Nanobody-Based Assays for the Detection of Environmental and Agricultural Contaminants

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.