高等学校化学学报 ›› 1999, Vol. 20 ›› Issue (10): 1551.

• 研究简报 • 上一篇    下一篇

亲和毛细管电泳激光诱导荧光法测定牛血清白蛋白与其单克隆抗体的结合常数

王清刚1, 罗国安1, 杨树标2   

  1. 1. 清华大学化学系, 北京 100084;
    2. 香港大学妇产科系, 香港
  • 收稿日期:1999-02-08 出版日期:1999-10-24 发布日期:1999-10-24
  • 通讯作者: 罗国安
  • 作者简介:王清刚,男,26岁,博士研究生.
  • 基金资助:

    国家自然科学基金(批准号:692350220)重点项目资助课题

Determination of the Binding Constant Between Bovine Serum Albumin and Its Monoclonal Antibody by Affinity Capillary Electrophoresis with Laser Induced Fluorescence Detection

WANG Qing-Gang1, LUO Guo-An1, William S. B. Yeung2   

  1. 1. Department of Chemistry, Tsinghua University, Beijing 100084, China;
    2. Department of Obstetrics and Gynecology, the University of Hong Kong, Hong Kong, China
  • Received:1999-02-08 Online:1999-10-24 Published:1999-10-24

关键词: 亲和毛细管电泳, 激光诱导荧光检测, 蛋白质, 结合常数

Abstract: The binding constant(Kb) between fluorescein isothiocyanate labeled bovine serum albumin(FITC-BSA) and its monoclonal antibody was determined using the ligand separation mode of affinity capillary electrophoresis(ACE) with laser induced fluorescence detection(LIF). Free FITC-BSA and the complexes were separated by capillary electrophoresis in 7min after incubation. The running buffer was 100 mmol/L HEPE Sat pH7.4, and separation voltage was 30kV. The Kbwas determined to be 4.7×108L/mol using Scatchard plotting method. The influences of incubation time and electrophoresis conditions on determination were also studied.

Key words: Affinity capillary electrophoresis, Laser induced fluorescence detection, Protein, Binding constant

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