Isolating a target analyte from a body fluid
AssigneeDNAE Group Holdings Limited
InventorsLisa-Jo Ann Clarizia, Eddie W. Adams, Sergey A. Dryga
Life of this patent
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Abstract
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.
Claims9
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.
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.
Dependencies are inferred from the claim text.