中国农业科学 ›› 2010, Vol. 43 ›› Issue (3): 565-570 .doi: 10.3864/j.issn.0578-1752.2010.03.017

• 贮藏·保鲜·加工 • 上一篇    下一篇

半抗原-载体蛋白偶联物的三种电泳鉴定方法

雷红涛,庞杰,贺丽苹,杨金易,孙远明,秦毅兰,梁明标

  

  1. (华南农业大学食品学院/广东省食品质量安全重点实验室)
  • 收稿日期:2008-11-27 修回日期:2009-11-05 出版日期:2010-02-10 发布日期:2010-02-10
  • 通讯作者: 孙远明

Identification of Hapten-Carrier Protein Conjugate by Three Electrophoresis Methods

LEI Hong-tao, PANG Jie, HE Li-ping, YANG Jin-yi, SUN Yuan-ming, QIN Yi-lan, LIANG Ming-biao
  

  1. (华南农业大学食品学院/广东省食品质量安全重点实验室)
  • Received:2008-11-27 Revised:2009-11-05 Online:2010-02-10 Published:2010-02-10
  • Contact: SUN Yuan-ming

摘要:

【目的】建立一种简单方便的半抗原-载体蛋白偶联物电泳鉴定方法。【方法】选用SDS-PAGE、非变性琼脂糖凝胶电泳、高效毛细管区带电泳3种方法对载体蛋白、偶联剂变性的载体蛋白、半抗原-载体蛋白进行比较鉴定,同时与光谱法结果比较。【结果】对分子量差异较小的载体蛋白和半抗原-载体蛋白偶联物,SDS-PAGE不能有效分辨;但偶联前后载体电荷有变化时,非变性琼脂糖凝胶电泳却能有效分辨游离的载体蛋白和半抗原-载体蛋白偶联物,相对SDS-PAGE表现出更高的分辨灵敏度;毛细管区带电泳能高效分辨载体和半抗原-载体蛋白偶联物,结果直观,但需较贵重的专用仪器。琼脂糖电泳和毛细管电泳法与可见-紫外光谱法鉴定结果一致。【结论】3种电泳方法各有优缺点,非变性凝胶电泳简便、廉价、有效,较适合普通实验室使用。

关键词: 半抗原-载体蛋白偶联物, SDS-PAGE, 非变性琼脂糖凝胶电泳, 毛细管区带电泳

Abstract:

【Objective】To establish a simple electrophoresis characterization method for hapten-carrier protein bioconjugate. 【Method】 SDS-PAGE, nondenaturing agarose gel electrophoresis and high performance capillary electrophoresis were employed and compared with spectrum method. 【Result】 The result showed that SDS-PAGE was not a sensitive candidate method to differentiate carrier protein from bioconjugate when both proteins have less molecular weight difference, but nondenaturing gel electrophoresis was a better alternative, which could discriminate the free carrier protein and bioconjugate protein with different net charge numbers. High performance capillary zone electrophoresis was another powerful approach for hapten-carrier protein bioconjugate characterization. The information obtained from three electrophoresis was in accordance with the one from ultraviolet- visible spectrum scanning. 【Conclusion】 Three electrophoresis methods had different advantages and disadvantages, cost-saving and simple nondenaturing electrophoresis seemed to have more availability in most laboratories.

Key words: hapten-carrier protein bioconjugate, SDS-PAGE, nondenaturing agarose gel electrophoresis, capillary zone electrophoresis