FUNCTIONAL MOLECULE, FUNCTIONAL MOLECULE SYNTHESIZING AMIDITE AND TARGET SUBSTANCE ANALYSIS METHOD

• US 20090053710A1
• Filed: 03/17/2008
• Published: 02/26/2009
• Est. Priority Date: 03/16/2007
• Status: Active Grant

• Current Assignee: Apta Biosciences Ltd.
• Original Assignee: Fujitsu Limited
• Inventors: FUJITA, Shozo, FUJIHARA, Tsuyoshi
• Granted Patent: US 8,288,516 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C07H 21/04 with deoxyribosyl as saccharide radical
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence

Abstract

To provide a functional molecule including a modified nucleotide unit having a substituent introduced to a base thereof, wherein the substituent is removably introduced to the base; a functional molecule synthesizing amidite that has a substituent removably introduced to its base and that is used for the manufacture of the functional molecule; and a target substance analysis method including: preparing a random pool of functional molecules using a functional molecule synthesizing amidite; screening a functional molecule having affinity for a target substance from the random pool; amplifying the functional molecules having affinity for the target substance, wherein the method further comprises, prior to the amplification step, removing a substituent from the functional molecule having affinity for the target substance.

Claims

1. A functional molecule comprising:

a modified nucleotide unit having a substituent introduced to a base thereof,

wherein the substituent is removably introduced to the base.

2. The functional molecule according to claim 1, wherein the substituent is introduced to the base through any of an aromatic carboxylic acid structure and an aliphatic carboxylic acid structure in the substituent.

3. The functional molecule according to claim 1, wherein the substituent is introduced to the base through an aromatic carboxylic acid structure in the substituent, and the base is one of adenine and cytosine.

4. The functional molecule according to claim 1, wherein the substituent is introduced to the base through an aliphatic carboxylic acid structure in the substituent, and the base is guanine.

5. The functional molecule according to claim 1, wherein the substituent can be removed by ammonia treatment.

6. The functional molecule according to claim 1, wherein the substituent can be removed by ammonia treatment but cannot be removed by DBU treatment in acetonitrile.

7. The functional molecule according to claim 1, wherein the functional molecule comprises two or more different modified nucleotide units having different substituents.

8. A functional molecule synthesizing amidite expressed by General Formula (I) below and used for the manufacture of the functional molecule according to claim 1,

wherein substituent Y is removably introduced to base X:

9. The functional molecule synthesizing amidite according to claim 8, wherein the substituent Y is introduced to the base X through any of an aromatic carboxylic acid structure and an aliphatic carboxylic acid structure in the substituent Y.

10. The functional molecule synthesizing amidite according to claim 8, wherein the substituent Y is introduced to the base X through an aromatic carboxylic acid structure in the substituent Y, and the base X is one of adenine and cytosine.

11. The functional molecule synthesizing amidite according to claim 8, wherein the substituent Y is introduced to the base X through an aliphatic carboxylic acid structure in the substituent Y, and the base X is guanine.

12. The functional molecule synthesizing amidite according to claim 8, wherein the substituent Y can be removed by ammonia treatment.

13. The functional molecule synthesizing amidite according to claim 8, wherein the substituent Y can be removed by ammonia treatment but cannot be removed by DBU treatment in acetonitrile.

14. The functional molecule synthesizing amidite according to claim 8, wherein the amidite is expressed by General Formula (II) below in which the substituent Y is additionally protected by a protection group Z:

15. The functional molecule synthesizing amidite according to claim 14, wherein the protection group Z can be removed under conditions where the substituent Y is not removed.

16. The functional molecule synthesizing amidite according to claim 14, wherein the protection group Z can be removed by DBU treatment in acetonitrile.

17. A target substance analysis method comprising:

preparing a random pool of functional molecules using a functional molecule synthesizing amidite;

screening a functional molecule having affinity for a target substance from the random pool;

amplifying the functional molecules having affinity for the target substance,

wherein the method further comprises, prior to the amplification step, removing a substituent from the functional molecule having affinity for the target substance.

18. The method according to claim 17, wherein the protection group is removed under conditions where the substituent of the functional molecule synthesizing amidite is not removed upon synthesizing of the functional molecule using the functional molecule synthesizing amidite.