Electrospraying for mass fabrication of chips and libraries

• US 6,350,609 B1
• Filed: 05/08/2000
• Issued: 02/26/2002
• Est. Priority Date: 06/20/1997
• Status: Expired due to Term

• Current Assignee: New York University
• Original Assignee: New York University
• Inventors: Morozova, Tamara Ya., Morozov, Victor
• Primary Examiner(s): Zitomer, Stephanie W.
• Assistant Examiner(s): FORMAN, BETTY J
• Application Number:
  US09/446,188
• Time in Patent Office: 659 Days
• Field of Search: 422/68.1, 422/50, 435/283.1, 435/6, 435/287.2, 427/2.13, 205/158, 436/86
• US Class Current: 506/16
• 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 2219/00371 comprising electrodes
• CPC Class Codes: B01J 2219/0043 for direct application of reagents, e.g. through openings in a shutter
• CPC Class Codes: B01J 2219/00432 Photolithographic masks
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00585 Parallel 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 2219/00626 Covalent
• CPC Class Codes: B01J 2219/00628 Ionic
• CPC Class Codes: B01J 2219/0063 Other, e.g. van der Waals forces, hydrogen bonding
• 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 2219/00677 Ex-situ synthesis followed by deposition on the substrate
• CPC Class Codes: B01J 2219/00702 Processes involving means for analysing and characterising the products
• CPC Class Codes: B01J 2219/00711 : Light-directed synthesis
• CPC Class Codes: B01J 2219/00713 : Electrochemical synthesis
• CPC Class Codes: B01J 2219/00725 : Peptides
• CPC Class Codes: B05B 12/36 : Side shields, i.e. shields extending in a direction substantially parallel to the spray jet
• CPC Class Codes: B05B 5/002 : comprising means for neutralising the spray of charged droplets or particules
• CPC Class Codes: B05B 5/0255 : spraying and depositing by electrostatic forces only
• CPC Class Codes: B82Y 30/00 : Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C40B 40/10 : Libraries containing peptides or polypeptides, or derivatives thereof
• CPC Class Codes: C40B 60/14 : for creating libraries
• CPC Class Codes: G01N 33/543 : with an insoluble carrier for immobilising immunochemicals
• CPC Class Codes: G01N 33/545 : Synthetic resin

Abstract

A method of fabricating deposits of non-volatile substances, including biomacromolecules, in the form of spots and filing on a substrate surface by electrospray, where the deposits are used to determine the interaction of the deposited non-volatile substances to other substances. Also included in this method is the mass fabrication on a single chip of an array of single and multicomponent microsamples.

Claims

1. A method for fabricating a deposit of a non-volatile biologically functional and/or biologically active substance on a substrate, comprising:

electrospraying a solution of said non-volatile biologically functional and/or biologically active substance under a voltage effective to atomize said solution without destroying the functional properties of the non-volatile biologically functional and/or biologically active substance; and

depositing the electrosprayed substance in the form of dry deposit on contact with a surface of the substrate as the biologically functional and/or biologically active substance.

2. The method in accordance with claim 1, wherein during said electrospraying and depositing steps, electrostatic charge on the electrosprayed substance deposited on the surface of the substrate is neutralized.

3. The method in accordance with claim 2, wherein the substrate is made of an electrically conductive material, and the electrostatic charge on the electrospray products are neutralized by a local increase in conductivity of a dielectric layer placed on the substrate.

4. The method in accordance with claim 3, wherein the dielectric layer is made of a photo-conductive material and the local increase in conductivity of the dielectric layer is performed by locally irradiating the photo-conductive insulating layer with light.

5. The method in accordance with claim 3, wherein the dielectric layer is made locally conductive by injection of charge carriers from an external source and the local increase in conductivity of the dielectric layer is performed by locally injecting charge carriers into the dielectric layer.

6. The method in accordance with claim 3, wherein the dielectric layer is made of a thermally conductive material, and the local increase in conductivity of the dielectric layer is performed by locally heating the dielectric layer.

7. The method in accordance with claim 3, wherein the local increase in conductivity is made by perforating the dielectric layer.

8. The method in accordance with claim 3, wherein the size of a deposit of the non-volatile biologically functional and/or biologically active substance on the surface of the substrate is smaller than the size of an area in which conductivity is locally increased.

9. The method in accordance with claim 3, wherein a plurality of confined electrostatic fields are created on the substrate to deposit simultaneously a plurality of non-volatile biologically functional and/or biologically active substances on the substrate.

10. The method in accordance with claim 9, wherein the position of said plurality of confined electrostatic fields is shifted and said electrospraying and depositing steps are repeated.

11. The method in accordance with claim 10, wherein a solution comprising a different non-volatile biologically functional and/or biologically active substance is deposited in a repetition of said electrospraying and depositing steps.

12. The method in accordance with claim 3, wherein a collecting ring is provided around the substrate area with increased conductivity.

13. The method in accordance with claim 12, wherein a potential is applied to the collecting ring so as to achieve stable electrospray with a maximum fraction of the total current from a capillary passing through the substrate.

14. The method in accordance with claim 2, wherein said depositing step is conducted such that a deposit of said biologically functional and/or biologically active substance is formed at a predetermined position on the substrate, and in a predetermined form and shape under control of an electrostatic field.

15. The method in accordance with claim 2, wherein the deposited non-volatile biologically functional and/or biologically active substance forms a film.

16. The method in accordance with claim 15, wherein said film of the deposited non-volatile biologically functional and/or biologically active substance has a non-round shape.

17. The method in accordance with claim 15, further comprising a step of applying a conductive or semiconductive layer onto a substrate before electrospraying, whereby the conductive or semiconductive layer provides a surface for depositing the film by electrospray.

18. The method in accordance with claim 17, further comprising a step of detaching the film of the non-volatile biologically functional and/or biologically active substance from the conductive or semiconductive layer.

19. The method in accordance with claim 18, wherein the conductive layer contains a chemically or physically solubilized material.

20. The method in accordance with claim 19, wherein the conductive layer contains hydrophilic polymers.

21. The method in accordance with claim 15, wherein the surface on which the electrosprayed film is deposited is a surface of a substrate having low conductivity.

22. The method in accordance with claim 21, wherein the surface on which the electrosprayed film is deposited is porous.

23. The method in accordance with claim 1, wherein a dielectric protective screen is disposed around a capillary to surround the electrosprayed products and prevent scatter.

24. The method in accordance with claim 23, wherein said dielectric protective screen is perforated.

26. The method in accordance with claim 1, wherein the non-volatile biologically functional and/or biologically active substance is a biomacromolecule.

27. The method in accordance with claim 26, further comprising the step of cross-linking the sample of deposited biomacromolecule.

28. The method in accordance with claim 27, wherein the biomacromolecule is a protein.

29. The method in accordance with claim 28, wherein the electrosprayed solution further comprises a soluble protein stabilizer.

30. The method in accordance with claim 29, further comprising the step of washing away the protein stabilizer from the cross-linked protein film to obtain a porous protein film.

31. The method in accordance with claim 28, further comprising the step of contacting the deposited protein film with a reagent to produce a homogeneous film.

32. The method in accordance with claim 31, wherein the reagent is water vapor.

33. The method in accordance with claim 28, wherein the protein is deposited as nanoclusters of protein molecules with a size of the cluster between 5 and 50 nm.

34. The method in accordance with claim 33, wherein the step of electrospraying is conducted in an atmosphere in the range of about 10% to 40% humidity.

35. A protein film produced by the method according to claim 33, wherein the deposited nanoclusters of protein molecules are densely packed but not completely coalesced and said film has intercluster channels penetrable by ligands.

36. The method in accordance with claim 26, wherein the biomacromolecule is a nucleic acid molecule.

37. The method in accordance with claim 1, wherein the solution containing the non-volatile biologically functional and/or biologically active substance is electrosprayed from a capillary, which capillary is set to a potential to effect sufficient electrospray without resulting in a corona discharge.

38. The method in accordance with claim 37, wherein a current in a range of about 1-50 nA is maintained through the capillary.

39. The method in accordance with claim 1, wherein the solution containing a non-volatile biologically functional and/or biologically active substance has a conductivity less than about 500 microSiemens per cm.

40. The method in accordance with claim 1, further comprising a step of preventing intermolecular reaction between deposited electrospray products or between deposited electrospray products and molecules present on the substrate.

41. The method in accordance with claim 1, wherein said electrospraying is performed with capillaries.

42. The method in accordance with claim 1, wherein said voltage effective to atomize said solution without destroying the functional properties of the non-volatile biologically functional and/or biologically active substance is less than 7-8 kV at a current less than 500-1500 nA.

25. The method in accordance with 1, wherein the electrosprayed solution further comprises a stabilizer.

43. A method for fabricating a deposit of a non-volatile biologically functional and/or biologically active substance on a substrate, comprising:

electrospraying a solution of said non-volatile biologically functional and/or biologically active substance under conditions which prevent corona discharge with partial or complete evaporation of solvent; and

depositing the electrosprayed biologically functional and/or biologically active substance on a surface of the substrate as a dry charged residue or as a highly concentrated solution.