Stable nanomagnetic particle dispersions
Processes and compositions are described for preparing new, colloidally stable, coated nanomagnetic particles useful for both in-vitro and in-vivo biomedical applications, including cell targeting and capturing cells, microorganisms, and cellular organelles or entities such as exosomes. These nanomagnetic particles can also be used as imaging contrast agents due to their small size and high magnetic moment. The nanomagnetic particles include a series of sequentially added, stabilizing surface coatings rendered onto nano-sized magnetic crystal clusters (e.g., magnetite particles) to impart colloidal stability in complex biological samples with minimal leaching of the coating materials, high binding capacity, and low non-specific binding. Another benefit of this invention is the ability to utilize both external and internal magnetic field-generating separation devices to effect separation of the magnetic nanoparticles.
1. A method of making targeted nanomagnetic particles, the method comprising:
(a) obtaining magnetic core particles having a plurality of diameters ranging from about 30 nm to about 500 nm;
(b) encapsulating the magnetic core particles with a glass layer to form first encapsulated magnetic core particles, wherein the glass layer optionally has a thickness of about 1 nm to about 50 nm;
(c) encapsulating the first encapsulated magnetic core particles in a composite layer comprising a mixture of a protein and polymer to form second encapsulated magnetic core particles, wherein the composite layer of protein, which is bovine albumin or human albumin and polymer, which is dextran, optionally has a thickness of about 5 nm to about 50 nm, and
(d) binding, covalently, targeting moieties to the protein and polymer composite layer of the second encapsulated magnetic core particles, wherein the targeting moieties comprise one member of a bioaffinity ligand pair, optionally wherein the targeting moieties are independently selected from the group consisting of antibodies, an antigen-binding antibody fragments, a cell surface receptor, ligand-binding extracellular domains of a cell surface receptor, nucleic acids, avidin, streptavidin, biotin, pharmaceutical compounds for purposes of targeted drug delivery; and, optionally,
(e) binding, optionally covalently, a pharmaceutical compound or detectable label to the second encapsulated magnetic core particles.
2. A method according to claim 1 wherein the magnetic core particles are peptized.
3. A method of forming biomolecule/particle complexes, comprising binding targeted nanomagnetic particles to biomolecules of interest that are directly or indirectly specifically reactive with the targeting moieties of the targeted nanomagnetic particles to form biomolecule/particle complexes, wherein the targeted nanomagnetic particles have a plurality of diameters ranging from about 30 nm to about 500 nm, each nanomagnetic particle comprising:
(a) a magnetic core particle;
(b) a glass layer encapsulating the magnetic core particle;
(c) a protein and polymer composite layer bound to and encapsulating the glass layer; and
(d) a targeting moiety that comprises one member of a bioaffinity ligand pair bound to the protein and polymer composite layer, wherein the bioaffinity ligand is capable of specifically binding to the biomolecule of interest, wherein the protein and polymer composite layer comprises a mixture of a protein and polymer, wherein the protein is bovine albumin or human albumin and polymer is dextran.
4. A method according to claim 3 wherein the binding occurs in vitro or in vivo.