Leshem Boaz, Sarfati Gadi, Novoa Andres, Breslav Igor, Marks Robert S
Department of Biotechnology Engineering, Ben-Gurion University of the Negev, PO Box 653, Beer Sheva 84105, Israel.
Luminescence. 2004 Mar-Apr;19(2):69-77. doi: 10.1002/bio.760.
We report herein a simple and effective way to photochemically immobilize biomolecules onto a fibre-optic silica surface. The system is based on a photoreactive benzophenone derivative that is bound to SiO2 surfaces of the optical fibre via a silane anchor. The benzophenone derivative was 4-allyloxybenzophenone, synthesized by standard procedures that were later used to synthesize the 4-(3'-chlorodimethylsilyl) propyloxybenzophenone and 4-(3'-dichloromethylsilyl) propyloxybenzophenone by regular hydrosilation procedures. After silanization with the benzophenone derivatives, the fibres were immersed in a cholera toxin B subunit solution and illuminated with UV light (wavelength > 345 nm). As a result of the photochemical reaction, a thin layer of the antigen was covalently bound to the benzophenone-modified surface. The photochemically modified fibre-optics were then tested as immunosensors in the detection of cholera anti-toxin antibody and revealed through chemiluminescence measurements. A secondary antibody labelled with horseradish peroxidase acted as the marker for the cholera toxin antibody. A photo-electronic set-up was designed specifically to monitor the signal. The immunosensor system was shown to be both specific and sensitive. The lowest rabbit serum titre detected was 1:1 700,000.
我们在此报告一种将生物分子光化学固定在光纤二氧化硅表面的简单有效方法。该系统基于一种光反应性二苯甲酮衍生物，它通过硅烷锚定与光纤的SiO2表面结合。二苯甲酮衍生物为4-烯丙氧基二苯甲酮，通过标准程序合成，随后通过常规硅氢化程序用于合成4-(3'-氯二甲基硅基)丙氧基二苯甲酮和4-(3'-二氯甲基硅基)丙氧基二苯甲酮。在用二苯甲酮衍生物进行硅烷化后，将光纤浸入霍乱毒素B亚基溶液中并用紫外光（波长>345nm）照射。光化学反应的结果是，一层薄薄的抗原共价结合到二苯甲酮修饰的表面上。然后将光化学修饰的光纤作为免疫传感器用于检测霍乱抗毒素抗体，并通过化学发光测量进行显示。用辣根过氧化物酶标记的二抗作为霍乱毒素抗体的标记物。专门设计了一个光电装置来监测信号。该免疫传感器系统显示出特异性和敏感性。检测到的最低兔血清滴度为1:1 700 000。
