Microfluidic apparatus and method for purification and processing

• US 6,613,525 B2
• Filed: 01/17/2002
• Issued: 09/02/2003
• Est. Priority Date: 07/30/1996
• Status: Expired due to Term

• Current Assignee: Monogram Biosciences, Inc. (Laboratory Corporation Of America Holdings)
• Original Assignee: Aclara Biosciences, Inc (Laboratory Corporation Of America Holdings)
• Inventors: Nelson, Robert J., Williams, Stephen J., Hooper, Herbert H., Hauser, Alan K., Sassi, Alexander P., Singh, Sharat
• Primary Examiner(s): Whisenant, Ethan
• Assistant Examiner(s): Tung, Joyce
• Application Number:
  US10/051,847
• Publication Number:
  US 20020119482A1
• Time in Patent Office: 593 Days
• Field of Search: 435/6, 435/91.1, 435/283.1, 435/285.2, 435/288.5, 435/91.2, 204/601, 204/450, 204/600, 204/451, 204/453
• US Class Current: 435/6.19
• CPC Class Codes: B01F 33/30 Micromixers
• CPC Class Codes: B01L 2200/0631 Purification arrangements, e.g. solid phase extraction [SPE]
• CPC Class Codes: B01L 2200/0668 Trapping microscopic beads
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0861 Configuration of multiple channels and/or chambers in a single devices
• 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 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2300/0877 Flow chambers
• CPC Class Codes: B01L 2400/0409 centrifugal forces
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 2400/0421 electrophoretic flow
• CPC Class Codes: B01L 3/50273 characterised by the means or forces applied to move the fluids micropumps F04B19/006, of the membrane type F04B43/043
• 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 7/52 with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples amplification or hybridisation processes per se C12Q1/68; controlling sequential reactions for synthesis B01J19/0046
• CPC Class Codes: G01N 27/44743 Introducing samples
• CPC Class Codes: G01N 27/44791 Microapparatus sample containers with integrated microfluidic structures B01L3/5027
• CPC Class Codes: G01N 33/5002 Partitioning blood components
• CPC Class Codes: G01N 33/569 for microorganisms, e.g. protozoa, bacteria, viruses

Abstract

Integrated microfluidic devices comprising at least an enrichment channel (10) and a main electrophoretic flowpath (12) are provided. In the subject integrated devices, the enrichment channel and the main electrophoretic flowpath are positioned so that waste fluid flows away from said main electrophoretic flowpath through a discharge outlet (6). The subject devices find use in a variety of electrophoretic applications, including clinical assays, high throughput screening for genomics and pharmaceutical applications, point-or-care in vitro diagnostics, molecular genetic analysis and nucleic acid diagnostics, cell separations, and bioresearch generally.

Claims

1. An integrated microfluidic device having at least one microchannel formed in a generally planar substrate, said device comprising:

at least one microchannel comprising an enrichment channel portion having at least an inlet and an outlet; and

an enrichment medium present in said enrichment channel portion and containing specific binding moieties which bind to a target in a sample such that at least a fraction of the target is retained by said enrichment medium;

said enrichment channel portion is configured to (i) receive via said inlet the target containing sample, and (ii) permit movement of at least a portion of the bound target via said outlet.

6. The device of claim 1, wherein the enrichment medium is present within the enrichment channel portion prior to moving the sample into the enrichment channel portion.

7. The device of claim 1, wherein said target containing sample and said enrichment medium are moved into the enrichment channel portion concurrently.

8. The device of claim 1, wherein the enrichment medium comprises beads coated with specific binding moieties.

9. The device of claim 8, wherein the specific binding moieties are antibodies.

10. The device of claim 8, further including a confining element for immobilizing the beads within the enrichment channel portion.

11. The device of claim 10, where said confining element is selected from the group consisting of at least one magnet, a membrane, a mesh screen and an agarose gel plug.

12. The device of claim 1, further comprising a component capable of eluting bound target from said enrichment medium.

13. The device of claim 1, wherein when said target is a cell, the device further comprises a component capable of lysing said cell.

2. An integrated microfluidic device comprising microchannels formed in a generally planar substrate, said device comprising:

an enrichment microchannel comprising an enrichment channel portion containing an enrichment medium, said enrichment channel portion having a first inlet and a first fluid outlet;

a main electrophoretic microchannel enclosing a main electrophoretic flowpath, said main electrophoretic microchannel being connected in fluid-conducting relation to said first fluid outlet, and said main electrophoretic microchannel comprising a detection region;

means for moving a sample into said enrichment channel portion by way of said first inlet and out from said enrichment channel portion by way of said first fluid outlet, whereby at least a fraction of a component of interest in the sample is retained by said enrichment medium,

a discharge channel connected in fluid communication with said first fluid outlet at a point upstream from said main electrophoretic flowpath, for discharging a portion of said sample other than said fraction away from said main electrophoretic flowpath; and

means for moving at least a portion of the fraction out from said enrichment channel portion by way of said first fluid outlet and into said main electrophoretic microchannel.

3. The device according to claim 2, further comprising a secondary microchannel connected in fluid conducting relation between said first fluid outlet and said main electrophoretic microchannel.

4. The device according to claim 3 wherein said secondary microchannel and said main electrophoretic microchannel intersect to form an injection cross.

5. An integrated microfluidic device comprising microchannels formed in a generally planar substrate, said device comprising:

an enrichment microchannel comprising an enrichment channel portion containing an enrichment medium, said enrichment channel portion having a first inlet and a first fluid outlet;

a main electrophoretic microchannel enclosing a main electrophoretic flowpath, said main electrophoretic microchannel being connected in fluid-conducting relation to said first fluid outlet, and said main electrophoretic microchannel comprising a detection region;

means for moving a sample into said enrichment channel portion by way of said first inlet and out from said enrichment channel portion by way of said first fluid outlet, whereby at least a fraction of a component of interest in the sample is retained by said enrichment medium;

a discharge channel connected in fluid communication with said first fluid outlet at a point upstream from said main electrophoretic flowpath, for discharging a portion of said sample other than said fraction away from said main electrophoretic flowpath;

means for moving at least a portion of the fraction out from said enrichment channel portion by way of said first fluid outlet and into said main electrophoretic microchannel; and

a plurality of input microchannels connected in fluid relation to said first inlet, said plurality of input microchannels comprising at least a sample input microchannel and an elution input microchannel.

14. A method of enriching a target in a sample comprising:

providing a microfluidic device having at least one microchannel formed in a generally planar substrate, wherein at least one of the microchannels comprises an enrichment channel portion having at least an inlet and an outlet;

loading the sample into said enrichment channel portion through said inlet;

loading at least one reagent including an enrichment medium containing specific binding moieties which bind to the target into said enrichment channel portion;

capturing said targets with said specific binding moieties to form a complex between said target and said specific binding moieties; and

washing said enrichment channel portion to remove non-bound sample.

15. The method of claim 14, whereby said capturing is performed by incubating said sample and said enrichment medium until a complex is formed between at least a portion of the target and the specific binding moieties.

16. The method of claim 14, further comprising after said capturing step, immobilizing said target-specific binding moiety complex in the enrichment channel portion.

17. The method of claim 16, further comprising after said washing step, releasing said immobolized target-specific binding moiety complex to form a suspension of the unbound enrichment medium and the complex in the enrichment channel portion.

18. The method of claim 14, further comprising after said washing step, releasing said targets from said complex with an elution buffer.

19. The method of claim 18, further comprising transporting said released target to a main electrophoretic channel located downstream of said enrichment channel portion.

20. The method of claim 19, further comprising after said transporting step, processing said target to detect or analyze said target.

21. The method of claim 14, wherein said target is a cell, the method further comprising lysing said target cells following said capture step.

22. The method of claim 14, wherein said enrichment medium comprises beads coated with specific binding moieties.

23. The method of claim 14, wherein said sample comprises heterogeneous cells and said target is a specific cell type of said heterogeneous cells.

24. The method of claim 14, further comprising performing said washing step at least two times.

25. The method of claim 14, further comprising after said washing step, incubating said complex with an elution buffer until said target is disassociated from said specific binding moieties.

26. The method of claim 14, further comprising after said washing step, processing said target to detect or analyze said target.