Focused acoustic energy in the preparation of peptide arrays

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.

We claim:  
     
         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.  
     
     
         7 . The method of  claim 1 , wherein each peptidic molecule is different.  
     
     
         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.  
     
     
         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.  
     
     
         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.  
     
     
         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.  
     
     
         38 . A peptide array comprised of a plurality of peptidic molecules each attached through an optional linking moiety to a substrate surface, wherein substantially none of the peptidic molecules exhibits signs of shear stress and substantially all of the peptidic molecules are intact and attached to a predetermined site on the substrate surface.  
     
     
         39 . The peptide array of  claim 38 , wherein the attachment of the peptidic molecules to the substrate surface is covalent.  
     
     
         40 . The peptide array of  claim 38 , wherein the attachment of the peptidic molecules to the substrate surface is noncovalent.  
     
     
         41 . The peptide array of  claim 38 , wherein the peptidic molecules are comprised of a plurality of peptidic molecule groups, wherein (a) all peptidic molecules within any one group are different, and (b) each peptidic molecule group is identical to each other peptidic molecule group.  
     
     
         42 . The peptide array of  claim 38 , wherein each peptidic molecule is different.  
     
     
         43 . The peptide array of  claim 38 , wherein each peptidic molecule is comprised of about 5 to about 10,000 amino acids.  
     
     
         44 . The peptide array of  claim 43 , wherein each peptidic molecule is comprised of about 5 to about 1,000 amino acids.  
     
     
         45 . The peptide array of  claim 38 , wherein each peptidic molecule is an oligopeptide, a polypeptide or protein.  
     
     
         46 . The peptide array of  claim 45 , wherein the peptidic molecule is selected from the group consisting of enzymes, antibodies, antigens, coagulation modulators, cytokines, endorphins, peptidyl hormones and kinins.  
     
     
         47 . The peptide array of  claim 38 , wherein the substrate surface is comprised of a porous material.  
     
     
         48 . The peptide array of  claim 47 , wherein the porous material is a permeable material.  
     
     
         49 . The peptide array of  claim 38 , wherein the peptidic molecules are present in a density in the range of approximately 10 to approximately 250,000 peptidic molecules per square centimeter of substrate surface.  
     
     
         50 . The peptide array of  claim 38 , wherein the peptidic molecules are present in a density of at least about 1,000,000 peptidic molecules per square centimeter of substrate surface.  
     
     
         51 . The peptide array of  claim 50 , wherein the peptidic molecules are present in a density of at least about 1,500,000 peptidic molecules per square centimeter of substrate surface.  
     
     
         52 . The peptide array of  claim 38 , wherein at least one designated site on the substrate surface includes a lipidic material.  
     
     
         53 . The peptide array of  claim 52 , wherein the lipidic material is a phospholipid.  
     
     
         54 . The peptide array of  claim 52 , wherein at least one of the peptidic molecules within the array is in a lipidic material, and at least one other of the peptidic molecules within the array is in an aqueous fluid.  
     
     
         55 . A peptide array comprised of a plurality of peptidic molecules each attached through an optional linking moiety to a predetermined site on a porous substrate surface at a density of greater than 1,000,000 peptidic molecules per square centimeter of the substrate surface.  
     
     
         56 . The peptide array of  claim 55 , wherein the density is greater than 1,500,000 peptidic molecules per square centimeter of the substrate surface.  
     
     
         57 . The peptide array of  claim 55 , wherein the porous substrate surface is a surface of a nonporous substrate.  
     
     
         58 . The peptide array of  claim 55 , wherein the porous substrate surface is a surface of a porous substrate.  
     
     
         59 . The peptide array of  claim 55 , wherein the porous substrate surface is a permeable surface.  
     
     
         60 . The peptide array of  claim 59 , wherein the permeable surface is a surface of a permeable substrate.