title: Comparative study of immunological and structural properties of two recombinant vaccine candidates against botulinum neurotoxin type E.
abstract: Recently, botulinum neurotoxin (BoNT)-derived recombinant proteins have been suggested as potential botulism vaccines. Here, with concentrating on BoNT type E (BoNT/E), we studied two of these binding domain-based recombinant proteins: a multivalent chimer protein, which is composed of BoNT serotypes A, B and E binding subdomains, and a monovalent recombinant protein, which contains 93 amino acid residues from recombinant C-terminal heavy chain of BoNT/E (rBoNT/E-HCC). Both proteins have an identical region (48 aa) that contains one of the most important BoNT/E epitopes (YLTHMRD sequence).
  
  
  
  The recombinant protein efficiency in antibody production, their structural differences, and their BoNT/E-epitope location were compared by using ELISA, circular dichroism, computational modeling, and hydrophobicity predictions.
  
  
  
  Immunological studies indicated that the antibody yield against rBoNT/E-HCC was higher than chimer protein. Cross ELISA confirmed that the antibodies against the chimer protein recognized rBoNT/E-HCC more efficiently. However, both antibody groups (anti-chimer and anti-rBoNT/E-HCC antibodies) were able to recognize other proteins. Structural studies with circular dichroism showed that chimer proteins have slightly more secondary structures than rBoNT/E-HCC.
  
  
  
  The immunological results suggested that the above-mentioned identical region in rBoNT/E-HCC is more exposed. Circular dichroism, computational protein modeling and hydrophobicity predictions indicated a more exposed location for the identical region in rBoNT/E-HCC than the chimer protein, which is strongly in agreement with immunological results.
authors.affiliation: Dept. of Biology, Faculty of Basic Sciences, Imam Hussein University, Tehran, Iran. jmosavi@ihu.ac.ir
authors.forename: Mosayeb
authors.initials: M
authors.lastname: Rostamian
authors.affiliation: Dept. of Biology, Faculty of Basic Sciences, Imam Hussein University, Tehran, Iran. jmosavi@ihu.ac.ir
authors.forename: Seyed Jafar
authors.initials: SJ
authors.lastname: Mousavy
authors.affiliation: Dept. of Biology, Faculty of Basic Sciences, Imam Hussein University, Tehran, Iran. jmosavi@ihu.ac.ir
authors.forename: Firouz
authors.initials: F
authors.lastname: Ebrahimi
authors.affiliation: Dept. of Biology, Faculty of Sciences, Razi University, Kermanshah, Iran.|Medical Biology Research Center, Kermanshah University of Medical Science, Kermanshah, Iran.
authors.forename: Seyyed Abolghasem
authors.initials: SA
authors.lastname: Ghadami
authors.affiliation: Dept. of Ophthalmology and Visual Sciences and Pharmacology, University of Wisconsin School of Medicine and Public Health, Madison, WI, USA.
authors.forename: Nader
authors.initials: N
authors.lastname: Sheibani
authors.affiliation: Dept. of Biology, Faculty of Basic Sciences, Imam Hussein University, Tehran, Iran. jmosavi@ihu.ac.ir
authors.forename: Mohammad Ebrahim
authors.initials: ME
authors.lastname: Minaei
authors.affiliation: Dept. of Biology, Faculty of Basic Sciences, Imam Hussein University, Tehran, Iran. jmosavi@ihu.ac.ir
authors.forename: Mohammad Ali
authors.initials: MA
authors.lastname: Arefpour Torabi
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D001428:Bacterial Vaccines / Q000276:immunology*; D001905:Botulinum Toxins / Q000737:chemistry / Q000276:immunology*; D002942:Circular Dichroism; D004797:Enzyme-Linked Immunosorbent Assay; D007074:Immunoglobulin G / Q000097:blood; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000276:immunology*; D014614:Vaccines, Synthetic / Q000276:immunology
chemical_list: D000907:Antibodies, Bacterial; D001428:Bacterial Vaccines; D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; D014614:Vaccines, Synthetic; D001905:Botulinum Toxins; C096326:botulinum toxin type E