Characterisation of monoclonal antibody against aflatoxin B_1 produced in hybridoma 2C12 and its single-chain variable fragment expressed in recombinant Escherichia coli

摘要

An anti-aflatoxin B_1 monoclonal antibody (anti-AFB, mAb) from the hybridoma 2C12 was established and its inhibition concentration fifty (1C_(50)) for AFB, and relative cross-reactivities (CRs) to other mycotoxins were estimated to be 8 ng/mL and less than 4% compared with AFB, by a competitive direct enzyme-linked immunosorbent assay. For production of anti-AFB, single-chain variable fragment (anti-AFB_1, scFv) in recombinant Escherichia coli, its scFv-coding genes were cloned from the hybridoma 2C12. The anti-AFB, scFv formed inclusion bodies in the cytoplasm of E. coli required in vitro refolding process and hence recovered to retain binding activity successfully. Surface plasmon resonance analysis resulted that anti-AFB_1 scFv possessed 1.16 × 10~(-7) M of equilibrium dissociation constant (K_D), which was about 17 times higher than the parental anti-AFB, mAb of 6.95 × 10~(-9) M.