title: Expression and purification of neurotoxin-associated protein HA-33/A from Clostridium botulinum and evaluation of its antigenicity.
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.
authors.affiliation: Dept. of Biology, Faculty of Basic Science, Imam Hussein University, Tehran, Iran.
authors.forename: Ali
authors.initials: A
authors.lastname: Sayadmanesh
authors.affiliation: Dept. of Biology, Faculty of Basic Science, Imam Hussein University, Tehran, Iran.
authors.forename: Firouz
authors.initials: F
authors.lastname: Ebrahimi
authors.affiliation: Dept. of Biology, Faculty of Basic Science, Imam Hussein University, Tehran, Iran.
authors.forename: Abbas
authors.initials: A
authors.lastname: Hajizade
authors.affiliation: Dept. of Biology, Faculty of Basic Science, Imam Hussein University, Tehran, Iran.
authors.forename: Mosayeb
authors.initials: M
authors.lastname: Rostamian
authors.affiliation: Dept. of Biology, Faculty of Basic Science, Imam Hussein University, Tehran, Iran.
authors.forename: Hani
authors.initials: H
authors.lastname: Keshavarz
mesh_terms: D000818:Animals; D000942:Antigens, Bacterial / Q000276:immunology* / Q000302:isolation & purification* / Q000378:metabolism; D015153:Blotting, Western; D001905:Botulinum Toxins / Q000276:immunology* / Q000302:isolation & purification* / Q000378:metabolism; D003014:Clostridium botulinum / Q000737:chemistry* / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D051379:Mice; D009498:Neurotoxins / Q000276:immunology* / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D000942:Antigens, Bacterial; D009498:Neurotoxins; D011994:Recombinant Proteins; C118973:hemagglutinin 33 protein, Clostridium botulinum; D001905:Botulinum Toxins
keywords: Botulinum neurotoxin; Expression; Purification