| 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 |
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| 引用本文： | 王清刚,罗国安,杨树标. 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[J]. 高等学校化学学报, 1999, 20(10): 1551-1553 |
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| 作者姓名： | 王清刚 罗国安 杨树标 |
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| 作者单位： | 1. 清华大学化学系, 北京 100084;2. 香港大学妇产科系, 香港 |
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| 摘 要： | 生物分子之间的特异性相互作用是生物界普遍存在的现象．研究这些现象，对揭示生物化学作用机理、药物研究等具有重要意义．结合常数Ｋｂ是描述生物分子之间特异性相互作用最主要的参数，测定结合常数的传统方法包括平衡透析、凝胶过滤色谱和分光光度法等［１］．亲和毛细管电泳（Ａｆｆｉｎｉｔｙｃａｐｉｌｌａｒｙｅｌｅｃｔｒｏｐｈｏｒｅｓｉｓ，简称ＡＣＥ）是近几年发展起来的毛细管电泳的一个分支，在研究生物分子之间特异性相互作用等方面有很好的应用前景［２～５］．与上述传统方法相比，ＡＣＥ具有测定速度快，样品用量少，有多…
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| 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 |
| 收稿时间： | 1999-02-08 |
| 修稿时间： | 1999-02-08 |
Determination of the Binding Constant Between Bovine Serum Albumin and Its Monoclonal Antibody by Affinity Capillary Electrophoresis with Laser Induced Fluorescence Detection |
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| WANG Qing-gang,LUO Guo-An,William S.B. Yeung. Determination of the Binding Constant Between Bovine Serum Albumin and Its Monoclonal Antibody by Affinity Capillary Electrophoresis with Laser Induced Fluorescence Detection[J]. Chemical Research In Chinese Universities, 1999, 20(10): 1551-1553 |
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| Authors: | WANG Qing-gang LUO Guo-An William S.B. Yeung |
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| Affiliation: | 1. Department of Chemistry, Tsinghua University, Beijing 100084, China;2. Department of Obstetrics and Gynecology, the University of Hong Kong, Hong Kong, China |
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| 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. |
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| Keywords: | Affinity capillary electrophoresis Laser induced fluorescence detection Protein Binding constant |
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