Self-contained system, apparatus and method for continuous isolation of extracellular vesicles.
US-2024003897-A1 · Jan 4, 2024 · US
US9417165B2 · US · B2
| Field | Value |
|---|---|
| Publication number | US-9417165-B2 |
| Application number | US-201514805843-A |
| Country | US |
| Kind code | B2 |
| Filing date | Jul 22, 2015 |
| Priority date | Mar 19, 2013 |
| Publication date | Aug 16, 2016 |
| Grant date | Aug 16, 2016 |
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This disclosure is directed to a cap for obtaining a target analyte from a suspension. The cap introduces a magnetic field or a magnetic gradient to the tube to draw the target analyte bound to a particle to the cap. In one aspect, a cap includes a magnetic insert and a receiving piece. The magnetic insert includes a stopper and a magnet extending from the stopper; and, the receiving piece, which is configured to hold the magnetic insert, includes a receiving stopper and a sheath. The sheath may include imaging slides on opposite sides of the sheath. In another aspect, the cap may include a stopper and an embedded magnet. The cap may include an analysis piece on a bottom end of the stopper. In yet another aspect, the cap may include a fluid compartment and a filter at a bottom end of the stopper.
Opening claim text (preview).
We claim: 1. A system for separating a suspension suspected of containing a target analyte, comprising: a vessel having an open end to receive the suspension; a separating fluid having a density less than the density of the target analyte; and, a cap having a stopper including a bottom end and a top end, a magnet, and an analysis piece removably attached to the bottom end of the stopper, wherein the analysis piece is a cover slip or a slide, and wherein the bottom end is sized and shaped to fit within the open end of the vessel, wherein the target analyte is selected from the group consisting of a cell, a circulating tumor cell, a circulating endothelial cell, a fetal cell, a nucleated red blood cell, a red blood cell, a vesicle, a liposome, a protein, a nucleic acid, a naturally occurring microscopic unit having an enclosed membrane, an artificially prepared microscopic unit having an enclosed membrane, a parasite, a microorganism, a virus, and an inflammatory cell. 2. The system of claim 1 , wherein the vessel is a tube. 3. The system of claim 2 , further comprising a float sized to fit within the tube. 4. The system of claim 3 , further comprising a clamp to circumferentially apply pressure directed toward a central axis of the tube. 5. The system of claim 4 , wherein the clamp is selected from the group consisting of an O-ring, a pipe clamp, a hose clamp, a spring clamp, a strap clamp, a tie, a collet clamp and a piezoelectric ring. 6. The system of claim 1 , wherein the magnet is embedded within the stopper. 7. The system of claim 1 , wherein the analysis piece is attached to the bottom end of the stopper with a vacuum, an adhesive, or vacuum grease. 8. The system of claim 1 , wherein the bottom end of the stopper is angled. 9. The system of claim 1 , the cap further comprising at least one port to permit fluids to be introduced to and removed from the vessel. 10. The system of claim 1 , further comprising a solution containing at least one magnetic particle to conjugate with the target analyte to form a target analyte-particle complex thereby attracting the target analyte-particle complex to the cap. 11. The system of claim 10 , wherein the separating fluid has a surface tension that is not high enough to break the conjugation between the at least one magnetic particle and the target analyte. 12. The system of claim 11 , wherein the solution contains a plurality of magnetic particles, and wherein surface tension of the separating fluid is high enough to break a bond between at least one magnetic particle and a non-target analyte. 13. The system of claim 12 , wherein conjugation between the at least one magnetic particle and the target analyte is stronger than the bond between the at least one magnetic particle and the non-target analyte. 14. The system of claim 1 , further comprising a magnet external to the tube to draw the target analyte-particle complex through the separating fluid and closer to the cap. 15. A system for separating a suspension suspected of containing a target analyte, comprising: a vessel having an open end to receive the suspension; a separating fluid having a density less than the density of the target analyte; and, a cap having a stopper including a bottom end and a top end, a magnet, a sidewall extending outwardly from the bottom end to form a filter end, at least one filter including at least one pore in the filter end, and a fluid compartment located between the at least one filter and the bottom end, wherein the bottom end is sized and shaped to fit within the open end of the vessel, wherein the target analyte is selected from the group consisting of a cell, a circulating tumor cell, a circulating endothelial cell, a fetal cell, a nucleated red blood cell, a red blood cell, a vesicle, a liposome, a protein, a nucleic acid, a naturally occurring microscopic unit having an enclosed membrane, an artificially prepared microscopic unit having an enclosed membrane, a parasite, a microorganism, a virus, and an inflammatory cell. 16. A system for separating a suspension suspected of containing a target analyte, comprising: a vessel having an open end to receive the suspension; a separating fluid having a density less than the density of the target analyte; and, a cap having a stopper including a bottom end and a top end, a magnet, an inlet port to permit fluids to be introduced to the vessel, and an outlet port to permit fluids to be removed from the vessel, wherein the bottom end is sized and shaped to fit within the open end of the vessel, wherein the target analyte is selected from the group consisting of a cell, a circulating tumor cell, a circulating endothelial cell, a fetal cell, a nucleated red blood cell, a red blood cell, a vesicle, a liposome, a protein, a nucleic acid, a naturally occurring microscopic unit having an enclosed membrane, an artificially prepared microscopic unit having an enclosed membrane, a parasite, a microorganism, a virus, and an inflammatory cell. 17. A system for separating a suspension suspected of containing a target analyte, comprising: a vessel having an open end to receive the suspension; a separating fluid having a density less than the density of the target analyte; and, a cap having a magnetic insert having a stopper including a bottom end and a top end, and, a magnet extending outwardly from the bottom end; and a receiving piece having a receiving stopper including a bottom end and a top end; and, a sheath to receive the magnet, the sheath extending outwardly from the bottom end of the receiving stopper, the bottom end of the receiving stopper to fit within the opening of the vessel, the receiving stopper to receive the bottom end of the stopper of the magnetic insert, wherein the target analyte is selected from the group consisting of a cell, circulating tumor cell, a circulating endothelial cell, a fetal cell, a nucleated red blood cell, a red blood cell, a vesicle, a liposome, a protein, a nucleic acid, a naturally occurring microscopic unit having an enclosed membrane, an artificially prepared microscopic unit having an enclosed membrane, a parasite, a microorganism, a virus, and an inflammatory cell. 18. The system of claim 17 , the receiving piece having at least one slide removably attached to at least one side of the sheath.
sedimentation · CPC title
Preparing specimens for investigation {including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q}(mounting specimens on microscopic slides G02B21/34; means for supporting the objects or the materials to be analysed in electron microscopes H01J37/20 {; laboratory gas handling apparatus B01L5/00}) · CPC title
of liquid biological material · CPC title
Blood {(chemical methods for determining blood cell populations G01N33/5094; chemical analysis of blood groups or blood types G01N33/80)} · CPC title
Closing or opening means, corks, bungs · CPC title
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