Functional molecule and manufacturing method therefor

• US 9,512,171 B2
• Filed: 01/19/2011
• Issued: 12/06/2016
• Est. Priority Date: 03/28/2006
• Status: Active Grant

• Current Assignee: Apta Biosciences Ltd.
• Original Assignee: Apta Biosciences Ltd.
• Inventors: Fujihara, Tsuyoshi, Kichise, Tomoyasu, Fujita, Shozo
• Primary Examiner(s): Gross, Christopher M
• Application Number:
  US12/929,359
• Publication Number:
  US 20110151510A1
• Time in Patent Office: 2,148 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: C07K 1/04 on carriers C07K1/003, C07K1/006 take precedence
• CPC Class Codes: C07K 1/1077 by covalent attachment of residues other than amino acids or peptide residues, e.g. sugars, polyols, fatty acids
• CPC Class Codes: C07K 1/22 Affinity chromatography or related techniques based upon selective absorption processes

Abstract

A novel method for efficiently manufacturing a functional molecule with affinity for a target substance is provided along with a functional molecule manufactured by this method. The method for manufacturing a functional molecule includes binding, to a resin, a specific substance having a cleavable linker which is a site that is cleavable under a specific condition to prepare a specific-substance-bound resin, treating this specific-substance-bound resin with a solution comprising functional molecule candidates, and then cleaving the cleavable linker under the specific condition to select a functional molecule which has interacted specifically with the specific substance.

Claims

1. A method for manufacturing a functional molecule, comprising:

taking a solution comprising functional molecule candidates wherein said functional molecule candidate is a modified nucleotide n-mer (n>

=2) comprising a modified nucleoside having a substituent group, and wherein said functional molecule can be transformed into a nucleic acid when said substituent group is removed by alkali treatment;

binding, to a resin, a specific substance having a cleavable linker which is a site that is cleavable under a specific condition to prepare a specific-substance-bound resin;

treating said specific-substance-bound resin with said solution comprising said functional molecule candidates;

cleaving said cleavable linker under said specific condition and selecting a functional molecule which has interacted specifically with said specific substance;

removing said substituent group of said functional molecule by alkali treatment to produce a nucleic acid; and

identifying said nucleic acid, thereby identifying said functional molecule wherein said functional molecule candidate is a modified nucleotide n-mer (n>

=2) comprising a modified nucleoside having the substituent group attached to a nucleotide base.

2. The manufacturing method according to claim 1, wherein said cleavable linker is cleavable at 50°

C. or less and in the pH range of 6.0 to 9.0.

3. The manufacturing method according to claim 1, wherein the substituent group is attached to a pyrimidine 5-position, a purine 7-position or a purine 8-position.

4. The manufacturing method according to claim 3, wherein the substituent group is attached to a pyrimidine 5-position.

5. The manufacturing method according to claim 3, wherein the substituent group is attached to a purine 7-position.

6. The manufacturing method according to claim 1, wherein said cleavable linker has disulfide bonds, vicinal diol bonds or is cleavable by light.

7. The manufacturing method according to claim 6, wherein said cleavable linker has disulfide bonds.

8. The manufacturing method according to claim 1, wherein said cleavable linker is a nucleic acid sequence cleavable with a restriction enzyme or an amino acid sequence cleavable by an enzyme.

9. The manufacturing method according to claim 1 wherein n is 10 to 50.