Direct molecular patterning using a micro-stamp gel

• US 5,948,621 A
• Filed: 09/30/1997
• Issued: 09/07/1999
• Est. Priority Date: 09/30/1997
• Status: Expired due to Fees

• Current Assignee: the united states of america as represented by the secretary of the navy
• Original Assignee: the united states of america as represented by the secretary of the navy
• Inventors: Turner, David C., Martin, Brett, Gaber, Bruce P.
• Primary Examiner(s): Wax, Robert A.
• Assistant Examiner(s): Stole, Einar
• Application Number:
  US08/940,178
• Time in Patent Office: 707 Days
• Field of Search: 427/150, 435/4, 435/5, 435/6, 435/7.1
• US Class Current: 435/6.11
• 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 2219/00382 Stamping
• CPC Class Codes: B01J 2219/00527 Sheets
• 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 2219/0063 Other, e.g. van der Waals forces, hydrogen bonding
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B41M 3/006 Patterns of chemical products used for a specific purpose, e.g. pesticides, perfumes, adhesive patterns; use of microencapsulated material; Printing on smoking articles
• CPC Class Codes: C12Q 1/68 involving nucleic acids
• CPC Class Codes: C12Q 1/6834 Enzymatic or biochemical coupling of nucleic acids to a solid phase
• CPC Class Codes: C40B 60/14 for creating libraries
• CPC Class Codes: G01N 1/30 Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
• CPC Class Codes: G01N 2650/00 Assays involving polymers whose constituent monomers bore biological functional groups before polymerization, i.e. vinyl, acryl derivatives of amino acids, sugars
• CPC Class Codes: Y10S 977/887 Nanoimprint lithography, i.e. nanostamp

Abstract

A stamp for transferring molecules and molecular patterns to a substrate face includes a backing and a polymeric gel bound to the backing and loaded with the a molecular species. Where the molecule to be patterned is a biomolecule, such as a protein or nucleic acid, the polymeric gel is typically a hydrogel. Exemplary hydrogels include sugar-based polyacrylates and polyacrylamides.

Claims

1. A molecular stamp comprising:

a solid substrate support;

a polymeric gel covalently bound to said solid support so as to form an exposed patterned surface, said patterned surface including a raised region comprising said polymeric gel and at least one indentation within said raised region;

said polymeric gel having absorbed therein a liquid vehicle, said liquid vehicle having a first molecular species dissolved or colloidally suspended therein.

2. The molecular stamp of claim 1, wherein said polymeric gel is a hydrogel.

3. The molecular stamp of claim 2, wherein said indentation is defined by an absence of said polymeric hydrogel.

4. The molecular stamp of claim 3, wherein said raised region comprises at least two distinct raised portions comprising said polymeric hydrogel, said at least two raised portions being isolated from each other by said indentation.

5. The molecular stamp of claim 2, wherein said molecular species is a biomolecule.

6. The molecular stamp of claim 5, wherein said molecular species is a protein.

7. The molecular stamp of claim 2, wherein said polymeric hydrogel is a sugar-based polyacrylate hydrogel.

8. The molecular stamp of claim 7, wherein said polymeric hydrogel is formed by crosslinking 6-acryloyl-β

-O-methylgalactoside.

9. A molecular stamp comprising an array of capillaries, each of said capillaries having an open end, a plurality of said capillaries each including therein a gel extending from within said capillaries to beyond said open end of said capillaries, said gel in at least one of said capillaries having absorbed therein a liquid vehicle having a first molecular species dissolved or colloidally dispersed therein.

10. The molecular stamp of claim 9, wherein said capillaries are in fluid connection with a reservoir including said liquid vehicle and said first molecular species dissolved or colloidally dispersed therein.

11. The molecular stamp of claim 9, wherein said molecular species is a biomolecule.

12. The molecular stamp of claim 11, wherein said molecular species is a protein.

13. The molecular stamp of claim 9, wherein said gel is a hydrogel.

14. The molecular stamp of claim 13, wherein said polymeric hydrogel is a sugar-based polyacrylate hydrogel.

15. The molecular stamp of claim 14, wherein said polymeric hydrogel is formed by crosslinking 6-acryloyl-β

-O-methylgalactoside.

16. The molecular stamp of claim 9, wherein said gel in another one of said capillaries has absorbed therein a second molecular species, said second molecular species being distinct from said first molecular species, and wherein each of said capillaries has only one molecular species dissolved or suspended in said liquid vehicle absorbed therein.

17. A method of transferring a molecular species to a substrate, said method comprising the steps of:

loading a molecular species dissolved or colloidally suspended in a liquid vehicle into a polymeric gel, said polymeric gel being covalently bound on one surface thereof to a solid support so as to form an exposed patterned surface on another surface of said gel, said patterned surface including a raised region comprising said polymeric gel and at least one indentation within said raised region;

contacting said exposed portion of said polymeric gel to a substrate, thereby transferring said loaded molecular species from said polymeric gel onto said substrate.

18. The method of claim 17, wherein, during said contacting step, said substrate is at a temperature of above 0°

C. and below about 10°

C.

19. The method of claim 17, wherein said polymeric gel is a hydrogel and said indentation is defined by an absence of said polymeric hydrogel.

20. The method of claim 19, wherein said raised region comprises at least two distinct raised portions comprising said polymeric hydrogel, said at least two raised portions being isolated from each other by said indentation, and wherein said molecular species loaded in one of said at least two raised regions is a different molecular species than the molecular species loaded in the other one of said at least two raised regions.

21. The method of claim 17, wherein said substrate is a live cell.