Rapid cell purification systems

• US 9,434,937 B2
• Filed: 03/07/2012
• Issued: 09/06/2016
• Est. Priority Date: 03/07/2011
• Status: Active Grant

• Current Assignee: Accelerate Diagnostics, Inc.
• Original Assignee: Accelerate Diagnostics, Inc.
• Inventors: Metzger, Steven W., Hance, Kenneth Robert
• Primary Examiner(s): Lucas, Zachariah
• Assistant Examiner(s): Snyder, Stuart W
• Application Number:
  US14/004,145
• Publication Number:
  US 20140038171A1
• Time in Patent Office: 1,644 Days
• Field of Search: None
• US Class Current: 1/1
• 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:

1) adding a sample containing contaminants and at least one of cells and viruses to a well disposed in a porous medium;

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

3) reversing the electrical potential for a sufficient time to displace the at least one of the cells and viruses from the surface of the medium; and

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

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

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

4. The method of claim 3, wherein said hydrogel comprises at least one of polyacrylamide or agarose.

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

6. The method of claim 5, wherein said buffer comprises histidine and tris(hydroxymethyl)aminomethane.

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

8. 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.

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

10. 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.

12. The method of claim 1, wherein applying the electrical potential and reversing the electrical potential are performed iteratively.

14. The method of claim 1, further comprising mixing the sample.

15. The method of claim 14, wherein the sample is mixed by at least one of stirring the sample, repeatedly passing the sample through a pipette tip, and repeatedly forcing the sample through tubing connected to a syringe.

16. The method of claim 14, wherein the sample is mixed at least one of before application of the electrical potential, during application of the electrical potential, and after application of the electrical potential.

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

1) adding a sample containing contaminants and at least one of cells and viruses to a well disposed in a porous medium;

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

3) providing a localization device to said medium, wherein the localization device draws the at least one of the cells and viruses in said well to a discrete location in the well thereby localizing the at least one of the cells and viruses; and

4) removing the at least one of the cells and viruses from said well;

wherein the localization device is selected from at least one of a non-conductive material, a conductive material, and a discontinuous buffer system.

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

adding a sample containing contaminants and at least one of cells and viruses to a well disposed in a medium;

applying an electrical potential across the medium to cause the contaminants to enter said medium through a surface of one or more walls of said well while retaining the at least one of the cells and viruses in said well, wherein the electrical potential comprises a field polarity and the field polarity is switched according to a programmed sequence; and

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