Site-directed antibody immobilization techniques for immunosensors
Site-directed antibody immobilization techniques for immunosensors
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DOI：
10.1016/j.bios.2013.06.060
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发表时间：
2013-12-15
影响因子：
12.6
通讯作者：
Ramanaviciene, Almira
中科院分区：
文献类型：
--
作者：
Makaraviciute, Asta;Ramanaviciene, Almira
免疫传感器的灵敏度、再生性和稳定性直接取决于免疫传感器设计所使用的抗体类型、固定化分子的数量、固定化后的剩余活性以及传感界面上的正确方向。虽然以随机方式固定抗体制备的传感器表面获得了令人满意的结果，但传感分子的定点固定显著提高了免疫传感器的灵敏度，特别是当使用平面载体时。本文综述了免疫传感器设计中最常用的三种定点抗体固定化技术。将抗体固定在传感器表面的一种策略是通过与预先形成的Fc结合蛋白层的亲和作用，例如蛋白A、蛋白G、Fc区域特异性抗体或各种重组蛋白。另一种固定化策略是使用化学或基因工程抗体片段，这些抗体片段可以通过铰链区域中存在的巯基连接到覆盖着金色或自组装的单分子层的传感器表面。第三种最常见的策略是通过存在于抗体Fc区的氧化低聚糖部分来固定化抗体。综述了这些最常用的固定化技术的原理、优点、应用及存在的问题。(C)2013爱思唯尔B.V.保留所有权利。
Immunosensor sensitivity, regenerability, and stability directly depend on the type of antibodies used for the immunosensor design, quantity of immobilized molecules, remaining activity upon immobilization, and proper orientation on the sensing interface. Although sensor surfaces prepared with antibodies immobilized in a random manner yield satisfactory results, site-directed immobilization of the sensing molecules significantly improves the immunosensor sensitivity, especially when planar supports are employed. This review focuses on the three most conventional site-directed antibody immobilization techniques used in immunosensor design. One strategy of immobilizing antibodies on the sensor surface is via affinity interactions with a pre-formed layer of the Fc binding proteins, e.g., protein A, protein G, Fc region specific antibodies or various recombinant proteins. Another immobilization strategy is based on the use of chemically or genetically engineered antibody fragments that can be attached to the sensor surface covered in gold or self-assembled monolayer via the sulfhydryl groups present in the hinge region. The third most common strategy is antibody immobilization via an oxidized oligosaccharide moiety present in the Fc region of the antibody. The principles, advantages, applications, and arising problems of these most often applied immobilization techniques are reviewed. (C) 2013 Elsevier B.V. All rights reserved.