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Sugar-immobilized metal nanoparticle, method for measuring sugar-protein interaction using the same and method for recovering protein from sugar-protein interactant achieved

Foreign code F110005671
File No. V350P001WO
Posted date Sep 9, 2011
Country United States of America
Application number 92087806
Gazette No. 20090240032
Gazette No. 8067393
Date of filing May 26, 2006
Gazette Date Sep 24, 2009
Gazette Date Nov 29, 2011
International application number JP2006310592
International publication number WO2006126689
Date of international filing May 26, 2006
Date of international publication Nov 30, 2006
Priority data
  • P2005-154550 (May 26, 2005) JP
  • 2006WO-JP310592 (May 26, 2006) WO
Title Sugar-immobilized metal nanoparticle, method for measuring sugar-protein interaction using the same and method for recovering protein from sugar-protein interactant achieved
Abstract (US8067393)
It is intended to provide a stable novel sugar-immobilized metal nanoparticle capable of easily immobilizing a sugar chain, a method for measuring sugar-protein interaction easily and at a low cost using the same without labeling, and a method for simply recovering a protein from a sugar-protein interactant.
A maltose-immobilized gold nanoparticle was obtained by binding a ligand complex, in which maltose and a linker compound had been bound to each other, to a gold nanoparticle.
By adding this maltose-immobilized gold nanoparticle to a dilution series of concanavalin A, a sugar-protein interactant of maltose and ConA was formed, and red-purple color derived from a colloidal solution of maltose-immobilized gold nanoparticle disappeared.
That is, sugar-protein interaction could be confirmed by visual observation without labeling.
Scope of claims [claim1]
1. A sugar-immobilized metal nanoparticle, which comprises: a ligand complex of a structure including a linker compound comprising thioctic acid and m-phenylenediamine at a molar ratio of 1:1, and a sugar having a reducing end, the linker compound including a hydrocarbon structure which includes: a hydrocarbon inducing chain with an aromatic amino group at an end and a carbon-nitrogen bond in a backbone; and a sulfur atom, the sugar being bound to the linker compound via the amino group; and
gold bound to the ligand complex,
wherein the sugar-immobilized metal nanoparticle is obtained by mixing the ligand complex with a solution of a gold nanoparticle prepared by mixing sodium tetrachloroaurate (III) with sodium borohydride,
wherein the sugar-immobilized metal nanoparticle is stable,
wherein a solution of the sugar-immobilized metal nanoparticle contains sodium tetrachloroaurate (III) at a concentration of 0.5 mM to 4 mM, sodium borohydride at a concentration that is 3 to 10 times a molar concentration of gold ion, and the ligand complex at a concentration of 10 mu M to 1000 mu M, and
wherein the sugar-immobilized metal nanoparticle forms a sugar-protein interactant through interaction of a sugar and a protein, and
wherein the sugar-immobilized metal nanoparticle forms a colloid solution when mixed with an aqueous solution.
[claim2]
2. A method for measuring sugar-protein interaction, comprising the step of forming a sugar-protein interactant through interaction of sugar and protein by mixing a solution containing the sugar-immobilized metal nanoparticle of claim 1 with a protein that recognizes the sugar situated at the end of the sugar-immobilized metal nanoparticle.
[claim3]
3. A method for recovering a protein from a sugar-protein interactant, comprising the steps of: forming a sugar-protein interactant through interaction of sugar and protein by mixing a solution containing the sugar-immobilized metal nanoparticle of claim 1 with a protein that recognizes the sugar situated at the end of the sugar-immobilized metal nanoparticle; and
adjusting a pH of an admixture solution of the sugar-protein interactant and water to 5 or less.
[claim4]
4. A method for recovering a protein from a sugar-protein interactant, comprising the steps of: forming a sugar-protein interactant through interaction of sugar and protein by mixing a solution containing the sugar-immobilized metal nanoparticle of claim 1 with a protein that recognizes the sugar situated at the end of the sugar-immobilized metal nanoparticle; and
mixing the sugar-protein interactant with a sugar that is recognizable by the protein.
  • Inventor, and Inventor/Applicant
  • SUDA YASUO
  • NISHIMURA TOMOAKI
  • KISHIMOTO YUKO
  • NAKAGAWA HIROMI
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
  • KAGOSHIMA UNIVERSITY
  • YASUO SUDA
IPC(International Patent Classification)
U.S. Cl./(Sub)
  • B82Y005/00
  • C07K001/22
  • C07K017/14
  • G01N033/543D6
  • G01N033/553
  • G01N033/58H4
  • S01N400/00
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