Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Tuwima 10, 10-748 Olsztyn, Poland.
Sensors (Basel). 2020 Apr 3;20(7):2014. doi: 10.3390/s20072014.
Suitable immobilization of a biorecognition element, such as an antigen or antibody, on a transducer surface is essential for development of sensitive and analytically reliable immunosensors. In this review, we report on (1) methods of antibody prefunctionalization using electroactive probes, (2) methods for immobilization of such conjugates on the surfaces of electrodes in electrochemical immunosensor construction and (3) the use of antibody-electroactive probe conjugates as bioreceptors and sensor signal generators. We focus on different strategies of antibody functionalization using the redox active probes ferrocene (Fc), anthraquinone (AQ), thionine (Thi), cobalt(III) bipyridine (Co(bpy)), Ru(bpy) and horseradish peroxidase (HRP). In addition, new possibilities for antibody functionalization based on bioconjugation techniques are presented. We discuss strategies of specific, quantitative antigen detection based on (i) a sandwich format and (ii) a direct signal generation scheme. Further, the integration of different nanomaterials in the construction of these immunosensors is presented. Lastly, we report the use of a redox probe strategy in multiplexed analyte detection.
生物识别元件（如抗原或抗体）在传感器表面的合适固定对于开发灵敏和分析可靠的免疫传感器至关重要。在这篇综述中，我们报告了（1）使用电活性探针对抗体进行预功能化的方法，（2）在电化学免疫传感器构建中固定此类缀合物在电极表面上的方法，以及（3）将抗体-电活性探针缀合物用作生物受体和传感器信号发生器的用途。我们专注于使用氧化还原活性探针[如二茂铁（Fc）、蒽醌（AQ）、硫堇（Thi）、钴（III）联吡啶（Co（bpy））、Ru（bpy）和辣根过氧化物酶（HRP）]对抗体进行功能化的不同策略。此外，还介绍了基于生物偶联技术的抗体功能化的新可能性。我们讨论了基于（i）三明治结构和（ii）直接信号产生方案的特异性、定量抗原检测策略。此外，还介绍了在这些免疫传感器构建中整合不同纳米材料的方法。最后，我们报告了在多重分析物检测中使用氧化还原探针策略的情况。
