the product meets quality standards and verify the effectiveness of the production process, we will conduct assays to accurately assess and confirm the performance and purity of aplysianins. Use various technologies to determine the content, including:
- High-performance liquid chromatography (HPLC)
- Sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE)
- Capillary electrophoresis (CE)
- Liquid chromatography with tandem mass spectrometry (LC-MS-MS)
Finally, the assay phase is conducted to evaluate the toxicity and bioactivity of the produced aplysianins.
Publication Data
Technology: Histological analysis, Cell culture, Western blot analysis, RNA isolation and quantitative real-time PCR, Cell proliferation assay, Annexin-FITC apoptosis assay, Statistical analysis
Journal: Oncotarget
Published: 2017
IF: 5.168
Results: In this study, the authors elucidated the contemporary landscape of breast cancer therapeutics and emphasized the pressing requirement for novel drugs with robust anti-tumor efficacy and minimal ancillary effects. As per ancient Chinese medicine records, aplysia species are noted for their efficacy in alleviating various forms of inflammation and exhibiting anti-tumor properties. The authors highlighted the significant anti-tumor activities of three compounds, namely aplysianin-A, aplysianin-E, and aplysianin-P, extracted from Aplysia kurodai. They conducted a series of experiments on the other Aplysia kurodai extract, aplysin. It is a bromosesquiterpene with a molecular weight of 295, isolated by Japanese researchers in 1963. The results showed the efficacy of aplysin in halting the proliferation of breast tumors through robust suppression of the PI3K/AKT pathway, particularly targeting the downstream FOXO3a cascade. These investigations lay a solid groundwork for further exploration into the therapeutic potential of aplysin, especially in conditions where PI3K/AKT serves as a key oncogenic driver, such as breast, colon, ovarian, prostate, and pancreatic cancers.
Applications
- Aplysianin is extensively utilized for its inhibitory effects on various bacteria.
- Within the domain of drug development, there exists the potential for aplysianin to be applied, with a focus on investigating prospective treatments for cancer.
- For biomedical research, aplysianin is employed as an experimental instrument to delve into the repercussions of protease inhibitors on cells and organisms.
Advantages
- Each step in our process is meticulous, from the acquisition of premium-quality A. kurodai raw material to the final characterization and assay.
- By employing a comprehensive array of sophisticated techniques such as capillary electrophoresis, SDS-PAGE, LC-MS-MS, and HPLC for characterization, we uphold stringent quality standards of our aplysianin products.
- We not only guarantee the safety and efficacy of our aplysianins but also validate their diverse applications in biomedical research and drug development.
Frequently Asked Questions
Apart from those mentioned above, what other technologies do we have for characterization?
- Nuclear magnetic resonance (NMR)
- Atomic force microscopy
- Fourier transform Raman (FT-Raman)
- Thermogravimetric analyzer (TGA)
- Energy dispersive X-ray spectroscopy (EDS)
- Gas chromatography (GC)
- X-ray diffraction (XRD)
What other Marine Biomolecules can we produce?
CD BioGlyco offers a comprehensive Glycoprotein Production Service, including precise steps such as collection, pretreatment, extraction, purification, characterization, and activity testing. With advanced and meticulous techniques, we ensure the production of high-quality, potent aplysianins for various biomedical applications. Please feel free to contact us to collaborate on driving innovation towards the future!
References
- From Wikipedia: https://en.wikipedia.org/wiki/Anaspidea#/media/File:Aplysia_californica.jpg.
- Zhang, X., et al. Breast cancer suppression by aplysin is associated with the inhibition of the PI3K/AKT/FOXO3a pathway. Oncotarget. 2017, 8(38): 63923.
