ABSTRACT
Botulinum neurotoxin [BoNT] complexes consist of neurotoxin and neurotoxin-associated proteins. Hemagglutinin-33 [HA-33] is a member of BoNT type A [BoNT/A] complex. Considering the protective role of HA-33 in preservation of BoNT/A in gastrointestinal harsh conditions and also its adjuvant role, recombinant production of this protein is favorable. Thus in this study, HA-33 was expressed and purified, and subsequently its antigenicity in mice was studied. Initially, ha-33 gene sequence of Clostridium botulinum serotype A was adopted from GenBank. The gene sequence was optimized and synthesized in pET28a [+] vector. E. coli BL21 [DE3] strain was transformed by the recombinant vector and the expression of HA-33 was optimized at 37°C and 5 h induction time. The recombinant protein was purified by nickel nitrilotriacetic acid agarose affinity chromatography and confirmed by immunoblotting. Enzyme Linked Immunoassay showed a high titer antibody production in mice. The results indicated a highly expressed and purified recombinant protein, which is able to evoke high antibody titers in mice