Microfluidic device for cell separation and uses thereof

• US 10,081,014 B2
• Filed: 11/18/2016
• Issued: 09/25/2018
• Est. Priority Date: 09/27/2002
• Status: Expired due to Term

• Current Assignee: The General Hospital Corporation (Mass General Brigham, Inc.)
• Original Assignee: The General Hospital Corporation (Mass General Brigham, Inc.)
• Inventors: Toner, Mehmet, Truskey, George, Kapur, Ravi
• Primary Examiner(s): Ware, Debbie K
• Application Number:
  US15/356,276
• Publication Number:
  US 20170197214A1
• Time in Patent Office: 676 Days
• CPC Class Codes: B01L 1/52 Transportable laboratories; Field kits
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2200/0652 Sorting or classification of particles or molecules
• CPC Class Codes: B01L 2200/0668 Trapping microscopic beads
• CPC Class Codes: B01L 2200/12 Specific details about manufacturing devices
• CPC Class Codes: B01L 2300/0681 Filter
• 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 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2300/0877 Flow chambers
• CPC Class Codes: B01L 2300/0883 Serpentine channels
• CPC Class Codes: B01L 2300/12 Specific details about materials
• 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: B01L 9/54 Supports specially adapted for pipettes and burettes automated pipetting stations G01N35/10
• CPC Class Codes: B33Y 80/00 Products made by additive manufacturing
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C12N 5/0087 : Purging against subsets of blood cells, e.g. purging alloreactive T cells
• CPC Class Codes: G01N 1/40 : Concentrating samples
• CPC Class Codes: G01N 1/405 : by adsorption or absorption
• CPC Class Codes: G01N 2015/1006 : for cytology
• CPC Class Codes: G01N 2015/1028 : Sorting particles
• CPC Class Codes: G01N 2333/70582 : CD71
• CPC Class Codes: G01N 2333/70589 : CD45
• CPC Class Codes: G01N 2333/70596 : Molecules with a "CD"-designation not provided for elsewhere in G01N2333/705
• CPC Class Codes: G01N 33/5091 : for testing the pathological state of an organism
• CPC Class Codes: G01N 33/54366 : Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 33/54386 : Analytical elements
• CPC Class Codes: G01N 33/56966 : Animal cells
• CPC Class Codes: G01N 33/574 : for cancer
• CPC Class Codes: Y10T 29/49982 : Coating
• CPC Class Codes: Y10T 428/24744 : Longitudinal or transverse tubular cavity or cell

Abstract

Methods for separating cells from a sample (e.g., separating fetal red blood cells from maternal blood) are described. The method begins with the introduction of a sample including cells into one or more microfluidic channels. In one embodiment, the device includes at least two processing steps. For example, a mixture of cells is introduced into a microfluidic channel that selectively allows the passage of a desired type of cell, and the population of cells enriched in the desired type is then introduced into a second microfluidic channel that allows the passage of the desired cell to produce a population of cells further enriched in the desired type. The selection of cells is based on a property of the cells in the mixture, for example, size, shape, deformability, surface characteristics (e.g., cell surface receptors or antigens and membrane permeability), or intracellular properties (e.g., expression of a particular enzyme).

Claims

1. A device for separating fetal blood cells from a liquid sample, comprising:

a substrate;

a flow chamber having an inlet and an outlet arranged on the substrate; and

a plurality of obstacles arranged in an array within the flow chamber on the substrate, wherein obstacles in a first region of the flow chamber are coupled to one or more first binding moieties that selectively bind to a first type of blood cell and wherein obstacles in a second region of the flow chamber are coupled to one or more second binding moieties that selectively bind to a second type of blood cell comprising fetal blood cells when a liquid sample comprising fetal blood cells is flowed through the flow chamber.

2. The device of claim 1, wherein the first binding moieties selectively bind to adult blood cells.

3. The device of claim 2, wherein the first binding moieties selectively bind to adult white blood cells.

4. The device of claim 2, wherein the first binding moieties selectively bind to adult red blood cells.

5. The device of claim 2, wherein the first binding moieties selectively bind to maternal blood cells.

6. The device of claim 5, wherein the first binding moieties selectively bind to maternal white blood cells.

7. The device of claim 5, wherein the first binding moieties selectively bind to maternal red blood cells.

8. The device of claim 1, wherein the second binding moieties selectively bind to fetal nucleated red blood cells.

9. The device of claim 1, wherein the second binding moieties selectively bind to fetal white blood cells.

10. The device of claim 1, wherein the first and second binding moieties independently comprise one or more of an antibody, a charged polymer, a molecule that binds to a cell surface receptor, a polypeptide, a viral protein, a bacterial protein, a nucleic acid, or a carbohydrate.

11. The device of claim 10, wherein the first binding moieties are antibodies.

12. The device of claim 11, wherein the antibodies comprise anti-CD45 antibodies that selectively bind to white blood cells.

13. The device of claim 10, wherein the second binding moieties are antibodies.

14. The device of claim 13, wherein the antibodies comprise anti-CD71 antibodies that selectively bind to fetal nucleated red blood cells.

15. The device of claim 1, wherein the first and second regions of obstacles are arranged in series in the flow chamber.

16. The device of claim 1, wherein the first and second regions of obstacles are arranged in parallel in the flow chamber.

17. The device of claim 1, wherein the array of obstacles is an ordered array.

18. The device of claim 17, wherein the obstacles in the first and second regions are arranged in a triangular array.

19. The device of claim 17, wherein the ordered array comprises a plurality of rows of obstacles spaced equally apart within each row, wherein every subsequent row of obstacles is shifted relative to a previous row by a distance of at least one half of a space between adjacent obstacles in the previous row.

20. The device of claim 1, wherein each obstacle, independently, has a cross-sectional shape that is circular, semi-circular, oval, triangular, trapezoidal, rectangular, or square.