RAPID CELL PURIFICATION SYSTEMS

• US 20140038171A1
• Filed: 03/07/2012
• Published: 02/06/2014
• Est. Priority Date: 03/07/2011
• Status: Active Grant

• Current Assignee: Accelerate Diagnostics, Inc.
• Original Assignee: Accelerated Technologies Corp.
• Inventors: Metzger, Steven W., Hance, Kenneth Robert
• Granted Patent: US 9,434,937 B2
• Time in Patent Office: Days
• US Class Current: 435/5
• CPC Class Codes: B03C 2201/18 Magnetic separation whereby the particles are suspended in a liquid
• CPC Class Codes: B03C 2201/26 for use in medical applications
• CPC Class Codes: B03C 5/005 Dielectrophoresis, i.e. dielectric particles migrating towards the region of highest field strength B03C5/02 takes precedence; electrophoresis B01D57/02
• CPC Class Codes: C12M 47/12 Purification C12M47/04 takes precedence
• CPC Class Codes: C12N 13/00 Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
• CPC Class Codes: C12N 2710/00051 Methods of production or purification of viral material
• CPC Class Codes: C12N 7/00 Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00
• CPC Class Codes: C12N 7/02 Recovery or purification
• CPC Class Codes: C12Q 1/02 involving viable microorganisms
• CPC Class Codes: C12Q 1/24 Methods of sampling, or inoculating or spreading a sample; Methods of physically isolating an intact microorganisms
• CPC Class Codes: G01N 1/34 Purifying; Cleaning processes or apparatus for extracting or separating nucleic acids from biological samples C12N15/1003
• CPC Class Codes: G01N 33/491 by separating the blood components G01N15/05 takes precedence
• CPC Class Codes: G02B 21/361 Optical details, e.g. image relay to the camera or image sensor G02B21/364 takes precedence; illumination details G02B21/06 and subgroups

Abstract

Methods and systems for purifying cells and/or viruses are provided. The sample is added to a well disposed in a medium. A potential is applied across the medium to cause the contaminants to enter one or more walls of the well, and retain the cells and/or viruses in the well. The cells and/or viruses can be removed from the well, and optionally adhered or fixed to a surface, or detected. In one embodiment, the cells and/or viruses may be retained in the well by embedding in the medium. The medium including the embedded cells and/or viruses may be excised or otherwise removed and transferred to a glass slide or other solid surface.

Claims

1. A method of purifying at least one of cells and viruses in a sample comprising:

adding the sample to a well disposed in a medium;

applying an electrical potential across the medium to cause contaminants to enter said medium through one or more walls of said well while retaining the at least one of the cells and/or viruses in said well; and

removing the at least one of the cells and viruses from said well.

2. The method of claim 1, wherein said cells are microorganisms.

8. The method of claim 1, wherein said medium comprises at least one of a filter or a hydrogel.

10. The method of claim 8, wherein said hydrogel comprises at least one of a polyacrylamide or agarose.

11. The method of claim 1, comprising placing a buffer in contact with said medium.

12. The method of claim 11, wherein said buffer comprises histidine and tris(hydroxymethyl)aminomethane.

13. The method of claim 1, further comprising immobilizing the at least one of the cells and viruses.

14. The method of claim 13, wherein the at least one the of cells and viruses are immobilized on a positively charged surface.

15. The method of any one of claim 1, further comprising detecting the at least one of the cells and viruses.

16. The method of claim 1, further comprising adding a chemical agent to the sample to increase a permeability of the medium to the contaminants.

17. The method of claim 1, further comprising applying a tangential flow to the medium to remove non-permeable contaminants from the surface of the medium.

18. The method of claim 1, wherein the electrical potential comprises an asymmetric alternating potential or a constant potential.

19. The method of claim 1, further comprising reversing the electrical potential to displace the at least one of the cells and viruses from the surface of the medium.

20. The method of claim 1, wherein removing the at least one of the cells and viruses from the well comprises removing at least a portion of the medium including a wall of the well having the at least one of the cells and viruses accumulated thereon.

21. The method of claim 20, further comprising fixing the portion of the medium for staining or extracting molecular samples for analysis.

22. The method of claim 1, further comprising providing a localization device, wherein the localization device draws the at least one of the cells and viruses to a discrete location in the well thereby localizing the at least one of the cells and viruses, and wherein the localization device is selected from at least one of a non-conductive material, a conductive material, and a discontinuous buffer system.

27. The method of claim 22, wherein the conductive material comprises at least two electrodes.

28. The method of claim 27, wherein the at least two electrodes comprises at least one sheet electrode.

30. The method of claim 13, wherein applying the electrical potential and reversing the electrical potential are performed iteratively.

31. The method of claim 1, wherein the electrical potential comprises a field polarity and the filed polarity is switched according to a programmed sequence.

3-7. -7. (canceled)

9. (canceled)

23-26. -26. (canceled)

29. (canceled)