Anionic Polyethylene Glycol Research Articles

Insight into the molecular mechanism behind PEG-mediated stabilization of biofluid lipases

Bioconjugates established between anionic polyethylene glycol (PEG) based polymers and cationic proteins have proven to be a promising strategy to engineer thermostable biocatalysts. However, the enzyme activity of these bioconjugates is very low and the mechanism of non-covalent PEG-stabilization is yet to be understood. This work presents experimental and molecular dynamics simulation studies, using lipase-polymer surfactant nanoconjugates from mesophile Rhizomucor miehei (RML), performed to evaluate the effect of PEG on enzyme stability and activity. Results demonstrated that the number of hydrogen bonds between the cationized RML and PEG chain correlates with enzyme thermostability. In addition, an increase of both the number of PEG-polymers units and cationization degree of the enzyme leads to a decrease of enzyme activity. Modelling with SAXS data of aqueous solutions of the biofluid lipases agrees with previous hypothesis that these enzymes contain a core constituted of folded protein confined by a shell of surfactants. Together results provide valuable insight into the mechanism of non-covalent PEG mediated protein stabilization relevant for engineering active and thermostable biofluids. Furthermore, the first biofluids RML with activity comparable to their cationized counterpart are presented.

Tuning the Multifunctionality of Iron Oxide Nanoparticles Using Self-Assembled Mixed Lipid Layers.

We present an approach to tuning the multifunctionality of iron oxide nanoparticles (IONs) using mixed self-assembled monolayers of cationic lipid and anionic polyethylene glycol (PEG) lipid. By forming stable, monodispersed lipid-coated IONs (L-IONs) through a solvent-exchange technique, we were able to demonstrate the relationship between surface charge, the magnetic transverse relaxivity (r2 from T2-weighted images), and the binding capacity of small interfering ribonucleic acids (siRNAs) as a function of the cationic-to-anionic (PEG) lipid ratio. These properties were controlled by the cationic charge and the PEG conformation; relaxivity and siRNA binding could be varied in the mushroom and brush regimes but not at high brush densities. In vitro results combining cell viability, uptake, and transfection efficiency using HeLa cells suggest that the functional physicochemical and biological properties of L-IONs may be best achieved using catanionic lipid coatings near equimolar ratios of cationic to anionic PEG-lipids.

Effects of surface charge and ligand type of Au nanoparticles on green fluorescent protein-expressing Escherichia coli

The effects of the surface charge and ligand type of three types of Au nanoparticles (NPs), namely anionic polyethylene glycol (PEG)-Au NPs, anionic citrate (Cit)-Au NPs, and cationic branched polyethylenimine (bPEI)-Au NPs, on green fluorescent protein (GFP)-expressing Escherichia coli were evaluated through the combined analysis of optical density (OD) and fluorescence intensity (FI). OD and FI can provide information about cell growth and metabolism of bacteria, respectively. The results demonstrated that PEG- and Cit-Au NPs had no major effects on the OD and FI of GFP-expressing bacteria. However, it was found that Cit-Au NPs may slightly influence cell metabolism at higher concentrations, although it is necessary to perform further in-depth study to clarify this issue. Cationic bPEI-Au NPs showed significant effects on cell density and metabolism of E. coli, with an especially strong effect on metabolism. The combined analysis of OD and FI may be useful for monitoring the effects of a wide range of nanomaterials on microorganisms.

475. Anionic Polyethyleneglycol Lipids Added to Cationic Lipoplexes Increase Their Plasmatic Circulation Time

Cationic liposomes have been widely sensed as good DNA compacting delivery agents. Although their use generally met with encouraging results in vitro, the results in vivo were rather disappointing, as they strongly interact with the blood components before they can reach the therapeutic target. Polyethyleneglycol (PEG) shielding has been proposed as a way to alleviate this effect, but was still found unsatisfactory in most instances for systemic administration. We demonstrate here that the insertion of anionic functions between the lipid part and the PEG, at a correct distance to favor electrostatic interactions with the outer cationic layer of the lipoplexes, provides not only a decrease in the mean peripheral charge of the lipoplex ( z potential), but also a greater colloidal stability of the particles in the presence of serum. Transfection in the lung is also decreased with negatively charged PEG shielding, although no significant changes are observed in the tumor. This encouraging new approach should consequently be combined with active extra-cellular receptor targeting to achieve the desired delivery of the therapeutic DNA to tumor tissues.