System for size based separation and analysis

• US 20070059781A1
• Filed: 09/15/2005
• Published: 03/15/2007
• Est. Priority Date: 09/15/2005
• Status: Abandoned Application

• Current Assignee: Gpb Scientific, LLC, Mehmet Toner
• Original Assignee: Mehmet Toner
• Inventors: Kapur, Ravi, Gray, Darren, Barber, Tom, Carvalho, Bruce, Toner, Mehmet, Huang, Lotion
• Application Number:
  US11/229,336
• Publication Number:
  US 20070059781A1
• Time in Patent Office: Days
• US Class Current: 435/7.210
• 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 2400/0409 centrifugal forces
• CPC Class Codes: B01L 2400/0472 Diffusion
• 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: B82Y 15/00 Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: G01N 1/40 Concentrating samples
• CPC Class Codes: G01N 2035/00237 Handling microquantities of analyte, e.g. microvalves, capillary networks

Abstract

The invention relates to one or more size-based separation modules adapted to increase a concentration of a first analyte in a sample by at least 10,000 fold, wherein said first analyte has an initial concentration in said sample of less than 1×10⁻³ analytes/μL, and an analyzer for analyzing said first analytes in an enriched medium.

Claims

1. A system comprising:

one or more size-based separation modules adapted to increase a concentration of a first analyte in a sample by at least 10,000 fold, wherein said first analyte has an initial concentration in said sample of less than 1×

10<sup>−

3</sup> analytes/μ

L, and

an analyzer comprising computer executable logic for analyzing said first analytes in an enriched medium.

2. The system of claim 1 wherein said analyzer further comprises a microscope, a microarray, or cell counter.

3. The system of claim 1 wherein said computer executable logic detects a color change in the presence of fetal hemoglobin, globin γ

, globin ε

, GPA, i-antigen, CD36, selectins, CD71 or a combination thereof.

4. The system of claim 1 wherein said computer executable logic detects intensities of probes that selectively bind said first analytes.

5. The system of claim 1 wherein said computer executable logic performs three-dimensional image analysis of said first analytes.

6. The system of claim 1 wherein said analyzer has dual scanning capabilities.

7. The system of claim 1 wherein said computer executable logic images said first analytes.

8. The system of claim 1 wherein said computer executable logic determines trisomy, sex of a fetus, or chromosomal abnormalities in a cell of interest.

9. The system of claim 1 wherein each of said size-based separation modules comprises a two-dimensional array of obstacles that direct said first analyte deterministically in a first direction and a second analyte having a hydrodynamic size smaller than said first analyte in a second direction.

10. The system of claim 9 wherein said first analyte is a nucleated red blood cell.

11. The system of claim 9 wherein said first analyte is a fetal nucleated red blood cell.

12. The system of claim 1 wherein said system comprises two or more size-based separation modules fluidly coupled in parallel with one another.

13. The system of claim 1 adapted for high-throughput analysis of at least 10 mL of said fluid sample per hour.

14. The system of claim 1 further comprising one or more capture modules fluidly coupled to said size-based separation modules, wherein said capture modules selectively capture said first analyte or a second analyte from said sample.

15. The system of claim 14 wherein each of said capture modules comprises a two-dimensional array of obstacles.

16. The system of claim 14 wherein each of said capture modules comprises a two-dimensional array of obstacles coupled to one or more antibodies.

17. The system of claim 16 wherein said antibodies selectively bind a red blood cell, a white blood cell, a fetal blood cell, a fetal nucleated blood cell, a cancer cell, an epithelial cell, or stem cell, a progenitor cell, a foam cell, or a platelet.

18. The system of claim 16 wherein said antibodies are selected from the group consisting of:

anti-CD71, anti-CD36, anti-CD451, anti-GPA, anti-selectin, anti-carbohydrates, anti-antigen-i, and anti-EpCaM.

19. The system of claim 1 wherein said fluid sample is a blood sample and said first analyte is selected from the group consisting of:

an epithelial cell, an endothelial cell, a progenitor cell, a stem cell, a foam cell, or a cancer cell.

20. The system of claim 1 wherein said fluid sample is a blood sample and is less than 5 mL.

21. The system of claim 1 wherein said fluid sample is a blood sample derived from a female who is in less than 12 weeks of gestation.

22. The system of claim 1 wherein gap between obstacles in said separation regions is less than 1000 microns.

23. The system of claim 1 further comprising a reservoir containing magnetic particles fluidly coupled to said separation region or said capture regions.

24. The system of claim 1 wherein said system further comprises magnetic beads adapted to preferentially bind to said first analyte or said second analyte.

25. The system of claim 24 wherein said magnetic beads are coupled to an antibody that specifically binds said first analyte.

26. The system of claim 1 further comprising a reservoir fluidly coupled to said size-based separation module, wherein said reservoir contains magnetic particles adapted to preferentially bind said first analyte.

27. The system of claim 2 wherein said analyzer comprises a microarray adapted to detect one or more SNPs in said first analyte.

28. The system of claim 2 wherein said analyzer comprises a microarray adapted to detect levels of mRNA in said first analyte.

29. The system of claim 9 wherein said obstacles are separated by gaps which direct the flow of sample unequally into subsequent gaps.