One-Step Ultrasensitive Bioluminescent Enzyme Immunoassay Based on Nanobody/Nanoluciferase Fusion for Detection of Aflatoxin B₁ in Cereal.

ABSTRACT: Nanoluciferase (Nluc), the smallest luciferase known, was used as the fusion partner with a nanobody against aflatoxin B₁ to develop a bioluminescent enzyme immunoassay (BLEIA) for detection of the aflatoxin B₁ in cereal. Nanobody (clone G8) against aflatoxin B₁ was fused with nanoluciferase and cloned into a pET22b expression vector, and then transformed into Escherichia coli. The nanobody fusion gene contained a hexahistidine tag for purification by immobilized metal affinity chromatography, yielding a biologically active fusion protein. The fusion protein G8-Nluc retained binding properties of the original nanobody. Concentration of the coelenterazine substrate and buffer composition were also optimized to provide high intensity and long half-life of the

SUBMITTER: Ren W 

PROVIDER: S-EPMC7792509 | biostudies-literature | 2019 May

REPOSITORIES: biostudies-literature