Microfluidic chromatography

• US 20050000900A1
• Filed: 06/21/2004
• Published: 01/06/2005
• Est. Priority Date: 04/06/2001
• Status: Active Grant

• Current Assignee: Fluidigm Corporation
• Original Assignee: Fluidigm Corporation
• Inventors: Unger, Marc A., Chou, Hou-Pu, Huang, Jiang
• Granted Patent: US 7,217,367 B2
• Time in Patent Office: Days
• US Class Current: 210/656
• CPC Class Codes: B01D 15/08 Selective adsorption, e.g. chromatography
• CPC Class Codes: G01N 2030/387 Turbulent flow of mobile phase
• CPC Class Codes: G01N 2030/521 form
• CPC Class Codes: G01N 30/6065 with varying cross section
• CPC Class Codes: G01N 30/6095 Micromachined or nanomachined, e.g. micro- or nanosize
• CPC Class Codes: Y10T 137/86863 Rotary valve unit
• CPC Class Codes: Y10T 436/2575 Volumetric liquid transfer

Abstract

The present invention is directed to a microfluidic chromatography apparatus comprising a microfabricated fluid delivery system and a chromatography column which is in fluid communication with the fluid delivery system, and a method for producing and using the same. Preferably, the chromatography column comprises an OTLC, PCLC, or combinations thereof.

Claims

1-14. -14. (Canceled)

15. A method for producing a microfluidic chromatography apparatus comprising:

(a) producing a microfabricated fluid delivery system from a material comprising an elastomeric polymer, wherein the fluid deliver system comprises;

(i) a microfluidic flow channel comprising a flow channel inlet for introducing the fluid into said flow channel and a flow channel outlet,

(ii) a flow control channel,

(iii) a flow control valve comprised of a flow control elastomeric segment that is disposed in between said flow channel and said flow control channel to regulate fluid flow through said flow channel, wherein said flow control valve is deflectable into or retractable from said flow channel upon which said flow control valve operates in response to an actuation force applied to said flow control channel, said flow control elastomeric segment when positioned in said flow channel restricting fluid flow therethrough, and

(iv) a flow control channel actuation system operatively interconnected to said flow control channel for applying an actuation force to said flow control channel; and

(b) connecting the fluid delivery system to a chromatography column having a column inlet and a column outlet such that the column inlet is in fluid communication with the flow channel outlet, wherein the chromatography column comprises a stationary phase which is capable of separating at least a portion of the analyte in the fluid.

16. The method of claim 15, wherein the flow channel is located on an interface between a solid substrate and the elastomeric polymer such that an inner surface of the flow channel comprises an elastomeric polymer portion and a solid substrate portion.

17. The method of claim 16, wherein the stationary phase is attached to the solid substrate portion of the flow channel inner surface.

18. The method of claim 17, wherein the elastomeric polymer portion of the flow channel inner surface comprises a surface coating that reduces a non-specific binding of the analyte.

19. The method of claim 15 further comprising:

(a) microfabricating the chromatography column which comprises a chromatography channel having an inner surface which comprises a functional group; and

(b) attaching a stationary phase compound to at least a portion of the inner surface by reacting the stationary phase compound with the functional group under conditions sufficient to form a covalent bond between the functional group and the stationary phase compound.

20. The method of claim 19, wherein the functional group is silane.

21. The method of claim 20, wherein the stationary phase compound is 1-octadecene.

22. The method of claim 19, wherein the microfabricated chromatography column further comprises a microfabricated rotary channel comprising:

a rotary channel inlet;

a rotary channel outlet;

a rotary control channel;

a rotary inlet control valve comprised of an elastomeric segment of said rotary inlet control channel that is disposed in between said rotary channel inlet and said rotary control channel to regulate fluid flow into said rotary channel, wherein said rotary inlet control valve is deflectable into or retractable from said rotary channel inlet upon which said rotary inlet control valve operates in response to an actuation force applied to said rotary control channel, said elastomeric segment of said rotary inlet control channel when positioned in said rotary channel inlet restricting fluid flow therethrough;

a rotary outlet control valve comprised of an elastomeric segment of said rotary outlet control channel that is disposed in between said rotary channel outlet and said rotary control channel to regulate fluid flow out of said rotary channel, wherein said rotary outlet control valve is deflectable into or retractable from said rotary channel outlet upon which said rotary outlet control valve operates in response to an actuation force applied to said rotary control channel, said elastomeric segment of said rotary control channel outlet when positioned in said rotary channel outlet restricting fluid flow therethrough;

a rotary pump valve comprised of an elastomeric segment of said rotary pump that is disposed in between said rotary channel and said rotary pump control channel to regulate fluid flow through said rotary channel, wherein said rotary pump valve is deflectable into or retractable from said rotary channel upon which said rotary pump valve operates in response to an actuation force applied to said rotary pump control channel, said elastomeric segment of said rotary pump when positioned in said rotary channel restricting fluid flow therethrough; and

a rotary control channel actuation system operatively interconnected to said rotary control channel for applying an actuation force to said rotary control channel.

23. A method for separating an analyte from a sample fluid comprising:

(A) introducing the sample fluid into a microfluidic chromatography apparatus comprising;

(a) a microfabricated fluid delivery system which is produced from a material comprising an elastomeric polymer, wherein said fluid deliver system comprises;

(i) a microfluidic flow channel comprising a flow channel inlet for introducing the fluid into said flow channel and a flow channel outlet,

(ii) a flow control channel,

(iii) a flow control valve comprised of a flow control elastomeric segment that is disposed in between said flow channel and said flow control channel to regulate fluid flow through said flow channel, wherein said flow control valve is deflectable into or retractable from said flow channel upon which said flow control valve operates in response to an actuation force applied to said flow control channel, said flow control elastomeric segment when positioned in said flow channel restricting fluid flow therethrough, and

(iv) a flow control channel actuation system operatively interconnected to said flow control channel for applying an actuation force to said flow control channel; and

(b) a chromatography column comprising

(i) a stationary phase which is capable of separating at least a portion of the analyte from the sample fluid,

(ii) a column inlet which is in fluid communication with the flow channel outlet, and

(iii) a column outlet through which a separated fluid exits the chromatography column; and

(B) eluting the sample fluid through the chromatography column with an eluent to separate at least a portion of the analyte.

24. The method of claim 23, wherein fluid flow through the chromatography column is achieved by a peristaltic pump action created by actuating one or more of the flow control valves.

25. The method of claim 24, wherein the chromatography column comprises a microfabricated rotary channel comprising:

a rotary channel inlet;

a rotary channel outlet;

a rotary control channel;

a rotary inlet control valve comprised of an elastomeric segment of said rotary inlet control channel that is disposed in between said rotary channel inlet and said rotary control channel to regulate fluid flow into said rotary channel, wherein said rotary inlet control valve is deflectable into or retractable from said rotary channel inlet upon which said rotary inlet control valve operates in response to an actuation force applied to said rotary control channel, said elastomeric segment of said rotary inlet control channel when positioned in said rotary channel inlet restricting fluid flow therethrough;

a rotary outlet control valve comprised of an elastomeric segment of said rotary outlet control channel that is disposed in between said rotary channel outlet and said rotary control channel to regulate fluid flow out of said rotary channel, wherein said rotary outlet control valve is deflectable into or retractable from said rotary channel outlet upon which said rotary outlet control valve operates in response to an actuation force applied to said rotary control channel, said elastomeric segment of said rotary control channel outlet when positioned in said rotary channel outlet restricting fluid flow therethrough;

a rotary pump valve comprised of an elastomeric segment of said rotary pump that is disposed in between said rotary channel and said rotary pump control channel to regulate fluid flow through said rotary channel, wherein said rotary pump valve is deflectable into or retractable from said rotary channel upon which said rotary pump valve operates in response to an actuation force applied to said rotary pump control channel, said elastomeric segment of said rotary pump when positioned in said rotary channel restricting fluid flow therethrough; and

a rotary control channel actuation system operatively interconnected to said rotary control channel for applying an actuation force to said rotary control channel.

26. The method of claim 25 further comprising:

introducing at least a portion of the sample fluid into the rotary channel;

closing the rotary inlet and the rotary outlet control valves by actuating the rotary inlet and the rotary outlet control valves;

transporting the sample fluid through the rotary channel by actuating one or more of the rotary pump valves until at least a portion of the analyte is adsorbed onto the stationary phase;

opening the rotary inlet and rotary outlet control channels;

introducing a first eluent through the rotary inlet channel and removing the resulting mixture through the rotary outlet channel, whereby substantially all of the sample fluid is removed from the rotary channel and at least about 95% of the adsorbed analyte remains adsorbed onto the stationary phase; and

introducing a second eluent, which is capable of removing the analyte from the stationary phase, through the rotary inlet channel and removing the resulting mixture through the rotary outlet channel, whereby substantially all of the adsorbed analyte is removed from the rotary channel.

27. The method of claim 26, wherein said analyte is a protein having a molecular weight of at least about 1000 g/mol.

28. The method of claim 27, wherein the stationary phase is C-18 alkyl and the sample fluid is an aqueous solution.

29. The method of claim 28, wherein the first eluent is selected from the group consisting of water and an aqueous buffer solution.

30. The method of claim 29, wherein the second eluent comprises an organic solvent selected from the group consisting of an alcohol, acetonitrile, dimethylformamide, and mixtures thereof.

31. The method of claim 30, wherein the second eluent is a mixture of the organic solvent and water or an aqueous buffer solution.