Agkistrodon acutus bites are conventionally treated with animal-derived antivenom, the use of which is limited due to allergic reactions and serum sickness. Thus in the present study, the genes of humanized antibodies produced in response to A. acutus venom were extracted from lymphocytes from patients bitten by A. acutus. A single-chain variable fragment (scFv) library against venom was constructed using a T7 phage display system. ScFv genes that exhibited high affinity to venom were selected by library biopanning. An expression system was constructed for antivenom scFv fused with 6×His tag at its N- and C-terminus using pET-28a (+) vector. The scFv proteins could achieve functional and soluble expression in Escherichia coli via the auto-induction method. The purity and activity of the scFv genes and proteins were confirmed by SDS-PAGE, western blotting and ELISA. The results indicated that three soluble scFv proteins exhibited specific affinity to A. acutus venom and were harvested via the auto-induction method.
Keywords: Agkistrodon acutus venom; auto-induction; phage display; single-chain variable fragment.