Anammox bacteria wield an energy-efficient nitrogen metabolism enveloped in anammoxosome organelle composed of unique ladderane lipids. Thus, waste anammox biomass seems to be an attractive target for the isolation of ladderanes and subsequent production of artificial vesicles for drug delivery. This study proposed a novel method to isolate ladderane-rich anammoxosomes from aggregate mixed culture of Ca. Brocadia sapporoensis. Compared to conventional isolation protocols, the protocol was simplified by omitting the prepurification of anammox cells, replacing Percoll® with a sucrose gradient and prolonging the application of EDTA. This enhanced and simplified procedure efficiently removed EPS and other debris, thus yielding the layer of anammoxosomes as confirmed by control experiments and TEM. For the first time, the resulting ladderane isolates were used for the preparation of liposomes, both with and without the addition of pure dipalmitoylphosphatidylcholine (DPPC). Vesicles were successfully created, characterised by TEM and DLS, and anammox-based ladderanes were incorporated into their walls. These liposomes had interesting functional properties such as increased colloid stability at elevated concentrations, meaning a reduced tendency to form aggregates compared to model liposomes made solely of DPPC. Overall, this study offers insights into converting waste anammox biomass into a valuable resource for drug delivery.

• Keywords
• Ca. Brocadia sapporoensis, anammox bacteria, anammoxosomes, artificial liposomes, ladderanes,
• MeSH
• Liposomes * chemistry   metabolism   isolation & purification   MeSH
• Planctomycetes * MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Liposomes * MeSH

This study explores the potential of using liposomal electrokinetic chromatography as a ranking method for the rapid and simultaneous evaluation of drug-membrane interactions of a larger group of substances and assessing their sensitivity to tissue-specific parameters, namely pH, temperature, and lipid composition. We used a group of nine model drug substances to manifest how molecules could be classified for the relative sensitivity of drug-membrane interactions to pH and temperature. We observed that increasing the amount of liposomes in the background electrolyte significantly affected the separation kinetics of various active pharmaceutical ingredients, altering their mobility and/or peak shapes. Experiments with liposomes from bovine liver and heart tissue extracts revealed different interactions based on the lipid composition. Canagliflozin, which initially showed no electrophoretic mobility, migrated toward the anode in the presence of negatively charged liposomes. Mobility of positively charged substances, ambroxol and maraviroc, was suppressed by the interactions with liposomes. Their peaks also exhibited significant tailing. The effect on the separation of negatively charged compounds was significantly weaker. A small change in mobility was observed only in the case of deferasirox. We also examined the effect of temperature during separation, and we observed that increased temperature generally enhanced effective mobility due to lower electrolyte viscosity and increased lipid bilayer fluidity. Lastly, we tested the effect of sodium phosphate buffer pH (ranging from 6.0 to 8.0) with 4% liposomes on drug-liposome interactions. However, the effects were complex due to changes in API ionization and liposome surface charge, complicating the distinction between pH effects and liposome presence on API behavior. Our findings emphasize the significance of liposome composition, temperature, and pH in studying the interactions of liposomes with drugs, which is crucial for optimizing liposome-based drug delivery systems.

• Keywords
• Active pharmaceutical ingredients, Capillary electrophoresis, Interactions, Liposomal electrokinetic chromatography, Liposomes, Pseudostationary phase,
• MeSH
• Chromatography, Micellar Electrokinetic Capillary * methods   MeSH
• Hydrogen-Ion Concentration MeSH
• Pharmaceutical Preparations chemistry   MeSH
• Liposomes * chemistry   MeSH
• Cattle MeSH
• Temperature MeSH
• Animals MeSH
• Check Tag
• Cattle MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Pharmaceutical Preparations MeSH
• Liposomes * MeSH