Peptide arrays and methods of preparation
View Patent ↗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.
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, wherein each peptidic molecule is an oligopeptide, a polypeptide or protein.
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 the peptidic molecule is selected from a group consisting of enzymes, antibodies, antigens, coagulation modulators, cytokines, endorphins, peptidyl hormones and kinins.
6. The method of claim 1 , wherein each peptidic molecule is different.
7. The method of claim 1 , wherein each ejected droplet has a volume in the range of about 1 pL to about 5 pL.
8. The method of claim 1 , wherein each ejected droplet has a volume of less than about 1 pL.
9. The method of claim 1 , wherein the fluid has a viscosity of at least about 40 cP.
10. The method of claim 9 , wherein the fluid has a viscosity of at least about 100 cP.
11. The method of claim 10 , wherein the fluid has a viscosity of at least about 1000 cP.
12. 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, wherein each peptidic molecule is an oligopeptide, a polypeptide or protein.
13. The method of claim 12 , wherein the time period between activation steps is no longer than about 1 second.
14. The method of claim 13 , wherein the time period between activation steps is no longer than about 0.1 second.
15. The method of claim 14 , wherein the time period between activation steps is no longer than about 0.01 second.
16. The method of claim 15 , wherein the time period between activation steps is no longer than about 0.001 second.
17. The method of claim 12 , 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.
18. The method of claim 17 , wherein the attachment is covalent.
19. The method of claim 17 , wherein the attachment is noncovalent.
20. The method of claim 12 , wherein each peptidic molecule is comprised of about 5 to about 10,000 amino acids.
21. The method of claim 20 , wherein each peptidic molecule is comprised of about 5 to about 1,000 amino acids.
22. The method of claim 12 , wherein each peptidic molecule is selected from a group consisting of enzymes, antibodies, antigens, coagulation modulators, cytokines, endorphins, peptidyl hormones and kinins.
23. The method of claim 12 , wherein each peptidic molecule is different.
24. The method of claim 12 , wherein each ejected droplet has a volume in the range of about 1 pL to about 5 pL.
25. The method of claim 12 , wherein each ejected droplet has a volume of less than about 1 pL.
26. The method of claim 12 , wherein each fluid has a viscosity of at least about 40 cP.
27. The method of claim 26 , wherein the fluid has a viscosity of at least about 100 cP.
28. The method of claim 27 , wherein the fluid has a viscosity of at least about 1000 cP.
29. The method of either claim 1 or claim 12 , further comprising pretreating the substrate surface with an activating agent.
30. The method of claim 29 , wherein the activating agent is cyanogen bromide, tresyl chloride or N-hydroxysuccinimide.
31. The method of either claim 1 or claim 12 , wherein the substrate surface is comprised of a porous material.
32. The method of claim 31 , wherein the porous material is a permeable material.
33. 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, wherein each peptidic molecule is an oligopeptide, a polypeptide or protein.
34. The method of claim 33 , wherein the array is prepared at a density of at least about 1,000,000 peptidic molecules per square centimeter of the substrate surface.
35. The method of claim 33 , wherein the array is prepared at a density of at least about 1,500,000 peptidic molecules per square centimeter of the substrate surface.