Combinatorial synthesis system

• US 7,097,809 B2
• Filed: 04/03/2002
• Issued: 08/29/2006
• Est. Priority Date: 10/03/2000
• Status: Expired due to Fees

• Current Assignee: California Institute of Technology
• Original Assignee: California Institute of Technology
• Inventors: Van Dam, Michael, Unger, Marc A., Quake, Stephen R.
• Primary Examiner(s): Warden, Jill
• Assistant Examiner(s): SINES, BRIAN J
• Application Number:
  US10/116,761
• Publication Number:
  US 20030008411A1
• Time in Patent Office: 1,609 Days
• Field of Search: 422/50, 422/55, 422/57, 422/58, 422/61, 422/68.1, 422/81, 422/82, 422/82.01, 422/82.02, 422/82.05, 422/100, 422/101, 422/102, 422/103, 422/104, 436/174, 436/178, 436/180, 436/2, 436/8, 436/501, 436/63, 436/66, 436/68
• US Class Current: 422/502
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00355 peristaltic
• CPC Class Codes: B01J 2219/00398 in multiple arrangements
• CPC Class Codes: B01J 2219/00418 using pressure
• CPC Class Codes: B01J 2219/00427 using masks
• CPC Class Codes: B01J 2219/00497 Features relating to the solid phase supports
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/0059 Sequential processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/0061 The surface being organic
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00617 by chemical means
• CPC Class Codes: B01J 2219/00621 by physical means, e.g. trenches, raised areas
• CPC Class Codes: B01J 2219/00626 Covalent
• CPC Class Codes: B01J 2219/00635 by reactive plasma treatment
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00641 the porous medium being continuous, e.g. porous oxide substrates
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00675 : In-situ synthesis on the substrate
• CPC Class Codes: B01J 2219/00722 : Nucleotides
• CPC Class Codes: B01J 2219/00725 : Peptides
• CPC Class Codes: B01J 2219/00729 : Peptide nucleic acids [PNA]
• CPC Class Codes: B82Y 30/00 : Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C40B 40/06 : Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 40/10 : Libraries containing peptides or polypeptides, or derivatives thereof
• CPC Class Codes: C40B 50/14 : Solid phase synthesis, i.e. wherein one or more library building blocks are bound to a solid support during library creation; Particular methods of cleavage from the solid support
• CPC Class Codes: C40B 60/14 : for creating libraries
• CPC Class Codes: G01N 15/06 : Investigating concentration of particle suspensions by weighing G01N5/00; investigating sedimentation of particle suspensions G01N15/04; investigating individual particles G01N15/10
• CPC Class Codes: Y10T 436/10 : Composition for standardization, calibration, simulation, stabilization, preparation or preservation; processes of use in preparation for chemical testing
• CPC Class Codes: Y10T 436/25 : including sample preparation
• CPC Class Codes: Y10T 436/255 : including use of a solid sorbent, semipermeable membrane, or liquid extraction
• CPC Class Codes: Y10T 436/2575 : Volumetric liquid transfer

Abstract

The present invention provides a microfluidic device for synthesizing an array of compounds and methods for using the same. In particular, the microfluidic device of the present invention comprises a solid support base, an elastomeric layer attached to the solid support, and a plurality of flow channels located within the interface between the solid support and the elastomeric layer. In addition, the solid support comprises a functional group for forming a bond with a reactive reagent. In some embodiments, the microfluidic device further comprises a second plurality of flow channels that intersect the first plurality of flow channels. A plurality of control channels are also present in the microfluidic devices of the present invention. The control channels can be actuated to regulate flow of fluids within the flow channel(s).

Claims

1. A microfluidic combinatorial chemical synthesis device, comprising:

(a) a first solid support comprising a plurality of microfluidic flow channels formed therein, wherein a reactive functional group for attaching a reactive reagent thereto is attached to a surface of each of the flow channels;

(b) a second solid support comprising a plurality of control channels formed therein; and

(c) an elastomeric layer sandwiched between the first and second solid support and disposed such that the flow channels and the control channels are separated by different segments of the elastomeric layer, wherein each elastomeric segment is deflectable into or retractable from the fluid flow channel that it overlays in response to an actuation force applied to one of the control channels.

2. The microfluidic combinatorial chemical synthesis device of claim 1, wherein the first solid support is comprised of a material selected from the group consisting of glass, polystyrene, polystyrene-divinylbenzene copolymer, silicone rubber, quartz, latex, polyurethane, a derivatizable transition metal, silicon dioxide, silicon nitride, gallium arsenide, and a derivative thereof.

3. The microfluidic combinatorial chemical synthesis device of claim 2, wherein said derivatizable transition metal is gold.

4. The microfluidic combinatorial chemical synthesis device of claim 1, wherein said first solid support is a rigid material.

5. The microfluidic combinatorial chemical synthesis device of claim 4, wherein said second solid support is made from a rigid material.

6. The microfluidic combinatorial chemical synthesis device of claim 5, wherein said elastomeric layer is removeably disposed in between said first and second solid supports such that said elastomeric layer forms a part of said fluid flow channels and said control channels.

7. The microfluidic combinatorial chemical synthesis device of claim 1, wherein at least a portion of said plurality of control channels are aligned in between said fluid flow channels.

8. The microfluidic combinatorial chemical synthesis device of claim 7, wherein at least a portion of said control channels are aligned on top of said fluid flow channels.

9. The microfluidic combinatorial chemical synthesis device of claim 8, wherein each of said control channels aligned on top of said fluid flow channels comprise regions of wider cross-section and regions of narrower cross-section.

14. The microfluidic combinatorial chemical synthesis device of claim 1, wherein the elastomeric layer is a material that is inert to solvents used in the synthesis of oligonucleotides.

15. The microfluidic combinatorial chemical synthesis device of claim 1, wherein the elastomeric layer is a fluoroelastomer.

10. A method for synthesizing a library of compounds on a microfluidic device, the method comprising:

(a) providing a microfluidic device comprising a plurality of fluid flow channels defined by an elastomeric layer and a solid support such that the flow channels comprise an elastomeric portion and a solid support portion, wherein the solid support comprises a reactive functional group attached to the inner surface of the flow channels for attaching a reactive reagent thereto;

(b) producing a solid-support bound compound by introducing a reactive reagent into the fluid flow channels under conditions sufficient to covalently attach at least a portion of the first reactive reagent to the reactive functional group;

(c) modifying the solid support-bound compound by introducing another reactive reagent into the fluid flow channels under conditions sufficient to react the solid-support bound compound with the reactive reagent; and

(d) optionally repeating step (c), thereby obtaining the library of compounds, and wherein the reactive agent introduced in steps (b)–

(d) is introduced by providing a laminar flow of a sheath fluid in between the reactive agent and the elastomeric layer portion of the fluid flow channels, wherein the sheath fluid is compatible with the elastomeric layer.

11. A method for synthesizing a library of compounds on a microfluidic device, the method comprising:

(a) positioning a solid support and an elastomeric layer having a plurality of recesses formed therein with respect to one another to form a first plurality of flow channels, wherein the solid support comprises a plurality of reactive functional groups for attaching a reactive reagent thereto that are attached to an inner surface of at least some of the first flow channels;

(b) producing a solid-support bound compound by introducing a first reactive reagent into one or more of the fluid flow channels under conditions sufficient to covalently attach at least a portion of the first reactive reagent to the reactive functional group;

(c) repositioning the elastomeric layer with respect to the solid support to form a second plurality of flow channels defined by the solid support and the recesses in the elastomeric layer such that the second flow channels intersect the first flow channels;

(d) modifying the solid support-bound compound by introducing a second reactive reagent into one or more of the second fluid flow channels under conditions sufficient to react, the solid-support bound compound with the second reactive reagent.

12. A method for synthesizing a library of compounds on a microfluidic device, the method comprising:

(a) providing a microfluidic device comprising (i) a first solid support comprising a plurality of microfluidic flow channels formed therein, wherein a reactive functional group for attaching a reactive reagent thereto is attached to a surface of each of the flow channels, (ii) a second solid support comprising a plurality of control channels formed therein; and

(iii) an elastomeric layer sandwiched between the first and second solid support and disposed such that the flow channels and the control channels are separated by different segments of the elastomeric layer, wherein each elastomeric segment is deflectable into or retractable from the fluid flow channel that it overlays in response to an actuation force applied to one of the control channels;

(b) producing a solid-support bound compound by introducing a reactive reagent into the fluid flow channels under conditions sufficient to covalently attach at least a portion of the first reactive reagent to the reactive functional group;

(c) modifying the solid support-bound compound by introducing another reactive reagent into the fluid flow channels under conditions sufficient to react the solid-support bound compound with the reactive reagent; and

(d) optionally repeating step (c), thereby obtaining the library of compounds.

13. The method of claim 12, wherein the fluid flow channels and the control channels are located on the interface of the elastomeric layer and the solid support.