Abstract:

Objective: Recombinant antibody expression in microorganism greatly facilitated the affinity and stability studies of recombinant antibodies, but previous studies had less reports about the selection of vectors. A suitable expression vector is important during the expression of the single chain antibody (scFv-H4) for aflatoxin B1. Methods: Four pET vectors such as pET20b, pET22b, pET28a and pET32a were employed in the functional expression of scFv-H4. Results: pET20b and pET22b vectors were able to express the functional antibody in periplasm. The vector of pET28a was not suitable for the expression of scFv-H4 due to lack of detectable scFv-H4 in total protein. Moreover, the detectable scFv-H4 was only expressed in inclusion body through pET32a expression system. Conclusion: pET22b was an excellent vector for the expression of scFv-H4, which might be a good reference for similar protein expression and be useful in the later study of scFv-H4 affinity and stability.

Key words: aflatoxin, single chain fragment, expression, Escherichia coli, pET