化学学报 ›› 2008, Vol. 66 ›› Issue (4): 454-458. 上一篇    下一篇

研究论文

倏逝波光纤免疫传感器探头的修饰及表征

龙峰*, 施汉昌, 何苗, 朱安娜, 盛建武   

  1. (清华大学环境科学与工程系 环境模拟与污染控制国家重点联合实验室 北京 100084)
  • 投稿日期:2007-05-28 修回日期:2007-08-09 发布日期:2008-02-28
  • 通讯作者: 龙峰

Surface Modification and Characterization of the Probe Based on Evanescent Wave Fiber-optic Immunosensor

LONG Feng*; SHI Han-Chang; HE Miao; ZHU An-Na; SHENG Jian-Wu   

  1. (State Key Joint Laboratory of Environmental Simulation and Pollution Control, Department of Environmental Science and Engineering, Tsinghua University, Beijing 100084)
  • Received:2007-05-28 Revised:2007-08-09 Published:2008-02-28

光纤探头上修饰生物识别分子是倏逝波光纤免疫传感器实现目标物检测的关键步骤. 以微囊藻毒素-LR (microcystin-LR, MC-LR)为例, 采用先将小分子半抗原MC-LR与经戊二醛活化的惰性蛋白(OVA)反应生成复合物MC-LR-OVA, 然后将该复合物通过双功能试剂连接到硅烷化后的光纤探头表面, 并采用XPS和倏逝波全光纤免疫传感器对其修饰效果进行表征. 结果表明, 修饰后探头表面化学元素组成随不同修饰步骤发生了显著的变化, MC-LR-OVA被共价键合到探头表面上. 探头对MC-LR抗体表现出强烈的特异性响应, 而对其它抗体蛋白的非特异性吸附很弱, 并且具有良好的再生性能. 因此, 该修饰方案适用于环境小分子污染物的检测.

关键词: 光纤免疫传感器, 表面修饰, 探头, 倏逝波, 微囊藻毒素-LR

The modification of the surface of the fiber optic probe by bio-recognizing molecules is essential for detecting interest target by evanescent wave fiber optic immunosensor. In this study, microcystin-LR (MC-LR) was chosen as the example substance. Hapten-carrier conjugates MC-LR-Ovalbumin were synthesized by mixing MC-LR and ovalbumin (OVA) activated by glutaraldehyde, and then the conjugates were immobilized onto the silane layer on the probe with a heterobifunctional crosslinker. The probe modified was evaluated by XPS and an evanescent wave all-fiber immunosensor. The results showed that surface chemical compositions of the probe obviously changed at different modification steps, MC-LR-OVA was covalently bound with the probe surface, strong binding of antibody of MC-LR and low non-specific adsorption to other antibody was monitored, and good performance of regeneration was achieved. As a result, this modification method can be used for preparation of probe for the detection of low molecule pollutants.

Key words: fiber-optic immunosensor, surface modification, probe, evanescent wave, microcystin-LR