Capture and elution of bio-analytes via beads that are used to disrupt specimens

• US 9,873,860 B2
• Filed: 01/12/2016
• Issued: 01/23/2018
• Est. Priority Date: 06/26/2009
• Status: Active Grant

• Current Assignee: Claremont Biosolutions, LLC
• Original Assignee: Claremont Biosolutions, LLC
• Inventors: Irvine, Bruce, Doebler, Robert W., Talbot, Ryan P.
• Primary Examiner(s): Brown, Melanie Yu
• Application Number:
  US14/993,953
• Publication Number:
  US 20160186127A1
• Time in Patent Office: 742 Days
• CPC Class Codes: C07H 1/06 Separation; Purification
• CPC Class Codes: C07K 1/22 Affinity chromatography or related techniques based upon selective absorption processes
• CPC Class Codes: C12N 1/066 by physical methods
• CPC Class Codes: G01N 2035/00524 Mixing by agitating sample carrier
• CPC Class Codes: G01N 33/54313 the carrier being characterised by its particulate form

Abstract

Lysing may include agitating a specimen in a chamber along with a medium that includes a particulate lysing material that has an affinity for a biological material. Lysing material may include beads or other material which may be coated that facilitates binding. The medium may include a fluid with a high salt or low pH level. The biological material may be eluted by lowering a concentration of salt or increasing a pH level. Lysing materials with two or more different affinities may be employed. Heating may be used. Lysing may be performed in a flow through apparatus.

Claims

1. A device to lyse cell specimens which include cells containing a biological material including at least one of nucleic acids or proteins or polypeptides, the device comprising:

an outer vessel that forms an outer chamber therein having a circular cross-section with an inner diameter D1, the outer vessel having at least two outer vessel ports to provide respective fluid communicative paths between the outer chamber and an exterior of the outer vessel;

an inner vessel that forms an inner chamber therein, the inner vessel having a circular cross-section with an outer diameter D2 sized to be closely and pivotably received in the outer chamber of the outer vessel with a fluid tight seal therebetween, the inner vessel having two or more inner vessel ports to provide fluid communicative paths between the inner chamber and an exterior of the inner vessel, the inner vessel ports positioned such that at a first rotational orientation of the inner vessel relative to the outer vessel, at least one of the inner vessel ports is aligned with one of the outer vessel ports and at a second rotational orientation of the inner vessel relative to the outer vessel, at least one inner vessel port is aligned with a different one of the outer vessel ports, wherein when aligned the inner vessel ports and the outer vessel ports provide a fluid communicative path between the exterior of the outer vessel and the inner chamber of the inner vessel;

an impeller sized to be received in the inner chamber of the inner vessel; and

a motor drivingly coupled to drive the impeller.

2. The device of claim 1 wherein in use the motor closes the inner chamber of the inner vessel from an exterior of the device.

3. The device of claim 1 wherein the inner chamber of the inner vessel has an inner diameter D3 and the motor has an outer diameter D4, at least a portion of the motor closely received by the inner chamber to provide a fluid tight seal therebetween.

4. The device of claim 3, further comprising:

at least one of a ring, a gasket or a washer which provides a fluid tight seal between the inner and the outer vessels or between the motor and the inner vessel.

5. The device of claim 1, further comprising:

at least one filter having a plurality of openings sized to block particulate lysing material.

6. The device of claim 1, further comprising:

a medium including a fluid and a particulate lysing material comprising a plurality of beads contained in the inner chamber of the inner vessel.

7. The device of claim 6 wherein particulate lysing material has a binding affinity for the biological material in the presence of the fluid and the fluid has a physical characteristic that induces binding of the biological material to the particulate lysing material, the binding affinity sufficient to bind at least some of the biological material to at least some of the particulate lysing material in the presence of the fluid.

8. The device of claim 6 wherein the particulate lysing material includes at least one of a plurality ceramic beads, a plurality of glass beads, a plurality of zirconium beads, a plurality of silica beads, a plurality of sand, or a plurality of beads with a metal core coated by latex.

9. The device of claim 6 wherein the particulate lysing material has diameters or smallest lateral dimensions in a range of 10 microns to 600 microns.

10. The device of claim 6 wherein the fluid has a high salt concentration of at least 500 millimolar.

11. The device of claim 6 wherein the fluid has a high salt concentration of at least 200 millimolar.

12. The device of claim 6 wherein the fluid has a pH from pH 3.5 to pH 5.

13. The device of claim 12 wherein the pH is no lower than 4.

14. A method to lyse cell specimens which include cells containing a biological material including at least one of nucleic acids or proteins or polypeptides using a device, the method comprising:

introducing the cell specimen to the device, the device comprising;

an outer vessel that forms an outer chamber therein having a circular cross-section with an inner diameter D1, the outer vessel having at least two outer vessel ports to provide respective fluid communicative paths between the outer chamber and an exterior of the outer vessel;

an inner vessel that forms an inner chamber therein which holds a particulate lysing material, the inner vessel having a circular cross-section with an outer diameter D2 sized to be closely and pivotably received in the outer chamber of the outer vessel with a fluid tight seal therebetween, the inner vessel having two or more inner vessel ports to provide fluid communicative paths between the inner chamber and an exterior of the inner vessel, the inner vessel ports positioned such that at a first rotational orientation of the inner vessel relative to the outer vessel, at least one of the inner vessel ports is aligned with one of the outer vessel ports and at a second rotational orientation of the inner vessel relative to the outer vessel, at least one inner vessel port is aligned with a different one of the outer vessel ports, wherein when aligned the inner vessel ports and the outer vessel ports provide a fluid communicative path between the exterior of the outer vessel and the inner chamber of the inner vessel;

an impeller sized to be received in the inner chamber of the inner vessel; and

a motor drivingly coupled to drive the impeller;

operating the motor to lyse the cell specimen;

rotating the inner vessel from a first rotational orientation to a second rotational orientation in which a one of the inner vessel ports aligns with a one of the outer vessel ports to provide a fluid communicative path between the inner chamber of the inner vessel and the exterior of the outer vessel; and

removing the biological material from the inner chamber of the inner vessel.

15. The method of claim 14, further comprising:

before introducing the cell specimen to the device, rotating the inner vessel to the first rotational orientation to align one of the inner vessel ports with a different one of the outer vessel ports than in the second rotational orientation.

16. The method of claim 14, further comprising:

introducing a fluid into the inner chamber of the inner vessel.

17. The method of claim 16 wherein introducing a fluid into the inner chamber of the inner vessel includes introducing a fluid in which the particulate lysing material has a binding affinity for the biological material in the presence of the fluid and the fluid has a physical characteristic that induces binding of the biological material to the particulate lysing material, the binding affinity sufficient to bind at least some of the biological material to at least some of the particulate lysing material in the presence of the fluid.

18. The method of claim 16 wherein introducing a fluid into the inner chamber of the inner vessel includes introducing the fluid into the particulate lysing material which includes at least one of a plurality of ceramic beads, a plurality of glass beads, a plurality of zirconium beads, a plurality of silica beads, a plurality of sand, or a plurality of beads with a metal core coated by latex.

19. The method of claim 16 wherein introducing a fluid into the inner chamber of the inner vessel includes introducing the fluid into the particulate lysing material which has diameters or smallest lateral dimensions in a range of 10 microns to 600 microns.

20. The method of claim 16 wherein introducing a fluid into the inner chamber of the inner vessel includes introducing the fluid having a high salt concentration of at least 500 millimolar.

21. The method of claim 16 wherein introducing a fluid into the inner chamber of the inner vessel includes introducing the fluid having a high salt concentration of at least 200 millimolar.

22. The method of claim 16 wherein introducing a fluid into the inner chamber of the inner vessel includes introducing the fluid having a pH from pH 3.5 to pH 5.

23. The method of claim 16 wherein introducing a fluid into the inner chamber of the inner vessel includes introducing the fluid having a pH no lower than 4.

24. The method of claim 14, further comprising:

flowing a volume of the cell specimen through the inner chamber that exceeds a volume of the inner chamber.

25. The method of claim 14, further comprising:

flowing the cell specimen through the inner chamber in a first direction at a first time and at a first rate, and in a second direction, opposite the first direction, at a second time and at a second rate.

26. The method of claim 14, further comprising:

before introducing a cell specimen to the device, placing the motor at least partially into the inner chamber of the inner vessel to seal the inner chamber from an exterior of the device.