高等学校化学学报

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表面等离子体共振成像法用于糖蛋白分析

刘巍1,2, 陈义1   

    1. 中国科学院化学研究所生命分析化学实验室, 北京 100190;
    2. 中国科学院研究生院, 北京 100049
  • 收稿日期:2008-02-28 修回日期:1900-01-01 出版日期:2008-09-10 发布日期:2008-09-10
  • 通讯作者: 陈义

Glycoproteins Analysis by Surface Plasmon Resonance Absorption and Imaging

LIU Wei1,2, CHEN Yi1*   

    1. Laboratory of Analytical Chemistry for Life Science, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, China;
    2. Graduate School, Chinese Academy of Sciences, Beijing 100049, China
  • Received:2008-02-28 Revised:1900-01-01 Online:2008-09-10 Published:2008-09-10
  • Contact: CHEN Yi

摘要: 建立了一套表面等离子体共振成像方法, 用于筛选糖蛋白和非糖蛋白, 并可以区分不同糖蛋白与伴刀豆凝集素的识别强度. 波长调制的表面等离子体共振与成像法所得结果一致. 伴刀豆凝集素的洗脱成像实验为识别强度提供了补充验证方案.

关键词: 糖蛋白, 蛋白质微阵列, 表面等离子体共振成像

Abstract: Surface plasmon resonance(SPR) absorption and imaging(SPRI) have been explored for screening and differentiating glycoproteins from non-glycoprotein according to the shift of maximum SPR absorption and/or to the variation of picture gray intensity. In combination with competitive washing with sugar solutions, SPRI has the ability to produce different signals for different gylcoproteins, offering a potential way to identify these gylcoconjugates. To conduct SPR and SPRI, proteins(at 1.0 mg/mL), in stead of lectins, were immobilized on gold sensing films to reduce the use of the costly glycoproteins. The sensing film was fixed in a laboratory-established SPR or SPRI system, equilibrated with 50 mmol/L Tris-HCl at pH=7.4 at a flow rate of 20 μL/min, reacted with 200 nmol/L ConA(prepared in the Tris-HCl solution containing 120 mmol/L NaCl, 25 mmol/L CaCl2 and 5 mmol/L MnCl2) at room temperature for 1 h and rewashed with Tris-HCl solution. For competitive washing, 80 mmol/L glucose solution was pumped into the analytical cell and reacted for 1 h. The signals were recorded whenever necessary. Non-gylcoproteins had no signals when using either SPR or SPRI while glycoproteins gave different SPR shifts and gray intensity in SPRI.

Key words: Glycoprotein, Protein microarray, Surface plasmon resonance imaging(SPRI)

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