Methods for synthesis of an oligopeptide microarray
An oligopeptide microarray and methods for the synthesis thereof are presented. Further presented is a microarray on a solid support comprising at least about 10,000 oligopeptide features per cm 2 and preferably at least about 50,000 oligopeptide features per cm 2 .
1. A method for synthesizing an oligopeptide microarray, wherein the method comprises the steps of:
a) coupling to a reactive amino group attached directly or indirectly to a surface of a plastic solid support an amino acid comprising an amino group protected by a photolabile moiety selected from 2-(2-nitrophenyl)-propoxycarbonyl (NPPOC) and a derivative thereof, with the proviso that the photolabile moiety is 2-(2-nitro-4-benzoylphenyl)-2′-propyl-1′-oxycarbonyl (benzoyl-NPPOC) when the amino acid is tyrosine wherein the method includes coupling at least one tyrosine;
b) optionally capping unreacted amino acids;
c) optionally washing the plastic solid support;
d) photoirradiating the protected amino group such that the photoirradiated amino group is reactive; and
e) repeating steps a) to d) for a predetermined number of times, whereby an oligopeptide is synthesized.
2. The method according to claim 1 , wherein photoirradiating is spatially resolved.
3. The method according to claim 1 , wherein the surface comprises ε-amino-hexanoic-acid.
4. The method according to claim 1 , wherein the plastic solid support has at least 50% light transmission when photoirradiating is performed between 350 and 410 nm.
5. The method according to claim 1 , wherein photoirradiating is performed between 350 and 410 nm.
6. The method according to claim 5 , wherein photoirradiating is performed between 350 and 375 nm, between 360 and 370 nm, between 363 and 367 nm, or at 365 nm.
7. The method according to claim 1 , wherein synthesizing comprises maskless photolithography.
8. The method according to claim 7 , wherein maskless photolithography comprises using a digital micromirror device.
9. The method according to claim 1 , wherein the amino acid is selected from the group consisting of tyrosine, histidine, alanine, valine, glycine, leucine, isoleucine, aspartic acid, glutamic acid, serine, glutamine, asparagine, threonine, arginine, proline, phenylalanine, tryptophan, cysteine, methionine, and lysine.
10. The method according to claim 1 , wherein the derivative of NPPOC is benzoyl-NPPOC.
11. The method according to claim 1 , wherein the predetermined number of times is between 6 and 24, whereby the oligopeptide comprises 6 to 24 amino acids.
12. The method according to claim 11 , wherein the predetermined number of times is between 9 and 18, whereby the oligopeptide comprises 9 to 18 amino acids.
13. The method according to claim 1 , wherein photoirradiating is performed in the presence of a polar organic solvent selected from the group consisting of dimethylsulfoxide, N-methyl-2-pyrrolidone, dimethylformamide, acetonitrile, methanol, ethanol, and propanol, or a base selected from the group consisting of hydrazine, hydroxylamine, and imidazole.
14. The method according to claim 1 , wherein the step of photoirradiating occurs over a time period of less than about 5 minutes.
15. The method according to claim 1 , further comprising positioning photoirradiation beams on the solid support prior to at least every 4th photoirradiating step.
16. The method according to claim 1 , further comprising adjusting the position of the solid support.
17. The method according to claim 1 , further comprising synthesizing a plurality of oligopeptide features on the oligopeptide microarray by performing each of the steps a) to e), wherein the oligopeptide microarray comprises at least 10,000 of the oligopeptide features per cm 2 .
18. A method for synthesizing an oligopeptide microarray, wherein the method comprises the steps of:
a) coupling to a reactive amino group attached directly or indirectly to a surface of a solid support an amino acid comprising an amino group protected by a photolabile moiety selected from 2-(2-nitrophenyl)-propoxycarbonyl (NPPOC) and a derivative thereof, with the proviso that the photolabile moiety is 2-(2-nitro-4-benzoylphenyl)-2′-propyl-1′-oxycarbonyl (benzoyl-NPPOC) when the amino acid is tyrosine wherein the method includes coupling at least one tyrosine;
b) optionally capping unreacted amino acids;
c) optionally washing the solid support;
d) photoirradiating the protected amino group such that the photoirradiated amino group is reactive; and
e) repeating steps a) to d) for a predetermined number of times, whereby an oligopeptide is synthesized.
19. The method according to claim 18 , wherein photoirradiating is spatially resolved.
20. The method according to claim 18 , wherein the surface comprises ε-amino-hexanoic-acid.
21. The method according to claim 18 , wherein the solid support has at least 50% light transmission when photoirradiating is performed between 350 and 410 urn.
22. The method according to claim 18 , wherein photoirradiating is performed between 350 and 410 nm.
23. The method according to claim 22 , wherein photoirradiating is performed between 350 and 375 nm, between 360 and 370 nm, between 363 and 367 nm, or at 365 nm.
24. The method according to claim 18 , wherein synthesizing comprises maskless photolithography.
25. The method according to claim 24 , wherein maskless photolithography comprises using a digital micromirror device.
26. The method according to claim 18 , wherein the amino acid is selected from the group consisting of tyrosine, histidine, alanine, valine, glycine, leucine, isoleucine, aspartic acid, glutamic acid, serine, glutamine, asparagine, threonine, arginine, proline, phenylalanine, tryptophan, cysteine, methionine, and lysine.
27. The method according to claim 18 , wherein the derivative of NPPOC is benzoyl-NPPOC.
28. The method according to claim 18 , wherein the predetermined number of times is between 6 and 24, whereby the oligopeptide comprises 6 to 24 amino acids.
29. The method according to claim 28 , wherein the predetermined number of times is between 9 and 18, whereby the oligopeptide comprises 9 to 18 amino acids.
30. The method according to claim 18 , wherein photoirradiating is performed in the presence of a polar organic solvent selected from the group consisting of dimethylsulfoxide, N-methyl-2-pyrrolidone, dimethylformamide, acetonitrile, methanol, ethanol, and propanol, or a base selected from the group consisting of hydrazine, hydroxylamine, and imidazole.
31. The method according to claim 18 , wherein the step of photoirradiating occurs over a time period of less than about 5 minutes.
32. The method according to claim 18 , further comprising positioning photoirradiation beams on the solid support prior to at least every 4th photoirradiating step.
33. The method according to claim 18 , further comprising adjusting the position of the solid support.
34. The method according to claim 18 , further comprising synthesizing a plurality of oligopeptide features on the oligopeptide microarray by performing each of the steps a) to e), wherein the oligopeptide microarray comprises at least 10,000 oligopeptide features per cm 2 .
35. A method for synthesizing an oligopeptide microarray, wherein the method comprises the steps of:
a) coupling to a reactive amino group attached directly or indirectly to a surface of a solid support an amino acid comprising an amino group protected by a photolabile moiety, the photolabile moiety selected from 2-(2-nitrophenyl)-propoxycarbonyl (NPPOC) and a derivative thereof;
b) optionally capping unreacted amino acids;
c) optionally washing the solid support;
d) photoirradiating the protected amino group such that the photoirradiated amino group is reactive; and
e) repeating steps a) to d) for a predetermined number of times, whereby an oligopeptide is synthesized,
wherein the predetermined number of times is at least one,
wherein the method includes coupling at least one tyrosine, and at least one amino acid selected from the group consisting of histidine, alanine, valine, glycine, leucine, isoleucine, aspartic acid, glutamic acid, serine, glutamine, asparagine, threonine, arginine, proline, phenylalanine, tryptophan, cysteine, methionine, and lysine,
wherein the photolabile moiety is benzoyl-NPPOC when the amino acid is tyrosine, and
wherein the photolabile moiety is not benzoyl-NPPOC when the amino acid is selected from the group consisting of histidine, alanine, valine, glycine, leucine, isoleucine, aspartic acid, glutamic acid, serine, glutamine, asparagine, threonine, arginine, proline, phenylalanine, tryptophan, cysteine, methionine, and lysine.
36. The method according to claim 35 , wherein the step of photoirradiating occurs over a time period of less than about 5 minutes.