High throughput screening assay systems in microscale fluidic devices

• US 6,558,944 B1
• Filed: 07/01/1999
• Issued: 05/06/2003
• Est. Priority Date: 06/28/1996
• Status: Expired due to Term

• Current Assignee: Caliper Holdings Incorporated
• Original Assignee: Caliper Holdings Incorporated
• Inventors: Bousse, Luc J., Parce, John Wallace, Kopf-Sill, Anne R.
• Primary Examiner(s): Chin, Christopher L.
• Application Number:
  US09/346,660
• Time in Patent Office: 1,405 Days
• Field of Search: 435/6, 435/7.2, 435/7.21, 435/259, 435/291, 435/287.1, 435/287.2, 435/287.9, 435/288.4, 435/288.5, 435/288.7, 435/810, 435/970, 436/164, 436/518, 436/524, 436/527, 436/531, 436/533, 436/534, 436/805, 436/806, 436/807, 436/809, 436/514, 422/55-58
• US Class Current: 435/287.2
• CPC Class Codes: B01J 19/0093 Microreactors, e.g. miniaturised or microfabricated reactors laboratory containers with capillary fluid transport in microfabricated channels or chambers B01L3/5027
• CPC Class Codes: B01L 2200/0605 Metering of fluids
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2200/0668 Trapping microscopic beads
• CPC Class Codes: B01L 2200/0673 Handling of plugs of fluid surrounded by immiscible fluid
• CPC Class Codes: B01L 2200/0689 Sealing
• CPC Class Codes: B01L 2200/12 Specific details about manufacturing devices
• 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 2300/0877 Flow chambers
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 3/5025 for parallel transport of multiple samples
• 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 3/502784 specially adapted for droplet or plug flow, e.g. digital microfluidics automatic analysis using a stream of discrete samples in a tube system G01N35/08
• CPC Class Codes: B01L 9/527 for microfluidic devices, e.g. used for lab-on-a-chip
• CPC Class Codes: G01N 27/44791 Microapparatus sample containers with integrated microfluidic structures B01L3/5027
• CPC Class Codes: G01N 33/5008 : for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
• CPC Class Codes: G01N 33/5011 : for testing antineoplastic activity
• CPC Class Codes: G01N 33/502 : for testing non-proliferative effects
• CPC Class Codes: G01N 33/5044 : involving specific cell types
• CPC Class Codes: G01N 33/5064 : Endothelial cells
• CPC Class Codes: G01N 33/5091 : for testing the pathological state of an organism
• CPC Class Codes: G01N 33/5094 : for blood cell populations red blood cells G01N33/80
• CPC Class Codes: G01N 33/5304 : Reaction vessels, e.g. agglutination plates for solid-phase systems G01N33/543
• CPC Class Codes: G01N 33/57415 : of breast
• CPC Class Codes: G01N 35/08 : using a stream of discrete samples flowing along a tube system, e.g. flow injection analysis
• CPC Class Codes: Y10S 366/02 : Micromixers: segmented laminar flow with boundary mixing orthogonal to the direction of fluid propagation with or without geometry influences from the pathway
• CPC Class Codes: Y10S 435/81 : Packaged device or kit
• CPC Class Codes: Y10S 435/97 : Test strip or test slide
• CPC Class Codes: Y10S 436/805 : Optical property
• CPC Class Codes: Y10S 436/806 : Electrical property or magnetic property
• CPC Class Codes: Y10S 436/807 : Apparatus included in process claim, e.g. physical support structures
• CPC Class Codes: Y10S 436/809 : Multifield plates or multicontainer arrays

Abstract

The present invention provides novel microfluidic devices and methods that are useful for performing high-throughput screening assays. In particular, the devices and methods of the invention are useful in screening large numbers of different compounds for their effects on a variety of chemical, and preferably, biochemical systems.

Claims

1. A method of detecting an interaction between two components of a biochemical system, comprising:

moving a first labeled component of the biochemical system through a first channel region;

moving a second component of the biochemical system through the first channel region to mix with the first component within the first channel region, whereupon the first and second components interact in a continuously flowing mixture to produce a labeled product, which labeled product has a mobility through the first channel region which is substantially equal to a mobility of the first labeled component through the first channel region;

moving the labeled product through a second channel region connected to the first channel region, the labeled product having a mobility through the second channel region which is different from a mobility of the first labeled component through the second channel region; and

detecting an amount of interaction between the first and second components by detecting an amount of labeled product in thy second channel region.

2. The method of claim 1, wherein the product has a different electrokinetic mobility from the first and second components, and the method further comprises applying an electric field along at least the second channel region.

3. The method of claim 1, wherein the first component comprises a first member of a receptor-ligand pair, and the second component comprises a second member of the receptor-ligand pair, and the product comprises a receptor ligand complex.

4. The method of claim 1, wherein the first component comprises a first member of an antibody-antigen pair, the second component comprises a second member of the antibody-antigen pair, and the product comprises an antibody antigen complex.

5. The method of claim 1, wherein the first component comprises a first nucleic acid and the second component comprises a second nucleic acid that is complementary to the first nucleic acid.

6. The method of claim 1, wherein the first component comprises a substrate, the second component comprises an enzyme and the product comprises a product of an interaction of the substrate and the enzyme.

7. The method of claim 1, wherein the product has a different charge from the first component.

8. The method of claim 1, wherein the product has a different size than the first component.

9. The method of claim 1, further comprising contacting the first and second components with a test compound, and detecting an amount of product produced with and without the test compound being present.

10. The method of claim 1, further comprising contacting the first and second components with a test compound, and detecting a decrease in an amount of first labeled component with and without the test compound being present.

11. The method of claim 9, further comprising determining an effect, if any, of the test compound on the interaction between the first and second components, by comparing an amount of product produced without the test compound being present to an amount of product produced with the test compound being present, an increase or decrease in the amount of product produced with the test compound being present being indicative that the test compound is an enhancer or an inhibitor of the interaction of the first and second components.

12. The method of claim 9, further comprising determining an effect, if any, of the test compound on the interaction the first and second component, by comparing a decrease in an amount of the first labeled component without the test compound being present to a decrease in an amount of the first labeled component with the test compound being present, a greater or lesser decrease in the amount of the first labeled component with the test compound being present being indicative that the test compound is an inhibitor or an enhancer of the interaction of the first and second components, respectively.

13. The method of claim 9, wherein the first and second components are continuously flowed through the first and second channel regions, and the test compound is contacted with the first and second components by introducing a discrete plug of the test compound into the first channel region.

14. The method of claim 13, wherein a plurality of different test compounds are separately introduced into the first channel region to contact the first and second components flowing therethrough.

15. The method of claim 13, further comprising detecting a deviation in an amount of product produced with the test compound being present relative to amount of product produced without the test compound being present.

16. The method of claim 13, further comprising detecting a deviation in an amount of decrease in a level of the first labeled component with the test compound being present relative to an amount of decrease of the first labeled component without the test compound being present.

17. The method of claim 1, wherein the second component is flowed into the first channel region from a side channel that intersects the first channel region.

18. The method of claim 1, wherein the test compound is introduced into the first channel region from a side channel that intersects the first channel.

19. The method of claim 18, wherein the side channel is fluidly connected to a pipettor capillary channel that is disposed through a capillary element that is attached to the body of the device.

20. The method of claim 1, wherein the labeled product comprises a fluorescent label.

21. The method of claim 1, wherein the first labeled component comprises a fluorescent label.

22. The method of claim 1, wherein the first component, second component and product are flowed through the first and second channel regions by pumping.

23. The method of claim 1, wherein the labeled product has a mobility in the second channel region that is faster than the mobility of the first labeled component.

24. The method of claim 1, wherein the labeled product has a flow rate that is slower than the mobility of the first labeled component in the second channel region.

25. A method of detecting an interaction between two components of a biochemical system, comprising:

providing a microfluidic device having a body and first and second interconnected channel regions disposed therein;

moving a first component of the biochemical system through the first channel region;

moving a second component of the biochemical system through the first channel region to mix with the first component within the first channel region, whereupon the first and second components interact in a continuously flowing mixture to produce a product, the product having a mobility through the first channel region which is substantially equal to a mobility of the first and second components through the first channel region;

a mobility through the second channel region which is different from a mobility of at least one of the first second components through the second channel region;

moving the product through the second channel region; and

detecting the interaction of the first and second components by virtue of the different mobility of the product relative to the mobility of at least one of the first and second components through the second channel region.

26. A method of detecting interaction between two components of a biochemical system, comprising:

moving a first component of the biochemical system through a channel;

moving a second component of the biochemical system through the channel to mix with the first component within the channel, whereupon the first and second components interact to produce a product, the product having a mobility through the channel that is different from at least one of the first and second components; and

detecting the interaction of the first and second components by virtue of the different mobility of the product relative to the mobility of at least one of the first and second components.

27. A method of detecting an interaction between first and second components of a biochemical system, comprising:

providing a microfluidic device that comprises a body having at least first and second intersecting microscale channels disposed therein, the first and second microscale channels fluidly communicating with a plurality of reservoirs disposed in the body;

placing the first component of the biochemical system into a first reservoir of the microfluidic device;

placing the second component of the biochemical system into a second reservoir of the microfluidic device;

combining the first and second components in a mixture, the first and second components interacting in a continuously flowing mixture to produce a product, the product having a different flow rate through the microfluidic channel than the first and second components;

moving the mixture along the microscale channel; and

detecting an amount of product;

within the microscale channel.