Targeted magnetic separation of biomolecules and cells using earthicle-based ferrofluids

Abstract

Targeting specific molecular or cell populations within single tissues or multicomponent in vitro systems is a most sought goal in biomedicine. Here we report on targeted magnetic separation of cells and biomolecules using a ferrofluid comprising superparamagnetic iron-oxide/silicate/carbon core/shell/crust nanoparticles in combination with a handheld, 2.5 cm³ NdFeB magnet (≤180 mT) and one minute exposure time. Ferrofluids were highly effective at separating (i) biomolecules, (ii) bacteria and (iii) eukaryotic cells from solutions, and they also exhibited selectivity in the separation of all three families of entities. Specifically, they were more effective at separating the negatively charged protein, albumin in the presence of the external magnetic field, but were more effective at precipitating the positively charged protein, lysozyme without the application of the external field. Because of the more effective sorption of proteins than carbohydrates on carbon and the shielding of peptidoglycans by the transmembrane proteins and hydrophilic heads of the outer membrane amphiphiles in Gram-negative bacteria, they were separated more effectively than their Gram-positive counterparts. Ferrofluids were also more efficient at separating the clinical isolate, methicillin-resistant version of S. aureus (MRSA) than its regular, lab strain and the effect is thought to be due to structural changes to the cell envelope caused by the overexpression of efflux pumps or by the higher rate of conjugation conditioning horizontal gene transfer in MRSA than in the regular, nonresistant strain. Ferrofluids also displayed a greater affinity for the cancer cells than for the normal, primary cells and allowed for targeted separation of the former after the cells were allowed to uptake the nanoparticles for 24 h. This selectivity should allow for an effective separation of cancer cells interspersed within a healthy cell population. Interaction with bacterial and eukaryotic cells was driven neither by electrostatic attraction nor chemisorption, but by weaker, van der Waals and π-interactions. Adsorption was also endothermic, irreversible for the most part, and more favorable at high concentrations, as inferred by comparison with Langmuir, Freundlich, Temkin and Dubinin-Radushkevich isotherms. These targeted effects are relevant for numerous fields of biomedicine and biotechnologies and require further insight for optimization and translation.