Microfluidic devices and methods for separating and detecting constituents in a fluid sample

Patent metadata

Field	Value
Publication number	US-9523684-B2
Application number	US-201514798103-A
Country	US
Kind code	B2
Filing date	Jul 13, 2015
Priority date	Jun 24, 2011
Publication date	Dec 20, 2016
Grant date	Dec 20, 2016

Abstract

Microfluidic devices and methods for using the same are provided. Aspects of the present disclosure include microfluidic devices that include a separation medium having functional groups which covalently bond to one or more analytes of interest, e.g., proteins, in a sample upon application of an applied stimulus, e.g., light. Also provided are methods of using the devices as well as systems and kits that include the devices. The devices, systems and methods find use in a variety of different applications, including diagnostic and validation assays.

First claim

That which is claimed is:

1. A microfluidic device for separating one or more constituents of interest in a fluid sample, comprising:
(a) a flow path for permitting said fluid sample having said one or more constituents of interest to flow therethrough; and
(b) a separation medium in the flow path comprising (i) a polymeric gel that permits said fluid sample having said one or more constituents of interest to traverse said flow path after application of a stimulus, and (ii) functional groups attached to said polymeric gel of said separation medium, wherein upon activation by said stimulus, said functional groups covalently bond to said one or more constituents of interest in said separation medium at a capture efficiency of 1% or more for said one or more constituents of interest.

2. The device of claim 1 , wherein said stimulus is electromagnetic radiation.

3. The device of claim 1 , wherein said polymeric gel comprises a pH gradient.

4. The device of claim 1 , wherein said polymeric gel comprises a pore size gradient.

5. The device of claim 1 , further comprising at least one linker group having a first end bound to said functional groups and a second end bound to said separation medium.

6. The device of claim 5 , wherein said at least one linker group comprises a C1-C6 alkyl group.

7. The device of claim 1 , wherein said functional groups comprise N-(3-[(4-benzoylphenyl) formamido]propyl) methacrylamide.

8. The device of claim 1 , wherein said functional groups are co-polymerized with said separation medium.

9. The device of claim 1 , wherein said polymeric gel is a cross-linked polymeric gel.

10. The device of claim 1 , wherein said capture efficiency is about 5% or more.

11. The device of claim 1 , wherein said separation medium comprises pores with pore sizes that depend on a total polymer content of said polymeric gel.

12. The device of claim 11 , wherein said total polymer content is from about 1% to 20%.

13. A method for separating one or more constituents of interest in a fluid sample, comprising:
(a) providing a microfluidic device comprising:
1. a flow path for permitting said fluid sample having said one or more constituents of interest to flow therethrough; and
2. a separation medium in the flow path comprising (i) a polymeric gel that permits said fluid sample having said one or more constituents of interest to traverse said flow path after application of a stimulus, and (ii) functional groups attached to said polymeric gel of said separation medium, wherein upon activation by said stimulus, said functional groups covalently bond to said one or more constituents of interest in said separation medium;

(b) subjecting said fluid sample to flow through said flow path; and
(c) applying said stimulus to said polymeric gel to activate said functional groups to capture said one or more constituents of interest in said separation medium at a capture efficiency of 1% or more.

14. The method of claim 13 , wherein said one or more constituents of interest are from about 2 to about 100 distinct analytes.

15. The method of claim 13 , wherein said fluid sample comprises 10 or more distinct molecular entities.

16. The method of claim 13 , wherein said fluid sample has a volume of 20 microliters or less.

17. The method of claim 13 , further comprising characterizing said one or more constituents of interest.

18. The method of claim 13 , wherein said subjecting comprises use of a flow field to direct said one or more constituents of interest through said flow path.

19. The method of claim 18 , wherein said flow field directs said one or more constituents of interest substantially in the same direction through said flow path.

20. The method of claim 18 , wherein said flow field is an electric field or a pressure differential.

21. The method of claim 13 , wherein said subjecting comprises use of electroosmosis.

22. The method of claim 13 , wherein said one or more constituents of interest are separated in said separation medium by size or molecular mass.

23. The method of claim 13 , wherein said one or more constituents of interest are labeled.

24. A system for separating one or more constituents of interest in a fluid sample, comprising:
(a) a microfluidic device comprising:
1. a flow path for permitting said fluid sample having said one or more constituents of interest to flow therethrough; and
2. a separation medium in the flow path comprising (i) a polymeric gel that permits said fluid sample having said one or more constituents of interest to traverse said flow path after application of a stimulus, and (ii) functional groups attached to said polymeric gel of said separation medium, wherein upon activation by said stimulus, said functional groups covalently bond to said one or more constituents of interest in said separation medium at a capture efficiency of 1% or more for said one or more constituents of interest; and

(b) a source of said stimulus, which source directs said stimulus to said polymeric gel.

25. The system of claim 24 , wherein said source provides electromagnetic radiation.

26. The system of claim 24 , wherein said separation medium further comprises at least one linker group having a first end bound to said functional groups and a second end bound to said separation medium.

27. The system of claim 24 , wherein said capture efficiency is about 5% or more.

Assignees

• Univ California

Inventors

• Herr Amy E
• Hughes Alex James

Classifications

• G01N33/559 Primary

  through a gel, e.g. Ouchterlony technique · CPC title

• G01N33/561

  Immunoelectrophoresis · CPC title

• G01N33/54366

  Apparatus specially adapted for solid-phase testing · CPC title

• G01N27/44756

  Apparatus specially adapted therefor · CPC title

• B01L2400/0496

  Travelling waves, e.g. in combination with electrical or acoustic forces · CPC title