Magnetosomes are biogenic magnetic nanoparticles with unique material properties, such as high crystallinity and strong magnetization as well as uniform and tunable shapes and sizes. In particular, the ability to genetically modify their enveloping membrane could render isolated magnetosomes as versatile tools for a variety of potential applications in the biomedical field. However, the biocompatibility of the bacterially produced particles has so far not been fully been assessed. In this study, different biocompatibility and toxicity factors of magnetosomes isolated from the model organism Magnetospirillum gryphiswaldense were investigated and tested with THP-1, Jurkat cells, and primary human-derived material such as peripheral blood mononuclear cells (PBMCs). The obtained data demonstrate generally good biocompatibility for all tested magnetosome concentrations as well as high blood compatibility. Only slight but not significant complement activation was observed, with no indications for plasma coagulation or hemolysis. However, the presence of lipopolysaccharides originating from the Gram-negative cell wall of M. gryphiswaldense caused activation of PBMCs expressed as significant cytokine release. Thus, this endotoxicity represents an issue for possible in vivo applications, which needs to be addressed in future studies. Nevertheless, the results show that, overall, magnetosomes have high potential for different (bio)medical applications and are already well-suited for the fields of in vitro diagnostics.

Bacterial magnetosomes ("MAGs") represent a promising class of magnetic iron oxide nanoparticles with exceptional material characteristics and high application potential in the biomedical and biotechnological field. For the surface functionalization of MAGs with different protein cargos, their enveloping membrane can be addressed by genetic means. However, the expression of foreign polypeptides as translational fusion to magnetosome membrane proteins is still laborious and lacks versatility as the generated particles are monospecific and thus restricted to predetermined functions. Utilizing the SpyTag-SpyCatcher (ST-SC) bioconjugate system, we here establish a flexible platform for the targeted nanoassembly of multifunctional MAGs that combines the rapidity of chemical coupling (e.g., by cross-linking reactions) and the unmatched selectivity and controllability of in vivo functionalization. MAGs genetically engineered to display either SC- or ST-connectors are shown to efficiently bind a variety of complementary tagged (protein) cargo. Specifically, we cover a broad spectrum of representative functional moieties and foreign cargo (such as enzymes, antibodies, fluorophores, and silica beads) with relevance in biotechnology and biomedicine and demonstrate the interchangeability of the MAGs-adapted ST-SC system. For the controlled generation of artificial shells surrounding the particles, SC-MAGs are effectively coated by protein-corona proteins. The potential of the here-provided toolkit is even more enhanced by using SC-MAGs as an affinity tool for selective protein pulldown in vitro and in vivo. Overall, this innovative technology turns bacterial MAGs into a flexible magnetic nanoscaffold for the targeted plug-and-play display of virtually unlimited additional functionalities, thereby generating a multitude of magnetic hybrid materials that can be used in many applications.

For the potential in vitro/in vivo applications of magnetic iron oxide nanoparticles, their stability in different physiological fluids has to be ensured. This important prerequisite includes the preservation of the particles' stability during the envisaged application and, consequently, their invariance with respect to the transfer from storage conditions to cell culture media or even bodily fluids. Here, we investigate the colloidal stabilities of commercial nanoparticles with different coatings as a model system for biogenic iron oxide nanoparticles (magnetosomes) isolated from magnetotactic bacteria. We demonstrate that the stability can be evaluated and quantified by determining the intensity-weighted average of the particle sizes (Z-value) obtained from dynamic light scattering experiments as a simple quality criterion, which can also be used as an indicator for protein corona formation.

Magnetosomes are magnetic nanoparticles biosynthesized by magnetotactic bacteria. Due to a genetically strictly controlled biomineralization process, the ensuing magnetosomes have been envisioned as agents for biomedical and clinical applications. In the present work, different stability parameters of magnetosomes isolated from Magnetospirillum gryphiswaldense upon storage in suspension (HEPES buffer, 4 °C, nitrogen atmosphere) for one year in the absence of antibiotics are examined. The magnetic potency, measured by the saturation magnetization of the particle suspension, drops to one-third of its starting value within this year-about ten times slower than at ambient air and room temperature. The particle size distribution, the integrity of the surrounding magnetosome membrane, the colloidal stability, and the biocompatibility turn out to be not severely affected by long-term storage.

Bacterial magnetosomes (MS) are well-defined membrane-enveloped single-domain iron oxide (magnetite) nanoparticles, which are susceptible to genetic and chemical engineering. Additionally, the possibility to manipulate these particles by external magnetic fields facilitates their application in biomedicine and biotechnology, e.g. as magnetic resonance imaging probes or for drug delivery purposes. However, current purification protocols are poorly characterized, thereby hampering standardized and reproducible magnetosome production and thus, reliable testing for in vivo applications. In that context, the establishment of reproducible particle isolation procedures as well as the identification of high quality control parameters and the evaluation of potential cytotoxic effects of purified particles are of major importance. In this study, we characterize a multi-step purification protocol for MS with regard to purity, iron content, size and polydispersity of magnetite particles. In addition, we address potential cytotoxic effects of isolated MS when incubated with mammalian cells. Overall, we provide a detailed overview of the process-structure relationship during the isolation of MS and thus, identify prerequisites for high-yield MS production and their future application in the biomedical and biotechnological field. STATEMENT OF SIGNIFICANCE: Magnetic nanoparticles are of increasing interest for a variety of biomedical and biotechnological applications. Due to their unprecedented material characteristics, bacterial magnetosomes represent a promising alternative to chemically synthesized iron oxide nanoparticles. As applications require well-defined, highly purified and fully characterized nanoparticles, reliable protocols are necessary for efficient and reproducible magnetosome isolation. In our study, we evaluate an improved magnetosome extraction procedure and monitor quality parameters such as particle size distribution, membrane integrity and purity of the suspension by a combination of physicochemical and biochemical methods. Furthermore, the cytotoxicity of the isolated magnetosomes is assessed using different cell lines. In summary, our study helps to establish prerequisites for many real-world applications of magnetosomes in the field of biotechnology and biomedicine.