Nanobody-based enzyme immunoassay for ochratoxin A in cereal with high resistance to matrix interference.

ABSTRACT: A sensitive indirect competitive nanobody-based enzyme linked immunosorbent assay (Nb-ELISA) for ochratoxin A (OTA) with high resistance to cereal matrix interference was developed. Nanobodies against OTA (Nb15, Nb28, Nb32, Nb36) were expressed in E. coli cells and their thermal stabilities were compared with that of an OTA-specific monoclonal antibody 6H8. All nanobodies could still retain their antigen-binding activity after exposure to temperature 95°C for 5min or to 90°C for 75min. Nb28 that exhibited the highest sensitivity in ELISA was selected for further research. An indirect competitive ELISA based on Nb28 was developed for OTA, with an IC₅₀ of 0.64ng/mL and a linear range (IC₂₀-IC₈₀) of 0.27-1.47ng/mL. Cereal samples were analyzed following a 2.5

SUBMITTER: Liu X 

PROVIDER: S-EPMC5862147 | biostudies-literature | 2017 Mar

REPOSITORIES: biostudies-literature