Peptide arrays and methods of preparation

• US 20070015213A1
• Filed: 07/13/2006
• Published: 01/18/2007
• Est. Priority Date: 09/25/2000
• Status: Active Grant

• Current Assignee: Labcyte, Inc. (Danaher Corporation)
• Original Assignee: Picoliter Inc. (Danaher Corporation)
• Inventors: Mutz, Mitchell, Ellson, Richard
• Granted Patent: US 7,901,039 B2
• Time in Patent Office: Days
• US Class Current: 435/7.100
• 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 2219/00378 Piezoelectric or ink jet dispensers
• 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 2219/00608 DNA chips
• CPC Class Codes: B01J 2219/0061 The surface being organic
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00626 Covalent
• 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 2219/00641 the porous medium being continuous, e.g. porous oxide substrates
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: B05B 17/0615 spray being produced at the free surface of the liquid or other fluent material in a container and subjected to the vibrations
• CPC Class Codes: B41J 2/14008 Structure of acoustic ink jet print heads
• 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 60/14 : for creating libraries

Abstract

The present invention relates to arrays of peptidic molecules and the preparation of peptide arrays using focused acoustic energy. The arrays are prepared by acoustically ejecting peptide-containing fluid droplets from individual reservoirs towards designated sites on a substrate for attachment thereto.

Claims

1. A method for preparing an array comprised of a plurality of peptidic molecules attached to a substrate surface, the method comprising applying focused acoustic energy to each of a plurality of reservoirs each containing a peptidic molecule in a fluid, wherein the focused acoustic energy is applied in a manner effective to eject a droplet from each reservoir toward a different site on a substrate surface.

2. The method of claim 1, wherein the focused acoustic energy is applied to each of the plurality of reservoirs by (a) acoustically coupling each reservoir in succession to an ejector that produces acoustic radiation;

and (b) following each acoustic coupling step, activating the ejector to generate acoustic radiation having a focal point sufficiently near the fluid surface so as to eject a fluid droplet from the reservoir toward a designated site on the substrate surface.

3. The method of claim 1, wherein each peptidic molecule is comprised of about 5 to about 10,000 amino acids.

4. The method of claim 3, wherein each peptidic molecule is comprised of about 5 to about 1,000 amino acids.

5. The method of claim 1, wherein each peptidic molecule is an oligopeptide, a polypeptide or protein.

6. The method of claim 5, wherein the peptidic molecule is selected from the group consisting of enzymes, antibodies, antigens, coagulation modulators, cytokines, endorphins, peptidyl hormones and kinins.

8. The method of claim 1, wherein each of the ejected droplets has a volume in the range of about 1 pL to about 5 pL.

9. The method of claim 1, wherein each of the ejected droplets has a volume of less than about 1 pL.

10. The method of claim 1, wherein the fluid has a viscosity of least about 40 cP.

11. The method of claim 10, wherein the fluid has a viscosity of at least about 100 cP.

12. The method of claim 11, wherein the fluid has a viscosity of at least about 1000 cP.

31. The method of either claim 1 or claim 13, further comprising pretreating the substrate surface with an activating agent.

32. The method of claim 31, wherein the activating agent is cyanogen bromide, tresyl chloride or N-hydroxysuccinimide.

33. The method of either claim 1 or claim 13, wherein the substrate surface is comprised of a porous material.

34. The method of claim 33, wherein the porous material is a permeable material.

7. The method of claim I, wherein each peptidic molecule is different.

13. A method for preparing a peptide array comprised of a plurality of peptidic molecules attached to a substrate surface, the method comprising:

(a) acoustically coupling a first reservoir containing a first peptidic molecule in a first fluid to an ejector that produces acoustic radiation;

(b) activating the ejector to generate acoustic radiation having a focal point sufficiently near the surface of the first fluid so as to eject a droplet thereof toward a first designated site on the substrate surface;

(c) acoustically coupling a second reservoir containing a second peptidic molecule in a second fluid to the ejector;

(d) activating the ejector as in step (b) to eject a droplet of the second fluid from the second reservoir toward a second designated site on the substrate surface; and

(e) repeating steps (c) and (d) with additional reservoirs each containing a peptidic molecule in a fluid until a droplet has been ejected from each reservoir.

14. The method of claim 13, wherein the time period between activation steps is no longer than about 1 second.

15. The method of claim 14, wherein the time period between activation steps is no longer than about 0.1 second.

16. The method of claim 15, wherein the time period between activation steps is no longer than about 0.01 second.

17. The method of claim 16, wherein the time period between activation steps is no longer than about 0.001 second.

18. The method of claim 13, wherein steps (b) and (d) result in attachment of the first and second peptidic molecules, respectively, to the first and second designated sites on the substrate surface.

19. The method of claim 18, wherein the attachment is covalent.

20. The method of claim 18, wherein the attachment is noncovalent.

21. The method of claim 13, wherein each peptidic molecule is comprised of about 5 to about 10,000 amino acids.

22. The method of claim 21, wherein each peptidic molecule is comprised of about 5 to about 1,000 amino acids.

23. The method of claim 13, wherein each peptidic molecule is an oligopeptide, a polypeptide or protein.

24. The method of claim 23, wherein the peptidic molecule is selected from the group consisting of enzymes, antibodies, antigens, coagulation modulators, cytokines, endorphins, peptidyl hormones and kinins.

25. The method of claim 13, wherein each peptidic molecule is different.

26. The method of claim 13, wherein each of the ejected droplets has a volume in the range of about 1 pL to about 5 pL.

27. The method of claim 13, wherein each of the ejected droplets has a volume of less than about 1 pL.

28. The method of claim 13, wherein each fluid has a viscosity of least about 40 cP.

29. The method of claim 28, wherein the fluid has a viscosity of at least about 100 cP.

30. The method of claim 29, wherein the fluid has a viscosity of at least about 1000 cP.

35. A method for preparing a peptide array comprised of a plurality of peptidic molecules attached to a porous substrate surface, the method comprising applying focused acoustic energy to each of a plurality of fluid-containing reservoirs each containing a peptidic molecule, wherein the focused acoustic energy is applied in a manner effective to eject a droplet having a volume of at most about 1 pL from each reservoir toward a different designated site on the porous substrate surface.

36. The method of claim 35, wherein the array is prepared at a density of at least about 1,000,000 peptidic molecules per square centimeter of the substrate surface.

37. The method of claim 35, wherein the array is prepared at a density of at least about 1,500,000 peptidic molecules per square centimeter of the substrate surface.