Pharmaceutical Development & Manufacturing Sciences, Janssen Research & Development, 2340 Beerse, Belgium.
Anal Chem. 2013 May 7;85(9):4770-6. doi: 10.1021/ac400514u. Epub 2013 Apr 16.
We report here on a new potentiometric biosensing principle for the detection of antibody-antigen interactions at the sensing membrane surface without the need to add a label or a reporter ion to the sample solution. This is accomplished by establishing a steady-state outward flux of a marker ion from the membrane into the contacting solution. The immunobinding event at the sensing surface retards the marker ion, which results in its accumulation at the membrane surface and hence in a potential response. The ion-selective membranes were surface-modified with an antibody against respiratory syncytial virus using click chemistry between biotin molecules functionalized with a triple bond and an azide group on the modified poly (vinyl chloride) group of the membrane. The bioassay sensor was then built up with streptavidin and subsequent biotinylated antibody. A quaternary ammonium ion served as the marker ion. The observed potential was found to be modulated by the presence of respiratory syncytial virus bound on the membrane surface. The sensing architecture was confirmed with quartz crystal microbalance studies, and stir effects confirmed the kinetic nature of the marker release from the membrane. The sensitivity of the model sensor was compared to that of a commercially available point-of-care test, with promising results.
我们在此报告一种新的电位生物传感原理，用于在无需向样品溶液中添加标记物或报告离子的情况下，在传感膜表面检测抗体-抗原相互作用。这是通过建立标记离子从膜内向接触溶液的稳态外向通量来实现的。传感表面上的免疫结合事件会阻碍标记离子，导致其在膜表面积累，从而产生电势响应。离子选择性膜通过点击化学将生物素分子与膜的聚（氯乙烯）基团上的叠氮基团连接，从而对其进行表面修饰，得到针对呼吸道合胞病毒的抗体。然后，使用链霉亲和素和随后的生物素化抗体构建生物测定传感器。季铵离子用作标记离子。观察到的电势受结合在膜表面上的呼吸道合胞病毒的存在而发生调制。通过石英晶体微天平研究证实了传感结构，搅拌效应证实了标记物从膜中释放的动力学性质。模型传感器的灵敏度与市售即时检测进行了比较，结果令人满意。
