CAPTURE AND ELUTION OF BIO-ANALYTES VIA BEADS THAT ARE USED TO DISRUPT SPECIMENS

• US 20100331522A1
• Filed: 06/24/2010
• Published: 12/30/2010
• Est. Priority Date: 06/26/2009
• Status: Active Grant

• Current Assignee: Claremont Biosolutions, LLC
• Original Assignee: Claremont Biosolutions, LLC
• Inventors: Doebler, Robert W., Niemz, Angelika Brigitte, Irvine, Bruce, Talbot, Ryan P., Erwin, Barbara
• Granted Patent: US 9,260,475 B2
• Time in Patent Office: Days
• US Class Current: 530/344
• 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 method of obtaining biological material, the method comprising:

introducing a specimen containing the biological material into a chamber; and

agitating the specimen in the chamber along with a medium that includes a particulate lysing material and a fluid to mechanically lyse the specimen, where the particulate lysing material has an affinity for the biological material in the presence of the fluid and the fluid has at least one physical characteristic that induces binding of the biological material to the particulate lysing material, the 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.

2. The method of claim 1 wherein the particulate lysing material comprises a plurality of beads and agitating the specimen in the chamber along with a medium that includes a particulate lysing material and a fluid to lyse the specimen includes agitating the specimen in the chamber along with the plurality of beads.

3. The method of claim 1 wherein the particulate lysing material 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 a material that facilitates binding of the biological material and agitating the specimen in the chamber along with a medium that includes a particulate lysing material and a fluid to lyse the specimen includes agitating the specimen in the chamber along with the at least one of the plurality of ceramic beads, the plurality of glass beads, the plurality of zirconium beads, the plurality of silica beads, or the plurality of sand.

4. The method of claim 1 wherein agitating the specimen in the chamber along with a medium that includes a particulate lysing material and a fluid to lyse the specimen includes agitating the specimen in the chamber along with particulate lysing material having diameters or smallest lateral dimensions in the range of approximately 10 microns to approximately 600 microns.

5. The method of claim 1 wherein the fluid has a high salt concentration, and agitating the specimen in the chamber along with a medium that includes a particulate lysing material and a fluid to lyse the specimen includes agitating the specimen in the chamber with the fluid having the high salt concentration.

6. The method of claim 5 wherein agitating the specimen in the chamber with the fluid having the high salt concentration includes agitating the specimen in chamber with the fluid having the salt concentration of at least about 200 millimolar.

7. The method of claim 1 wherein the fluid has a low pH, and agitating the specimen in the chamber along with a medium that includes a particulate lysing material and a fluid to lyse the specimen includes agitating the specimen in the chamber with the fluid having the low pH.

8. The method of claim 7 wherein agitating the specimen in the chamber with the fluid having the low pH includes agitating the specimen in the chamber with the fluid having the pH no lower than about 4.

9. The method of claim 1 wherein the medium in the chamber is free of any chemical lysing agents.

10. The method of claim 1 wherein agitating the specimen in the chamber along with a medium that includes a particulate lysing material and a fluid to lyse the specimen includes oscillating an arm on which the chamber is mounted.

11. The method of claim 1 wherein agitating the specimen in the chamber along with a medium that includes a particulate lysing material and a fluid to lyse the specimen includes driving an impeller having a number of blades located at least partially in the chamber.

12. The method of claim 1, further comprising:

eluting the biological material from at least the particulate lysing material.

13. The method of claim 12 wherein eluting the biological material from at least the particulate lysing material includes at least one of lowering a salt concentration or increasing a pH of an environment in which the particulate lysing material is contained.

14. The method of claim 13 wherein lowering a salt concentration or increasing a pH of an environment in which the particulate lysing material is contained includes lowering a salt concentration or increasing a pH of the fluid in the chamber.

15. The method of claim 12 wherein eluting the biological material from at least the particulate lysing material includes eluting the material without any washing.

16. The method of claim 12 wherein eluting the biological material from at least the particulate lysing material includes eluting at least one of DNA material, a protein or polypeptide material, or another cell component material.

17. The method of claim 12, further comprising:

heating the biological material after eluting the biological material from at least the particulate lysing material.

18. The method of claim 1, further comprising:

heating the biological material after lysing the specimen.

19. The method of claim 1, further comprising:

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

20. The method of claim 1, further comprising:

flowing the specimen through the 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.

21. The method of claim 1 wherein the medium includes two particulate lysing materials that have been modified to bind biological materials having two different chemistries, a first particulate lysing material having been modified to bind biological material having a first chemistry and a second particulate lysing material having been modified to bind biological material having a second chemistry.

22. The method of claim 21 wherein the first particulate lysing material having been modified to bind biological material having a first chemistry is a particulate lysing material having been modified to bind DNA and the second particulate lysing material having been modified to bind biological material having a second chemistry is a particulate lysing material having been modified to bind protein or polypeptide.

23. The method of claim 22 further comprising:

labeling the protein or polypeptide with a solution-phase tagged ligand during binding to facilitate subsequent detection of the protein or polypeptide.

24. The method of claim 21, further comprising:

separately eluting biological material having a first chemistry and biological material having a second chemistry.

25. The method of claim 24 wherein the biological material having a first chemistry is DNA and the biological material having a second chemistry is protein or polypeptide.

26. The method of claim 25 wherein the DNA is eluted with low salt and the protein or polypeptide is eluted with urea.

27. The method of claim 1, further comprising:

separately transferring unbound biological material to a separate chamber or container for analysis or further processing.

28. A system to lyse specimens and isolate specific biological materials from the lysed specimens, the system comprising:

a chamber to receive a specimen containing a biological material to be isolated therefrom;

a medium that includes a particulate lysing material and a fluid, the particulate lysing material having an affinity for the biological material in the presence of the fluid and the fluid having a physical characteristic that induces binding of the biological material to the particulate lysing material, the 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; and

an agitator selectively operable to agitate the specimen in the chamber along with the medium that includes the particulate lysing material and the fluid to mechanically lyse the specimen and concurrently bind at least some of the biological material to at least some of the particulate lysing material.

29. The system of claim 28 wherein the particulate lysing material includes a plurality of beads.

30. The system of claim 28 wherein the particulate lysing material includes at least one of a plurality 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 a material that facilitates binding of the biological material.

31. The system of claim 28 wherein the particulate lysing material has diameters or smallest lateral dimensions in the range of approximately 10 microns to approximately 600 microns.

32. The system of claim 28 wherein the fluid has a high salt concentration.

33. The system of claim 32 wherein the high salt concentration is at least about 200 millimolar.

34. The system of claim 28 wherein the fluid has a low pH.

35. The system of claim 34 wherein the pH no lower than about 4.

36. The system of claim 28 wherein the medium in the chamber is free of any chemical lysing agents.

37. The system of claim 28 wherein the agitator includes at least one motor and an arm that holds the chamber, the arm coupled to the motor to be oscillatingly driven thereby.

38. The system of claim 28 wherein the agitator includes at least one motor and an impeller that is positionable at least partially in the chamber, the impeller coupled to the motor to be oscillatingly driven thereby.

39. The system of claim 28 wherein the biological material includes at least one of DNA material, a protein or polypeptide material, or another cell component material.

40. The system of claim 28, further comprising:

a pump operable to flow a volume of the specimen through the chamber that exceeds a volume of the chamber.

41. The system of claim 28, further comprising:

a pump operable to flow the specimen through the 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.

42. The system of claim 28, further comprising:

a heater in thermal contact with a pump, the heater selectively operable to heat the biological material.

43. The system of claim 28 wherein the medium includes particulate lysing materials that have been modified to bind biological materials of two different chemistries, a first particulate lysing material having been modified to bind biological material having a first chemistry and a second particulate lysing material having been modified to bind biological material having a second chemistry.

44. The system of claim 43 wherein the first particulate lysing material having been modified to bind biological material having a first chemistry is a particulate lysing material having been modified to bind DNA and the second particulate lysing material having been modified to bind biological material having a second chemistry is a particulate lysing material having been modified to bind protein or polypeptide.

45. The system of claim 28, further comprising:

a chamber or container for receipt of unbound biological material.