Devices and methods for enrichment and alteration of cells and other particles

• US 20070196820A1
• Filed: 09/15/2005
• Published: 08/23/2007
• Est. Priority Date: 04/05/2005
• Status: Abandoned Application

• Current Assignee: Gpb Scientific, LLC, Mehmet Toner
• Original Assignee: Mehmet Toner
• Inventors: Toner, Mehmet, Kapur, Ravi
• Application Number:
  US11/227,904
• Publication Number:
  US 20070196820A1
• Time in Patent Office: Days
• US Class Current: 435/5
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 2400/0409 centrifugal forces
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 2400/043 magnetic forces
• CPC Class Codes: B01L 2400/0472 Diffusion
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/086 using baffles or other fixed flow obstructions
• CPC Class Codes: B01L 3/502746 characterised by the means for controlling flow resistance, e.g. flow controllers, baffles B01L3/502738 takes precedence
• CPC Class Codes: B01L 3/502753 characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation separation in general B01D; microapparatus for analysis using electrophoresis G01N27/44791; sample preparation G01N1/28
• CPC Class Codes: B01L 3/502761 specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules investigating characteristics of particles G01N15/00
• CPC Class Codes: B03C 1/30 Combinations with other devices, not otherwise provided for
• CPC Class Codes: B03C 1/32 acting on the medium containing the substance being separated, e.g. magneto-gravimetric-, magnetohydrostatic-, or magnetohydrodynamic separation
• CPC Class Codes: B03C 2201/18 Magnetic separation whereby the particles are suspended in a liquid
• CPC Class Codes: B33Y 80/00 Products made by additive manufacturing
• CPC Class Codes: C12M 47/04 Cell isolation or sorting purging biological preparations of unwanted cells C12N5/0081, determining the presence or kind of microorganism C12Q1/04
• CPC Class Codes: C12M 47/06 Hydrolysis; Cell lysis; Extraction of intracellular or cell wall material lysis of microorganisms C12N1/06; extracting or separating nucleic acids from biological samples C12N15/1003
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: G01N 1/40 : Concentrating samples
• CPC Class Codes: G01N 1/4077 : by other techniques involving separation of suspended solids
• CPC Class Codes: G01N 2035/00237 : Handling microquantities of analyte, e.g. microvalves, capillary networks
• CPC Class Codes: G01N 33/5044 : involving specific cell types
• CPC Class Codes: Y10T 137/0318 : Processes
• CPC Class Codes: Y10T 137/8593 : Systems
• CPC Class Codes: Y10T 436/25 : including sample preparation
• CPC Class Codes: Y10T 436/25375 : Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]

Abstract

The invention features a device for the deterministic separation of analytes coupled to a reservoir containing a reagent that alters a magnetic propert of the analyte. Exemplary methods include the enrichment of a sample in a desired analyte (e.g., using deterministic separation) or the alteration of a desired analyte in the device. The devices and methods may be advantageously employed to enrich for rare cells, e.g., fetal cells or epithelial cells, present in a sample, e.g., maternal blood.

Claims

1. A device for producing a sample enriched in an analyte, said device comprising:

(a) a first channel comprising a structure that deterministically deflects particles having a hydrodynamic size above a critical size in a direction not parallel to the average direction of flow in said structure, wherein said particles are analyte particles or are a non-analyte component of said sample; and

(b) a reservoir fluidly coupled to an output of said first channel through which said analyte passes into said reservoir, wherein said reservoir comprises a reagent that alters a magnetic property of said analyte.

2. The device of claim 1, wherein said first channel is a microfluidic channel.

3. The device of claim 1, wherein said structure comprises an array of obstacles that form a network of gaps, wherein a fluid passing through said gaps is divided unequally into a major flux and a minor flux so that the average direction of the major flux is not parallel to the average direction of fluidic flow in said channel.

4. The device of claim 3, wherein said array of obstacles comprises first and second rows, wherein the second row is displaced laterally relative to the first row so that fluid passing through a gap in the first row is divided unequally into two gaps in the second row.

5. The device of claim 1, wherein said analyte has a hydrodynamic size greater than said critical size.

6. The device of claim 1, wherein said analyte has a hydrodynamic size smaller than said critical size.

7. The device of claim 1, further comprising a magnetic force generator capable of generating a magnetic field.

8. The device of claim 7, wherein said magnetic force generator comprises a region of magnetic obstacles disposed in a second channel.

9. The device of claim 8, wherein at least a portion of said magnetic obstacles comprise a permanent magnet.

10. The device of claim 8, wherein at least a portion of said magnetic obstacles comprise a non-permanent magnet.

11. The device of claim 8, wherein said obstacles are ordered in a two-dimensional array.

12. The device of claim 8, wherein said second channel is a microfluidic channel.

13. The device of claim 1, wherein said reservoir further comprises a second channel comprising a magnet.

14. The device of claim 1, wherein said reagent alters an intrinsic magnetic property of said one or more analytes.

15. The device of claim 14, wherein said reagent comprises sodium nitrite.

16. The device of claim 1, wherein said reagent binds to said one or more analytes.

17. The device of claim 16, wherein said reagent comprises a magnetic particle.

18. The device of claim 17, wherein said magnetic particle comprises an antibody or an antigen-binding fragment thereof.

19. The device of claim 18, wherein said antibody is anti-CD71, anti-CD36, anti-CD45, anti-GPA, anti-antigen i, anti-CD34, or anti-fetal hemoglobin.

20. The device of claim 16, wherein said reagent comprises holo-transferrin.

21. A method for producing a sample enriched in a first analyte relative to a second analyte, said method comprising:

(a) applying at least a portion of said sample to a device comprising a structure that deterministically deflects particles having a hydrodynamic size above a critical size in a direction not parallel to the average direction of flow in said structure, thereby producing a second sample enriched in said first analyte and comprising said second analyte;

(b) combining said second sample with a reagent that alters a magnetic property of said first analyte to produce an altered first analyte; and

(c) applying a magnetic field to said second sample, wherein said magnetic field generates a differential force to physically separate said altered first analyte from said second analyte, thereby producing a sample enriched in said first analyte.

22. The method of claim 21, wherein said reagent binds to said first analyte.

23. The method of claim 21, wherein said reagent alters an intrinsic magnetic property of said first analyte.

24. The method of claim 23, wherein said reagent comprises sodium nitrite.

25. The method of claim 21, wherein said reagent comprises a magnetic particle that binds to or is incorporated into said first analyte.

26. The method of claim 25, wherein said magnetic particle comprises an antibody or an antigen-binding fragment thereof.

27. The method of claim 26, wherein said antibody is anti-CD71, anti-GPA, anti-antigen i, anti-CD45, anti-CD34, or anti-fetal hemoglobin.

28. The method of claim 21, wherein said analyte has a hydrodynamic size greater than said critical size.

29. The method of claim 21, wherein said analyte has a hydrodynamic size smaller than said critical size.

30. The method of claim 21, wherein said sample comprises a maternal blood sample.

31. The method of claim 21, wherein said first analyte is a cell, an organelle, or a virus.

32. The method of claim 31, wherein said cell is a bacterial cell, a fetal cell, or a blood cell.

33. The method of claim 32, wherein said blood cell is a fetal red blood cell.

34. The method of claim 31, wherein said organelle is a nucleus.

35. A method of producing a sample enriched in red blood cells relative to a second blood component, said method comprising:

(a) contacting a sample comprising red blood cells with a reagent that oxidizes iron to produce oxidized hemoglobin; and

(b) applying a magnetic field to said sample, wherein said red blood cells having oxidized hemoglobin are attracted to said magnetic field to a greater extent than said second blood component, thereby producing said sample enriched in said red blood cells.

36. The method of claim 35, wherein said red blood cells are fetal red blood cells.

37. The method of claim 36, wherein said second blood component is a maternal blood cell.

38. The method of claim 35, wherein prior to said step (a), said sample is enriched for said red blood cells.

39. The method of claim 38, wherein said enriching is performed by applying at least a portion of said sample to a device comprising a structure that deterministically deflects particles having a hydrodynamic size above a critical size in a direction not parallel to the average direction of flow in said structure.

40. The method of claim 39, wherein fetal red blood cells are enriched relative to maternal red blood cells.

41. A device for producing a sample enriched in red blood cells, said device comprising:

(a) an analytical device that enriches said red blood cells based on size, shape, deformability, or affinity; and

(b) a reservoir comprising a reagent that oxidizes iron, wherein said reagent increases the magnetic responsiveness of said red blood cells.

42. The device of claim 41, wherein said analytical device comprises a first channel comprising a structure that deterministically deflects particles having a hydrodynamic size above a critical size in a direction not parallel to the average direction of flow in said structure.

43. The device of claim 41, wherein said reagent is sodium nitrite.