College of Food Science, South China Agricultural University, Guangzhou, China.
Methods Mol Biol. 2022;2446:547-554. doi: 10.1007/978-1-0716-2075-5_28.
Compared with traditional polyclonal and monoclonal antibodies, nanobodies derived from camelid heavy-chain antibodies have several advantages including small size, unique structure and binding geometry, high stability, and robust expression yields in numerous systems. Nanobody-based assays can also exhibit superior performance for immunodetection. Here, we describe protocols for three nanobody-based immunoassays for the detection of small chemical contaminants in environmental or agricultural samples: enzyme-linked immunosorbent assay (ELISA), fluorescence enzyme immunoassay (FEIA), and bioluminescent enzyme immunoassay (BLEIA). These methods are based on hapten-specific nanobodies, nanobody-alkaline phosphatase fusion proteins, and nanobody-nanoluciferase fusion proteins, respectively.
与传统的多克隆和单克隆抗体相比，骆驼重链抗体衍生的纳米抗体具有几个优势，包括体积小、独特的结构和结合几何形状、高稳定性以及在众多系统中产生稳健的表达产量。基于纳米抗体的检测也可以表现出优越的免疫检测性能。在这里，我们描述了三种基于纳米抗体的免疫检测方法，用于检测环境或农业样品中的小分子化学污染物：酶联免疫吸附测定（ELISA）、荧光酶免疫测定（FEIA）和生物发光酶免疫测定（BLEIA）。这些方法分别基于半抗原特异性纳米抗体、纳米抗体-碱性磷酸酶融合蛋白和纳米抗体-纳米荧光素酶融合蛋白。
