Method and apparatus for producing position addressable combinatorial libraries

• US 5,763,263 A
• Filed: 07/24/1996
• Issued: 06/09/1998
• Est. Priority Date: 11/27/1995
• Status: Expired due to Fees

• Current Assignee: Peter J. Dehlinger
• Original Assignee: Peter J. Dehlinger
• Inventors: Dehlinger, Peter J.
• Primary Examiner(s): Hutzell, Paula K.
• Assistant Examiner(s): BAKALYAR, HEATHER
• Application Number:
  US08/686,373
• Time in Patent Office: 685 Days
• Field of Search: 435/287, 435/6, 435/7.1, 530/333, 530/334, 530/25.3, 436/518, 436/527
• US Class Current: 435/6.12
• 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 2219/00286 Reactor vessels with top and bottom openings
• CPC Class Codes: B01J 2219/00315 Microtiter plates
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00333 Closures attached to the reactor vessels
• CPC Class Codes: B01J 2219/00351 Means for dispensing and evacuation of reagents
• CPC Class Codes: B01J 2219/00369 in multiple or parallel arrangements
• CPC Class Codes: B01J 2219/00378 Piezoelectric or ink jet dispensers
• CPC Class Codes: B01J 2219/00511 Walls of reactor vessels
• CPC Class Codes: B01J 2219/0052 in the shape of elongated tubes
• 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 2219/00599 Solution-phase processes
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• 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 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/0072 : Organic compounds
• CPC Class Codes: B01J 2219/00722 : Nucleotides
• CPC Class Codes: B01J 2219/00725 : Peptides
• CPC Class Codes: B01J 2219/00727 : Glycopeptides
• CPC Class Codes: B01J 2219/00731 : Saccharides
• CPC Class Codes: B01L 3/5085 : for multiple samples, e.g. microtitration plates
• CPC Class Codes: C07K 1/047 : Simultaneous synthesis of different peptide species; Peptide libraries
• CPC Class Codes: C12Q 1/6837 : using probe arrays or probe chips C12Q1/6874 takes precedence
• 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 40/12 : Libraries containing saccharides or polysaccharides, or derivatives thereof
• CPC Class Codes: C40B 50/08 : Liquid phase synthesis, i.e. wherein all library building blocks are in liquid phase or in solution during library creation; Particular methods of cleavage from the liquid support
• CPC Class Codes: C40B 60/14 : for creating libraries
• CPC Class Codes: G01N 2001/282 : with mapping; Identification of areas; Spatial correlated pattern
• CPC Class Codes: G01N 2035/00237 : Handling microquantities of analyte, e.g. microvalves, capillary networks
• CPC Class Codes: G01N 35/1065 : Multiple transfer devices

Abstract

Method and apparatus for producing combinatorial position-addressable libraries of different-sequence oligomers or different-substituent small molecule compounds are disclosed. The method employs massive parallel synthesis by stepwise subunit addition or substituent addition in a dense capillary-tube array. The libraries allow high throughput screening of library compounds in either solid phase or solution phase, and position-related identification of active library compounds.

Claims

1. A method of producing a position-addressable combinatorial library of different-sequence oligomer or different-substitiuent small molecule compounds, comprising the steps of:

(i) identifying, in an array of capillary tubes, a selected subset of tubes into which a selected one of a plurality of different chemical reagents in a reagent solution is to be introduced, and a complementary subset of remaining tubes in the array,

(ii) selectively plugging a tube end of each capillary tube in said complementary subset,

(iii) placing ends of the tubes in said array in contact with said solution, under conditions effective to draw solution into the unplugged tubes only,

(iv) allowing the solution in the tubes in the selected subset to react with a wall portion of each tube on which a library compound is being formed,

(v) unplugging the plugged tubes, and

(vi) repeating steps (i)-(v) for different subsets of tubes and reagent solutions until said library is produced in said tubes.

2. The method of claim 1, wherein said placing is effective to draw such solution into the unplugged tubes by capillarity.

3. The method of claim 1, wherein said array is a two-dimensional array containing at least 500 tubes.

4. The method of claim 2, wherein said tubes have inner diameters no greater than about 50 μ

m.

5. The method of claim 1, wherein said plugging includes (i) drawing into one end region of the tubes in said array, a polymerizable solution that can be applied as a liquid and converted to a solid polymer plug by addition of a polymer initiator (ii) selectively injecting polymer initiator into the tubes in the complementary subset, thereby to form polymer plugs in the tubes in the complementary subset, and (iii) removing non-polymerized material from tubes in the selected subset.

6. The method of claim 5, wherein said injecting is carried out by an ink jet printer head.

7. The method of claim 5, wherein said polymerizable solution is an acrylamide/bisacrylamide solution, and said injecting includes selectively injecting a persulfate initiator solution into the tubes in said complementary subset of tubes.

8. The method of claim 4, wherein said plugging includes (i) drawing an radiation-polymerizable solution into one end region of the tubes in said array, (ii) selectively irradiating tubes in the complementary subset, thereby to form polymer plugs in the tubes in the complementary subset, and (iii) removing non-polymerized material from tubes in the selected subset.

9. The method of claim 1, for use in forming a combinatorial library of different-sequence oligonucleotides or oligopeptides, wherein, at each subunit addition step, the array is divided into four separate selected subsets, each of whose tubes are to receive one of four different protected nucleotides, for oligonucleotide synthesis, or are divided into N separate selected subsets, each of whose tubes are to receive one of N different protected amino acids, for oligopeptide synthesis.

10. Apparatus for producing a position-addressable combinatorial library of different-sequence oligomer or different-substituent small molecule compounds in an array of capillary tubes, comprising

(i) means for selectively plugging each tube end in a subset of tubes which is complementary to a selected subset of tubes in said array,

(ii) means for drawing a selected reagent solution into unplugged tubes in the selected subset of tubes in the array, thereby to effect a compound-addition reaction directly on the inner surface of tubes in the selected subset;

(iii) means for removing the reagent solution from the tubes in the selected subset after said reaction, and for unplugging the tubes in the complementary subset, and

(iv) means for repeating the steps performed by elements (i)-(iii) until a desired library of compounds, one library compound in each tube, is produced.

11. The apparatus of claim 10, wherein said introducing means includes (i) means for applying a polymerizable solution to end regions of all of the tubes in the array, (ii) means for selectively polymerizing the tubes in the array which form a complementary subset with the selected tubes of the array, to plug all of the array tubes except those in the selected subset, and (iii) means for introducing reagent solution by capillarity into the unplugged tubes in the array.

12. The apparatus of claim 11, wherein said polymerizing means includes an ink-jet printer head which is effective to inject a polymerization initiator solution into the tubes in the complementary subset of tubes in the array.