Sugar chain-capturing substance and use thereof

Patent metadata

Field	Value
Publication number	US-9340651-B2
Application number	US-30994507-A
Country	US
Kind code	B2
Filing date	Aug 6, 2007
Priority date	Aug 9, 2006
Publication date	May 17, 2016
Grant date	May 17, 2016

Abstract

The present invention provides a method for preparing a sample characterized by binding a substance A containing a hydrazide group to a sugar chain and/or a sugar derivative via hydrazone formation between the hydrazide group of the substance A and the reducing end of the sugar chain and/or the sugar derivative thereby to enable the separation and purification of the sugar chain and/or the sugar derivative for an analytical sample from a biological sample containing the sugar chain and/or the sugar derivative by a simple operation.

First claim

The invention claimed is:

1. A method for preparing a sample in which polymer beads A containing a hydrazine group and having a crosslinked polymer structure represented by the following formula (4) is bonded to a sugar chain and/or a sugar derivative via hydrazone formation between the hydrazine group of the polymer beads A and the reducing end of the sugar chain and/or the sugar derivative,

wherein, in the formula, R 1 and R 2 represent a hydrocarbon chain having 1 to 20 carbon atoms which may be interrupted with —O—, —S—, —NH—, —CO— or —CONH—; R 3 , R 4 and R 5 represent H, CH 3 or a hydrocarbon chain having 2 to 5 carbon atoms; and m and n represent the number of monomer units and wherein said polymer beads A having a hydrazine group and said hydrazine group has a thy weight of not less than 100 nmol per 1 mg.

2. The method for preparing a sample as set forth in claim 1 , in which said polymer beads A has a erosslinked polymer structure represented by the following formula (5),

wherein, in the formula, m and n represent the number of monomer units.

3. The method for preparing a sample as set forth in claim 1 , in which said polymer beads A is a polymer particle having an average particle diameter of equal to or more than 0.1 and equal to or less than 500 μm.

4. The method for preparing a sample as set forth in claim 1 , in which said polymer beads A is stable at the pH of 3 to 8.

5. The method for preparing a sample as set forth in claim 1 , in which said polymer beads A is stable under pressure of at least not more than 1 MPa.

6. A method for preparing a sample which involves the sugar chain capture step comprising binding polymer beads A to a sugar chain and/or a sugar derivative according to the method for preparing a sample as set forth in claim 1 , and
the sugar chain release step comprising the action of a substance B containing an aminooxy group or a hydrazide group on a complex of the polymer beads A and the sugar chain and/or the sugar derivative, captured in the sugar chain capture step, and binding said sugar chain and/or said sugar derivative while cut off from said polymer heads A to said substance B in accordance with the hydrazone-oxime exchange reaction or hydrazone-hydrazone exchange reaction occurred between said complex and said substance B.

7. The method for preparing a sample as set forth in claim 6 , in which said substance B comprises a moiety containing chromophore or fluorophore.

8. The method for preparing a sample as set forth in claim 6 , in which said substance B is a substance selected from the group consisting of the following substances containing a hydrazide group 5-Dimethylaminonaphthalene-1-sulfonyl hydrazine (Dansylhydrazine); 2-hydrazinopyridine; 9-fluorenylmethyl carbazate (Fmoc hydrazine); benzylhydrazine; 4, 4-dffluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-propionoc acid, hydrazide; 2-(6,8-difluoro-7-hydroxy-4-methylcoumarin)acetohydrazide; 7-diethylaminocoumarin-3-carboxylic acid, hydrazide (DCCH); phenylhydrazine; 1-Naphthaleneacethydrazide; 2-hydrazinobenzoic acid; and phenylacetic hydrazide; or the following substances containing an aminooxy group or a salt, of O-benzylhydroxylamine; O-phenylhydroxylamine; O-(2,3,4,5,6-pentafluorobenzl)hydroxylamine; O-(4-nitrobenzyl)hydroxylamine; 2-aminooxypyridine; 2-aminooxymethylpyridine; 4-[(aminooxyacetyl)amino]benzoic acid methyl ester; 4-[(aminooxyacetyl)amino]benzoic acid ethyl ester; and 4-[(aminooxyacety)amino]benzoic acid n-butyl ester.

9. The method for preparing a sample as set forth in claim 6 , in which said substance B comprises a moiety consisting of at least one of an arginine residue, a tryptophan residue, a phenylalanine residue, a tyrosine residue, a cysteine residue and a derivative thereof.

10. The method for preparing a sample as set forth in claim 6 , in which said substance B has a structure represented by the following formula (7),

wherein, in the structural formula, R represents -CH 3 or -CD 3 .

11. A method for preparing a sample in which a hydrazone bond between polymer beads A and a sugar chain and/or a sugar derivative is dissociated and the sugar chain and/or sugar derivative released by treating the polymer heads A under acidic conditions to, the polymer beads A having a structure represented by the following formula (4),

wherein, in the formula, R 1 and R 2 represent a hydrocarbon chain having 1 to 20 carbon atoms which may he interrupted with —O—, —S—, —NH—, —CO— or —CONH—; R 3 , R 4 and R 5 represent H, CH 3 or a hydrocarbon chain having 2 to 5 carbon atoms; and m and n represent the number of monomer units, and
wherein the polymer beads A have a hydrazine group of a dry weight of not less than 100 nmol per 1 mg.

12. The method for preparing a sample as set forth in claim 1 , in which in the formula, R 3 , R 4 and R 5 represent CH 3 or a hydrocarbon chain having 2 to 5 carbon atoms.

13. The method for preparing a sample as set forth in claim 11 , in which said polymer beads A has a crosslinked polymer structure represented by the following formula (5),

wherein, in the formula, m and n represent the number of monomer units.

Assignees

• Shimaoka Hideyuki
• Nishimura Shinichiro
• Shinohara Yasuro
• Miura Yoshiaki
• Furukawa Jun-Ichi
• Sumitomo Bakelite Co
• Univ Hokkaido Nat Univ Corp

Inventors

• Shimaoka Hideyuki
• Nishimura Shinichiro
• Shinohara Yasuro
• Miura Yoshiaki
• Furukawa Jun-Ichi

Classifications

• G01N2030/027

  Liquid chromatography · CPC title

• C08F8/30

  Introducing nitrogen atoms or nitrogen-containing groups · CPC title

• Y02P20/55

  Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups · CPC title

• G01N33/5308

  for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites · CPC title

• Y10T428/2982

  Particulate matter [e.g., sphere, flake, etc.] · CPC title