| Project/Area Number |
17380202
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Applied molecular and cellular biology
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| Research Institution | National Institute of Advanced Industrial Science and Technology (AIST) |
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Principal Investigator |
UZAWA Hirotaka National Institute of Advanced Industrial Science and Technology, Research Center of Advanced Bionics, Team Leader, バイオニクス研究センター, 研究チーム長 (60356566)
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| Co-Investigator(Kenkyū-buntansha) |
IZUMI Masayuki National Institute of Advanced Industrial Science and Technology, Research Center of Advanced Bionics, researcher, バイオニクス研究センター, 研究員 (80332641)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥15,200,000 (Direct Cost: ¥15,200,000)
Fiscal Year 2006: ¥5,800,000 (Direct Cost: ¥5,800,000)
Fiscal Year 2005: ¥9,400,000 (Direct Cost: ¥9,400,000)
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| Keywords | sugar / carbohydrate / Shiga toxins / chip / sensing / 生体機能利用 |
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| Research Abstract |
Cell-surface oligosaccharides linked to glycoproteins and glycosphingolipids play essential roles in a wide range of biological recognition events, such as cellular communication, signal transduction, protein folding and immune response. They also serve as toxin and viral receptors. In the present study, we apply the infection mechanism to the detection of toxins and/or viruses using synthetic sugar ligands. The significant results are summarized in below : (1) Synthesis of polynanionic glycopolymers carrying sugar units : several sugar units were synthesized such as Gb_2 for Shiga toxins, Man and Lac for E. coli, and also various sulfated sugars as model compounds. They have appropriate linkers with terminal amino groups at the reducing ends. Among them, the sugar units having Gb_2, Lac and Man were then reacted with poly(ethylene-alt-maleic anhydride) to give the corresponding polyanionic glycoplolymers. (2) Assembly of glyco-chips by alternating layer-by-layer adsorption : The glycopolymers having Gb_2, Lac and Man were immobilized onto chip substrates by the alternating layer-by-layer method. The chips were evidenced by FTIR-RAS method. (3) Shiga toxin detection applying the chips : SPR detection was examined using E. coli O157 Shiga toxins. Only the Gb_2 chips could bind the toxin strongly, while other chips did not. High content Gb_2 sugar polymer chip allowed to detect the toxin with high sensitivity. The detection limit was ca. 10ng/mL or less within 30 min. The present method is reliable and simple detection of the Shiga toxin, and can be applicable to the detection of other bacterial toxins or viruses. Further, during the present study, efficient synthetic methodology of monomeric sugars towards glyco chips have been developed with either a chemical or an enzymatic way.
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