Isolating a target analyte from a body fluid
The invention generally relates to using magnetic particles and magnets to isolate a target analyte from a body fluid sample. In certain embodiments, methods of the invention involve introducing magnetic particles including a target-specific binding moiety to a body fluid sample in order to create a mixture, incubating the mixture to allow the particles to bind to a target, applying a magnetic field to capture target/magnetic particle complexes on a surface, and washing with a wash solution that reduces particle aggregation, thereby isolating target/magnetic particle complexes.
1. A method for isolating pathogen in a biological sample, the method comprising:
exposing a body fluid sample comprising blood cells and pathogen to a plurality of magnetic particles, the particles comprising at least about 70% magnetic material by weight and being functionalized for binding pathogen;
preventing lysis of the blood cells in the sample by mixing the particles and the sample with a buffer that substantially prevents lysis of blood cells, reduces particle aggregation, and comprises about 75 mM Tris(hydroxymethyl)-aminomethane hydrochloride, thereby forming complexes between the pathogen and magnetic particles, said buffer further comprising about 300 mM NaCl, and about 0.1% polysorbate 20;
flowing the complexes through a flow-through capture cell having a surface upon which a magnetic field is applied; and
isolating complexed pathogen by applying the magnetic field to the flow-through capture cell,
wherein the mixing step is performed prior to the flowing step.
2. The method of claim 1 , wherein the particles are coated with one or more antibodies capable of binding at least one pathogen.
3. The method of claim 1 , wherein the pathogen is selected from the group consisting of bacteria and viruses.
4. The method of claim 1 , wherein the body fluid sample is blood.
5. A method for isolating pathogen in a biological sample, the method comprising:
diluting a body fluid sample comprising blood cells and pathogen while preventing lysis of the blood cells by mixing the sample at a ratio of about 1:1 with a buffer that substantially prevents lysis of blood cells, reduces particle aggregation, and comprises about 75 mM Tris(hydroxymethyl)-aminomethane hydrochloride, about 300 mM NaCl, and about 0.1% polysorbate 20;
exposing the diluted sample to magnetic nanoparticles, the nanoparticles comprising at least about 70% magnetic material by weight and being functionalized for binding pathogen;
incubating the diluted sample and nanoparticles, thereby forming complexes between pathogen and the magnetic nanoparticles;
flowing the complexes through a flow-through capture cell having a surface upon which a magnetic field is applied; and
isolating the complexed pathogen by applying the magnetic field to the flow-through capture cell,
wherein the mixing step is performed prior to the flowing step.
6. The method of claim 5 , wherein the nanoparticles are functionalized with antibody that specifically binds to pathogen.
7. The method of claim 6 , wherein the pathogen is a bacterium.
8. The method of claim 5 , wherein osmolality of the sample is substantially maintained in the diluting step.
9. The method of claim 5 , wherein the incubating step is performed for from about 10 seconds to about 2 hours.