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Practical Application of Fluorescence Labeling to Component Sugar Analysis of Glycoproteins

Practical Application of Fluorescence Labeling to Component Sugar Analysis of Glycoproteins
荧光标记在糖蛋白成分糖分析中的实际应用
批准号:
62880019
负责人:
HASE Sumihiro
金额:
$8.77万
依托单位:
依托单位国家:
日本
项目类别:
Grant-in-Aid for Developmental Scientific Research
财政年份:
1987
资助国家:
日本
项目状态:
已结题
起止时间:
1987 至 1988

项目摘要

项目成果

HASE Sumihiro的其他基金

相关文献

中文摘要
翻译
报道了一种用吡啶层析法测定糖蛋白组成糖的方法(H.Takemoto等人)。1985年)。该方法包括糖蛋白的酸解、氨基的N-乙酰化、单糖还原端用NaBH_3CN、2-氨基吡啶和HCl的吡啶层析、用凝胶色谱(HPLC)纯化糖的吡啶氨基(PA-)衍生物,以及用反相高效液相色谱法分析PA-糖。为研制全自动糖分析仪,对吡啶层析条件进行了改进。1)二甲胺等挥发性试剂。在NaBH_3CN和2-氨基吡啶-盐酸盐之间使用了硼烷络合物和2-氨基吡啶-冰醋酸。2)对反应条件进行了选择,以获得最高的PA-糖得率(中性糖70%,N-乙酰氨基糖87%)。3)为了有效地蒸发试剂,开发了减压氮气流动。采用新的反应条件和蒸发过程,可以省去凝胶层析(HPLC)纯化PA-糖的步骤。在此基础上,建立了一套新条件下的自动吡啶层析装置,并用配有自动进样器和数据处理机的高效液相色谱仪对得到的PA-糖进行了分析。糖蛋白酸性水解物中糖的用量为1-10nmol时,结果较为合理。
英文摘要
A method for determination of component sugars of glycoproteins by pyridylamination was reported (H. Takemoto, et al. 1985). The method includes acid hydrolysis of glycoprotein, N-acetylation of amino groups, pyridylamination of the reducing ends of the monosaccharides with NaBH_3CN, 2-aminopyridine, and HCl, purification of pyridylamino (PA-) derivatives of sugars by gel chromatography (HPLC), and the analysis of PA-sugars by reversed-phase HPLC. The pyridylamination conditions were improved for the purpose of the making of an automatic sugar analyzer. 1) Volatile reagents such as dimethylamine. borane complex, and 2-aminopyridine-acetic acid were used intead of NaBH_3CN and 2-aminopyridine-HCl. 2) The reaction conditions were chosen to obtain maximum yield of PA-sugars (neutral sugars 70% and N-acetylaminosugars 87%). 3) Nitrogen flow under reduced pressure was developed to evaporate the reagents effectively. Using the new reaction conditions and the evaporation procedre, purification of PA-sugars by gel chromatography (HPLC) could be omitted. An automatic pyridylamination apparatus using the new conditions was constructed and PA-sugars thus obtained were analyzed by HPLC equipped with an automatic injector and a data processor. Reasonable results were obtained by using 1-10 nmol of sugars of acid hydrolysates of glycoproteins.
期刊论文(43)
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会议论文
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通讯作者:
S.Hase: Anal.Biochem.67. 321-326 (1987)
S.Hase:Anal.Biochem.67。
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通讯作者:
S. Hase: "A New Trisaccharide Sugar Chain Linked to a Serine Residue in Bovine Blood Coagulation Factors VII and IX" J. Biochem.104. 867-868 (1988)
S. Hase:“与牛凝血因子 VII 和 IX 中的丝氨酸残基相连的新三糖糖链”J. Biochem.104。
DOI: --
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通讯作者:
S.Hase: J.Biochem.104. 867-868 (1988)
S.Hase:J.Biochem.104。
DOI: --
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共 16 条
    Analysis of Oligosaccharide Structures from the Reducing End Terminal by Combining Partial Acid Hydrolysis and a Two-Dimensional Sugar Map
    • 批准号:
      10558100
    • 项目类别:
      Grant-in-Aid for Scientific Research (B)
    • 资助金额:
      $5.06万
    • 财政年份:
      1998
    • 负责人:
      HASE Sumihiro
    • 依托单位:
    Analysis of activation of neutral alpha-mannosidase activated with Co(II)
    • 批准号:
      05453211
    • 项目类别:
      Grant-in-Aid for General Scientific Research (B)
    • 资助金额:
      $4.74万
    • 财政年份:
      1993
    • 负责人:
      HASE Sumihiro
    • 依托单位:
    Search for xylosylglucose-type sugar chains by pyridylamination
    • 批准号:
      03453164
    • 项目类别:
      Grant-in-Aid for General Scientific Research (B)
    • 资助金额:
      $4.42万
    • 财政年份:
      1991
    • 负责人:
      HASE Sumihiro
    • 依托单位:
    Sensitive analysis of O-linked sugar chains by fluorescence labeling
    • 批准号:
      01470147
    • 项目类别:
      Grant-in-Aid for General Scientific Research (B)
    • 资助金额:
      $4.35万
    • 财政年份:
      1989
    • 负责人:
      HASE Sumihiro
    • 依托单位: