Photolithographic solid-phase polymer synthesis

• US 7,144,700 B1
• Filed: 07/21/2000
• Issued: 12/05/2006
• Est. Priority Date: 07/23/1999
• Status: Expired due to Term

• Current Assignee: Affymetrix, Inc. (Thermo Fisher Scientific Incorporated), University of Maryland
• Original Assignee: Affymetrix, Inc. (Thermo Fisher Scientific Incorporated)
• Inventors: Falvey, Daniel E., Fidanza, Jacqueline A., Feldman, Brian M., McGall, Glenn
• Primary Examiner(s): Paras, Jr., Peter
• Assistant Examiner(s): Tran, My-Chau T
• Application Number:
  US09/620,780
• Time in Patent Office: 2,328 Days
• Field of Search: 435/7.1, 435/7.2, 435/DIG.42, 435/DIG.49, 436/518, 530/333, 530/334, 536/25.3, 536/25.31, 536/25.32, 536/25.33, 536/25.34
• US Class Current: 435/6.11
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: Y02P 20/55 Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
• CPC Class Codes: Y10T 436/17 Nitrogen containing
• CPC Class Codes: Y10T 436/23 Carbon containing

Abstract

Methods, employing a polycyclic hydrocarbon or a polycyclic heteroaromatic compound as sensitizers, are provided to increase the efficiency of removing, by irradiation, photolabile protecting groups that mask reactive sites on synthesis intermediaries. Preferred groups of photolabile protecting moieties include: ((α-methyl-2-nitropiperonyl)-oxy)carbonyl (MeNPOC), ((Phenacyl)-oxy)carbonyl (PAOC), O-(9-phenylxanthen-9-yl) (PIXYL), and ((2-methylene-9,10-anthraquinone)-oxy)carbonyl (MAQOC). In conjunction with using the sensitizers and protecting groups described above, a method of forming, from component molecules, a plurality of compounds on a support, each compound occupying a separate predefined region of the support is provided. These resulting solid-phase arrays are useful, for example, to assay for the presence of biochemical products in biological samples.

Claims

1. A method for removing a photolabile protecting group attached by a chemical bond to at least one monomer, comprising irradiating said photolabile protecting group in the presence of at least one sensitizer;

wherein said monomer is covalently attached to a solid support or a polymer, wherein said monomer is a nucleic acid, nucleotide, or nucleoside, and wherein said sensitizer is 9,10-dimethylanthracene.

2. The method of claim 1, wherein said photolabile protecting group is selected from the group consisting of ((α

-methyl-2-nitropiperonyl)-oxy)carbonyl (MeNPOC), ((Phenacyl)-oxy)carbonyl (PAOC), and O-(9-phenylxanthen-9-yl) (PIXYL).

3. The method of claim 1, wherein the monomer is linked to the photolabile protecting group via a hydroxy or amine group.

4. The method of claim 1, wherein the nucleoside is a ribonucleoside or deoxyribonucleoside.

5. The method of claim 4, wherein the ribonucleoside is adenosine, cytidine, uracil, or guanosine.

6. The method of claim 4, wherein the deoxyribonucleoside is deoxyadenosine, deoxycytidine, deoxythymidine, or deoxyguanosine.

8. The method of claim 1, wherein irradiating is at wavelengths greater than about 340 nm.

9. The method of claim 8, wherein irradiating is at wavelengths greater than about 365 nm.

7. A method for removing a photolabile protecting group attached by a chemical bond to at least one monomer, comprising irradiating said photolabile protecting group in the presence of at least one sensitizer;

wherein said monomer is covalently attached to a solid support, wherein the monomer is a nucleotide;

the photolabile group is ((α

-methyl-2-nitropiperonyl)-oxy)carbonyl (MeNPOC); and

the sensitizer is 9,10-dimethylanthracene.