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

• US 9,956,562 B2
• Filed: 11/02/2015
• Issued: 05/01/2018
• Est. Priority Date: 04/05/2005
• Status: Active Grant

• Current Assignee: The General Hospital Corporation (Mass General Brigham, Inc.), ZEON Corporation
• Original Assignee: Gpb Scientific, LLC, The General Hospital Corporation (Mass General Brigham, Inc.)
• Inventors: Huang, Lotien Richard, Barber, Thomas A., Carvalho, Bruce L., Kapur, Ravi, Vernucci, Paul, Toner, Mehmet, Wang, Zihua
• Primary Examiner(s): Brown, Melanie Yu
• Application Number:
  US14/930,313
• Publication Number:
  US 20160144378A1
• Time in Patent Office: 911 Days
• 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 devices and methods for the deterministic separation of particles. Exemplary methods include the enrichment of a sample in a desired particle or the alteration of a desired particle in the device. The devices and methods are advantageously employed to enrich for rare cells, e.g., fetal cells, present in a sample, e.g., maternal blood and rare cell components, e.g., fetal cell nuclei. The invention further provides a method for preferentially lysing cells of interest in a sample, e.g., to extract clinical information from a cellular component, e.g., a nucleus, of the cells of interest. In general, the method employs differential lysis between the cells of interest and other cells (e.g., other nucleated cells) in the sample.

Claims

1. A method of producing a sample enriched in particles of a specific type from a liquid sample, the method comprising

obtaining a liquid sample comprising particles of the specific type;

increasing a hydrodynamic size of particles of the specific type in the liquid sample by adding to the liquid sample a reagent that swells the particles of the specific type; and

flowing the liquid sample through a channel under laminar flow conditions, wherein the channel comprises an inlet, an array of obstacles that form a network of gaps, and first and second outlets, wherein any particles in the liquid sample passing into the inlet and through the network of gaps are divided unequally into a major flux and a minor flux so that particles having a hydrodynamic size above a critical size are displaced laterally in the network of gaps to the first outlet and particles having a hydrodynamic size below the critical size are not displaced laterally in the network of gaps and flow to the second outlet, wherein the specific type of particles having an altered hydrodynamic size are directed selectively to either the first outlet or the second outlet, thereby producing a sample enriched in particles of the specific type.

2. The method of claim 1, further comprising separating particles of the specific type from the enriched liquid sample.

3. The method of claim 1, wherein the reagent comprises a hypotonic solution.

4. A method of producing a sample enriched in particles of a specific type from a liquid sample, the method comprising

obtaining a liquid sample comprising particles of the specific type;

decreasing a hydrodynamic size of the particles of the specific type in the liquid sample; and

flowing the liquid sample through a channel under laminar flow conditions, wherein the channel comprises an inlet, an array of obstacles that form a network of gaps, and first and second outlets, wherein any particles in the liquid sample passing into the inlet and through the network of gaps are divided unequally into a major flux and a minor flux so that particles having a hydrodynamic size above a critical size are displaced laterally in the network of gaps to the first outlet and particles having a hydrodynamic size below the critical size are not displaced laterally in the network of gaps and flow to the second outlet, wherein the specific type of particles having an altered hydrodynamic size are directed selectively to either the first outlet or the second outlet, thereby producing a sample enriched in particles of the specific type.

5. The method of claim 4, wherein decreasing the hydrodynamic size comprises adding to the liquid sample a hypertonic solution that shrinks the particles of the specific type.

6. The method of claim 4, further comprising separating particles of the specific type from the enriched liquid sample.

7. A method of producing a sample enriched in particles of a specific type from a liquid sample, the method comprising

obtaining a liquid sample comprising particles of the specific type;

altering a hydrodynamic size of particles of the specific type in the liquid sample; and

flowing the liquid sample through a channel under laminar flow conditions, wherein the channel comprises an inlet, an array of obstacles that form a network of gaps, and first and second outlets, wherein the channel is bounded by two side walls and further comprises a bypass channel defined by a region within the channel devoid of obstacles between a side wall and the array of obstacles, wherein any particles in the liquid sample passing into the inlet and through the network of gaps are divided unequally into a major flux and a minor flux so that particles having a hydrodynamic size above a critical size are displaced laterally in the network of gaps toward the bypass channel and to the first outlet and particles having a hydrodynamic size below the critical size are not displaced laterally in the network of gaps and flow to the second outlet, and wherein the specific type of particles having an altered hydrodynamic size are directed selectively to either the first outlet or the second outlet, thereby producing a sample enriched in particles of the specific type.

8. The method of claim 7, wherein altering the hydrodynamic size comprises increasing the hydrodynamic size of the particles of the specific type.

9. The method of claim 8, wherein increasing the hydrodynamic size comprises adding to the liquid sample microparticles that bind selectively to the particles of the specific type.

10. The method of claim 9, wherein the microparticles comprise an antibody or one of an avidin-biotin binding ligand pair to bind selectively to the particles of the specific type.

11. The method of claim 10, wherein the microparticles are magnetic or polymer beads.

12. The method of claim 8, wherein increasing the hydrodynamic size comprises adding to the liquid sample a reagent that swells the particles of the specific type.

13. The method of claim 12, wherein the reagent comprises a hypotonic solution.

14. The method of claim 7, wherein the particles of the specific type are rare cells.

15. The method of claim 14, wherein the rare cell is a blood cell, a fetal cell, a nucleated red blood cell, a stem cell, a cancer cell, an immune system cell, an epithelial cell, a connective tissue cell, a bacterial cell, a fungal cell, or a parasite.

16. The method of claim 14, wherein the rare cells are blood cells, wherein the channel comprises a second array of obstacles that form a network of gaps and a third outlet, and wherein the method produces three separate sample fractions, wherein a first fraction is enriched in platelets, a second fraction is enriched in red blood cells, and a third fraction is enriched in white blood cells, all from the same liquid sample.

17. The method of claim 14, wherein the rare cells are white blood cells and the sample is enriched in white blood cells.

18. The method of claim 14, wherein the rare cells are platelets and the sample is enriched in platelets.

19. The method of claim 14, wherein the rare cells are red blood cells and the sample is enriched in red blood cells.

20. The method of claim 7, wherein the liquid sample comprises a bodily fluid or an environmental liquid.

21. The method of claim 20, wherein the bodily fluid comprises sweat, tears, ear flow, sputum, lymph, bone marrow suspension, cerebrospinal fluid, urine, saliva, semen, vaginal flow, cerebrospinal fluid, brain fluid, ascites, milk, secretions of the respiratory, intestinal or genitourinary tract, liquefied tissue sample, or amniotic fluid.

22. The method of claim 20, wherein the rare cells are fetal cells and the liquid sample comprises maternal blood.

23. The method of claim 20, wherein the rare cells are circulating cancer cells and the liquid sample comprises blood.

24. The method of claim 7, wherein the particles of the specific type comprise cellular components, viruses, nucleic acids, lipids, proteins, protein complexes, or carbohydrates.

25. The method of claim 7, wherein the array of obstacles comprises at least a first row and a second row, wherein the second row is displaced laterally relative to the first row so that particles in the liquid sample passing through gaps between obstacles in the first row are divided unequally to pass through two gaps between obstacles in the second row.

26. The method of claim 7, further comprising separating particles of the specific type from the enriched liquid sample.