Purpose: To develop a general method for NP fabrication from various proteins with maintenance of biological activity. Methods: A novel general approach for producing protein nanoparticles (NP) by nanoprecipitation of the protein solutions in 1,1,1,3,3,3-hexafluoroisopropanol is described. Protein NP sizes and shapes were analyzed by dynamic light scattering, scanning electron and atomic force microscopy (SEM and AFM). Chemical composition of the NP was confirmed using ultraviolet (UV) spectroscopy, energy-dispersive X-ray spectroscopy (EDX) and circular dichroism (CD). Biological properties of the NP were analyzed in ELISA, immunofluorescent analysis and lysozyme activity assay. Results: Water-insoluble NP were constructed from globular (bovine serum albumin (BSA), lysozyme, immunoglobulins), fibrillar (fibrinogen) proteins and linear polylysines by means of nanoprecipitation of protein solutions in fluoroalcohols. AFM and SEM revealed NP sizes of 20–250 nm. The NP chemical structure was confirmed by UV spectroscopy, protease digestion and EDX spectroscopy. CD spectra revealed a stable secondary structure of proteins in NP. The UV spectra, microscopy and SDS-PAA gel electrophoresis (PAGE) proved the NP stability at +4°C for 7 months. Co-precipitation of proteins with fluorophores or nanoprecipitation of pre-labeled BSA resulted in fluorescent NP that retained antigenic structures as shown by their binding with specific antibodies. Moreover, NP from monoclonal antibodies could bind with the hepatitis B virus antigen S. Besides that, lysozyme NP could digest bacterial cellular walls. Conclusion: Thus, the water-insoluble, stable protein NP were produced by nanoprecipitation without cross-linking and retained ligand-binding and enzymatic activities.
Background: Synthesis of metal oxides nanoparticles with specific morphology and size has become the subject of many experimental protocols. Biosynthesis of the nanoparticles using plants is more preferable than physical and chemical methods because of its environmental friendliness.Objective: The purpose of this study was to report the potential for green synthesis of cerium oxide nanoparticles using plant extracts with a high content of phenolic metabolites.Methods: We have synthesized the CeO2 nano- and microparticles using Petroselinum crispum aqueous extract. The particles were characterized by UV-visible spectroscopy, IR spectroscopy, X-ray diffraction (XRD), scanning electron microscopy (SEM), transmission electron microscope (TEM) and dynamic light scattering analysis. For detection the reduction capacity of the extract the evaluation of total phenolic and flavonoid content as well as high-performance liquid chromatography-mass spectrometry (HPLC-MS) were performed. Biological activity of the particles was identified by bioluminescent tests and bio tests with Triticum vulgare.Results: Testing on T. vulgare showed that biogenic cerium dioxide powders stimulated the growth of up to 5-11,4% relative to intact samples, significantly increased the length of the leaves of seedlings and the root length. When seedings were exposed to the biosynthesized CeO2 particles, the level of chlorophylls was more stable and even slightly higher than control. Noticeable protective properties of the biosynthesized CeO2 powder under oxidation conditions were observed on the plant T. vulgare after a brief exposure (from 4 to 24 h).Conclusion: Thus, biogenic CeO2 can be potentially utilized in oxidative damage protection of agricultural plants.