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Development of a water-in-oil-in-water multiple emulsion system integrating biomimetic aqueous-core lipid nanodroplets for protein entity stabilization. Part II: process and product characterization

作者： Cássia A. Glasser¹, Marta M.D.C. Vila^1,2, Júlio C. Pereira³, Marco V. Chaud¹, José M. Oliveira Júnior¹

¹Labnus – Biomaterials and Nanotechnology Laboratory, i(bs)2

- intelligent biosensing and biomolecule stabilization

Research Group, University of Sorocaba, Sorocaba/SP, Brazil.

²Institute of Chemistry, University of Campinas, Campinas/SP, Brazil.

³Department of Environmental Sciences, Federal University of São Carlos, Sorocaba/SP, Brazil.

⁴CEB - Centre of Biological Engineering, University of Minho, Braga, Portugal.

摘要：The aqueous-core enclosed in lipid nanoballoons integrating multiple emulsions of the type water-in-oil-in-water mimic, at least in theory, the environment within viable cells, thus being suitable for housing hydrophilic protein entities such as bioactive proteins, peptides and bacteriophage particles. This study reports a complete physicochemical characterization of optimized biomimetic aqueous-core lipid nanoballoons housing hydrophilic (BSA) protein entities, evolved from a statistical 23×31 factorial design study (three variables at two levels and one variable at three levels) that was the subject of the first paper of a series of three, aiming at complete stabilization of the three-dimensional structure of protein entities attempted via housing the said molecular entities within biomimetic aqueous-core lipid nanoballoons integrating a multiple (W/O/W) emulsion. The statistical factorial design followed led to the production of an optimum W/O/W multiple emulsion possessing quite homogeneous particles with an average hydrodynamic size of (186.2 ± 2.6) nm and average Zeta potential of (−36.5 ± 0.9) mV, and exhibiting a polydispersity index of 0.206 ± 0.014. Additionally, the results obtained for the diffusion coefficient of the lipid nanoballoons integrating the optimized W/O/W multiple emulsion were comparable and of the same order of magnitude (10−12 m2 s−1) as those published by other authors since, typically, diffusion coefficients for molecules range from 10−10 to 10−7 m2 s−1, but diffusion coefficients for nanoparticles are typically of the order of magnitude of 10−12 m2 s−1.