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BIOMOLECULE-IMMOBILIZED CARRIER AND METHOD FOR IMMOBILIZING BIOMOLECULE ON CARRIER

Foreign code F110002785
File No. S2008-0336-C0
Posted date Apr 15, 2011
Country WIPO
International application number 2009JP053038
International publication number WO 2009104738
Date of international filing Feb 20, 2009
Date of international publication Aug 27, 2009
Priority data
  • P2008-039890 (Feb 21, 2008) JP
Title BIOMOLECULE-IMMOBILIZED CARRIER AND METHOD FOR IMMOBILIZING BIOMOLECULE ON CARRIER
Abstract Provided are a biomolecule-immobilized carrier obtained by immobilizing a biomolecule having a membrane-binding peptide containing a C terminal polypeptide of fucosyltransferase derived from Helicobacter pylori or a membrane-binding molecule having the membrane-binding peptide and various biomolecules and a method for immobilizing a biomolecule to a carrier, particularly a biomolecule-immobilized magnetic microparticle, a probe-immobilized carrier using the biomolecule and/or the membrane-binding molecule are/is used as a probe, and a separating agent for a biomolecule to which the biomolecule-immobilized magnetic microparticle is applied. A biomolecule having a membrane-binding peptide containing either an amino acid sequence represented by SEQ ID NO: 1 or an amino acid sequence encoded by a base sequence represented by SEQ ID NO: 2 and/or a membrane-binding molecule having the membrane-binding peptide and the biomolecule are/is attached to a carrier coated with an organic membrane containing a phospholipid.
Outline of related art and contending technology BACKGROUND ART
The magnetic nanoparticle can be recovered by a magnetic, conventionally, the magnetic fine particles to which the antibody and the like as an enzyme or biological molecule is immobilized and an attempt is made. For example, Japanese Patent Application Laid-Open 10-75780 and Japanese Patent Application Laid-Open 11-243951 is, by the chemical method were fixed directly in the magnetic fine particles disclosed (Patent Document 1 and 2). In addition, in Japanese Patent Application Laid-Open 2005-289822, the peptide is anchored to a membrane derived from a fuzed magnetic bacteria (magnetosomes) the magnetic fine particles is disclosed (Patent Document 3) and, in the Japanese Patent Application Laid-Open 2006-75103, and the IgG-binding peptide Mms13 anchored proteins in vivo that a fusion protein expressed in a membrane derived from a fuzed magnetosomes magnetic bacteria is disclosed (Patent Document 4).
On the other hand, some conventional cell adhesion polypeptides are known, for example, an extracellular sugar chains binding laminin derived polypeptide thin decane (Non-Patent Document 1), an integrin ligand Arg-Gly-Asp sequence (RDG) fibronectin (Non-Patent Document 2) tripeptide sequence, or, as a polypeptide having a cell membrane adhesion, Helicobacter, α 1, 3 - and α 1, 4 - fucose GnTDI pylori derived from a C-terminal of the fucose transferase polypeptide (non-patent documents 3 and 4) can be exemplified. The Helicobacter, α 1 derived from H. pylori, 3 / α 1, the C-terminal polypeptides GnTDI fucose 4 -, α over helix structure which can be adhered to the cell membrane is known (Non-Patent Document 3).
Scope of claims (In Japanese)請求の範囲 [1]
 配列番号1に示すアミノ酸配列および配列番号2に示す塩基配列がコードするアミノ酸配列のいずれかを含む膜接着ペプチドを有する生体分子、および/または前記膜接着ペプチドと生体分子とを有する膜接着分子を、リン脂質を含む有機膜で被覆された担体に接着させてなる、生体分子固定化担体。

[2]
 前記担体が磁性微粒子、もしくは前記膜接着ペプチドを有する生体分子および/または前記膜接着分子をプローブとするプローブ固定化担体である、請求項1に記載の生体分子固定化担体。

[3]
 前記生体分子が酵素または糖鎖である、請求項1または請求項2に記載の生体分子固定化担体。

[4]
 前記酵素が糖転移酵素または糖加水分解酵素である、請求項3に記載の生体分子固定化担体。

[5]
 配列番号1に示すアミノ酸配列および配列番号2に示す塩基配列がコードするアミノ酸配列のいずれかを含む膜接着ペプチドを有する生体分子、および/または前記膜接着ペプチドと生体分子とを有する膜接着分子を、リン脂質を含む有機膜で被覆された磁性微粒子に接着させてなる、生体分子の分離剤。

[6]
 担体に生体分子の生理活性を保持した状態で生体分子を直接または間接に固定化させる方法であって、
 担体に固定するための固定リガンドをリン脂質に修飾するリン脂質修飾ステップと、
 前記固定リガンドを修飾したリン脂質を用いて担体表面に有機膜を形成して被覆させる担体被覆ステップと、
 配列番号1に示すアミノ酸配列および配列番号2に示す塩基配列がコードするアミノ酸配列のいずれかを含む膜接着ペプチドを有する生体分子、および/または前記膜接着ペプチドをリンカーとして融合ないし結合した生体分子を前記有機膜に接着させる接着ステップと
 を有する生体分子担体固定化方法。

  • Applicant
  • ※All designated countries except for US in the data before July 2012
  • National University Corporation Hokkaido University
  • Inventor
  • NISHIMURA, Shin-Ichiro
  • NARUCHI, Kentaro
IPC(International Patent Classification)
Specified countries National States: AE AG AL AM AO AT AU AZ BA BB BG BH BR BW BY BZ CA CH CN CO CR CU CZ DE DK DM DO DZ EC EE EG ES FI GB GD GE GH GM GT HN HR HU ID IL IN IS JP KE KG KM KN KP KR KZ LA LC LK LR LS LT LU LY MA MD ME MG MK MN MW MX MY MZ NA NG NI NO NZ OM PG PH PL PT RO RS RU SC SD SE SG SK SL SM ST SV SY TJ TM TN TR TT TZ UA UG US UZ VC VN ZA ZM ZW
ARIPO: BW GH GM KE LS MW MZ NA SD SL SZ TZ UG ZM ZW
EAPO: AM AZ BY KG KZ MD RU TJ TM
EPO: AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LT LU LV MC MK MT NL NO PL PT RO SE SI SK TR
OAPI: BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG
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