Functional molecule, functional molecule synthesizing amidite and target substance analysis method

• US 8,940,879 B2
• Filed: 08/30/2012
• Issued: 01/27/2015
• Est. Priority Date: 03/16/2007
• Status: Active Grant

• Current Assignee: Apta Biosciences Ltd.
• Original Assignee: Apta Biosciences Ltd.
• Inventors: Fujihara, Tsuyoshi, Fujita, Shozo
• Primary Examiner(s): Riley, Jezia
• Application Number:
  US13/599,552
• Publication Number:
  US 20130017973A1
• Time in Patent Office: 880 Days
• Field of Search: 536/4.1, 435/6.1
• US Class Current: 536/4.1
• 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 synthesizing amidite expressed by General Formula (I) below and used for the manufacture of a functional molecule, where X represents a base, Y represents a substituent and Q represents one of a hydrogen atom and a hydroxyl group;

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;

or 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,

wherein the substituent Y can be removed by at least one method selected from the group consisting of a method involving an ammonia treatment, a method involving an alkali treatment using NaOH, a method involving a treatment using hydrofluoric acid and fluoride, a method involving a treatment using hydrazine, and a method involving irradiation of light, and

wherein the functional molecule is convertible to be a molecule having a structure similar to that of naturally occurring nucleic acid by removing the substituent Y from the functional molecule;

2. The functional molecule synthesizing amidite according to claim 1, wherein the substituent Y can be removed by at least one method selected from the group consisting of a method involving an ammonia treatment, a method involving an alkali treatment using NaOH, a method involving a treatment using hydrofluoric acid and fluoride, a method involving a treatment using hydrazine, and a method involving irradiation of light, but cannot be removed by DBU treatment in acetonitrile.

3. The functional molecule synthesizing amidite according to claim 1, wherein the amidite is expressed by General Formula (II) below in which the substituent Y is additionally protected by a protection group Z, where X represents a base, Y represents a substituent, Z represents a protection group and Q represents one of a hydrogen atom and hydroxyl group;

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;

or 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,

wherein the substituent Y can be removed by at least one method selected from the group consisting of a method involving an ammonia treatment, a method involving an alkali treatment using NaOH, a method involving a treatment using hydrofluoric acid and fluoride, a method involving a treatment using hydrazine, and a method involving irradiation of light, and

wherein the functional molecule is convertible to be a molecule having a structure similar to that of naturally occurring nucleic acid by removing the substituent Y from the functional molecule;

4. The functional molecule synthesizing amidite according to claim 3, wherein the protection group Z can be removed under conditions where the substituent Y is not removed.

5. The functional molecule synthesizing amidite according to claim 3, wherein the protection group Z can be removed by DBU treatment in acetonitrile.