Superparamagnetic Nanoparticle Delivery of DNA Vaccine

Abstract

The efficiency of delivery of DNA vaccines is often relatively low compared to protein vaccines. The use of superparamagnetic iron oxide nanoparticles (SPIONs) to deliver genes via magnetofection shows promise in improving the efficiency of gene delivery both in vitro and in vivo. In particular, the duration for gene transfection especially for in vitro application can be significantly reduced by magnetofection compared to the time required to achieve high gene transfection with standard protocols. SPIONs that have been rendered stable in physiological conditions can be used as both therapeutic and diagnostic agents due to their unique magnetic characteristics. Valuable features of iron oxide nanoparticles in bioapplications include a tight control over their size distribution, magnetic properties of these particles, and the ability to carry particular biomolecules to specific targets. The internalization and half-life of the particles within the body depend upon the method of synthesis. Numerous synthesis methods have been used to produce magnetic nanoparticles for bioapplications with different sizes and surface charges. The most common method for synthesizing nanometer-sized magnetite Fe3O4 particles in solution is by chemical coprecipitation of iron salts. The coprecipitation method is an effective technique for preparing a stable aqueous dispersions of iron oxide nanoparticles. We describe the production of Fe3O4-based SPIONs with high magnetization values (70 emu/g) under 15 kOe of the applied magnetic field at room temperature, with 0.01 emu/g remanence via a coprecipitation method in the presence of trisodium citrate as a stabilizer. Naked SPIONs often lack sufficient stability, hydrophilicity, and the capacity to be functionalized. In order to overcome these limitations, polycationic polymer was anchored on the surface of freshly prepared SPIONs by a direct electrostatic attraction between the negatively charged SPIONs (due to the presence of carboxylic groups) and the positively charged polymer. Polyethylenimine was chosen to modify the surface of SPIONs to assist the delivery of plasmid DNA into mammalian cells due to the polymer's extensive buffering capacity through the "proton sponge" effect.

Unexpected magnetic attraction: Evidence for an organized energy field in the human body

“…Multiple studies since 2007 describe research with highly magnetic nanoparticles thus fueling allegations that the COVID-19 vaccine was imbedded with these substances [5][6][7][8][9][10][11][12][13][14][15][16][17]. Theories ascribing magnet attachment to highly magnetic nanoparticles embedded in the COVID-19 vaccine are clearly refuted by this study and could not be rationally challenged.…”

Nanoparticle Systems for Cancer Vaccine

“…To address these issues, modification with other components such as polycationic polymer can be used to provide the surface with sufficient capacity for functionalization and antigen binding. Superparamagnetic iron oxide nanoparticles (SPIONs) modified with cationic polyethyleneimine (PEI) polymer are promising DNA vaccine vectors due to their high buffering capacity [106]. Magnetic fields offer the capacity of enhanced cellular uptake and DC maturation.…”

Polymer- coated magnetic nanoparticles formulation enhanced blood-stage Malaria DNA vaccine delivery with the best PEI/Fe ratio

“…Synthesis and characterization of the Polyethylenimine-coated magnetic nanoparticles is described in detail in our previous paper (Al-Deen et al, 2013, Al-Deen et al, 2014b. Two concentrations of iron oxide suspension were examined in this study (0.1 and 0.5 mg/ml).…”

“…VR1020-PyMSP119 and VR1020-GFP plasmids were amplified using Escherichia coli DH5α as it is explained in detail in our previous papers (Al-Deen et al, 2014b. A single colony of E. coli harbouring plasmid VR1020-PyMSP119 and VR1020-GFP were picked out from a freshly streaked selective plate and inoculated in a 10 ml starter culture medium (LB broth containing 10 g NaCl, 10 g Bacto Tryptone, 5 g yeast, and 100 μg/ ml of Kanamycin) for 8 h at 37 •C with vigorous shaking at 200 rpm.…”