title: Bioinformatics-based design of novel antigenic B-cell linear epitopes of Deinagkistrodon acutus venom.
abstract: OBJECTIVE
  With the help of bioinformatics analysis, we wished to develop a novel antivenom against the Deinagkistrodon (D.) acutus snake venom using B-cell linear epitopes of three primary toxins (serine protease, metalloprotease, and phospholipase A2).
  
  
  METHODS
  cDNA sequences of three toxins of D. acutus venom were retrieved from the NCBI database. B-cell linear epitopes were predicted using DNAStar and the website server software provided by IEDB. Then, the sequences of the predicted epitopes were artificially synthesized and inserted into the vector pET-32a-c(+). Recombinant antigen peptide was expressed and purified. BALB/c mice were immunized with the recombinant antigen peptide. The immunoprotective effect of this novel antivenom was measured by neutralization of venom haemorrhagic activity.
  
  
  RESULTS
  Six epitopes were obtained by bioinformatics analysis. ELISA analysis showed that antibody titre was >8,000 against snake venom and >64,000 against the recombinant peptide. Neutralization assays confirmed that the developed antivenom could effectively reduce the haemorrhagic activity of snake venom.
  
  
  CONCLUSIONS
  Six B-cell linear epitopes of D. acutus snake venom were predicted by bioinformatics analysis and successfully utilized to produce a novel antivenom.
authors.affiliation: Department of Emergency, Southwest Hospital, Third Military Medical University, Chongqing, China. minghua_liuIII@126.com.
authors.forename: Y-L
authors.initials: YL
authors.lastname: Cao
authors.forename: G-N
authors.initials: GN
authors.lastname: Guo
authors.forename: G-Y
authors.initials: GY
authors.lastname: Zhu
authors.forename: Z
authors.initials: Z
authors.lastname: Tian
authors.forename: Y-J
authors.initials: YJ
authors.lastname: Gou
authors.forename: C
authors.initials: C
authors.lastname: Chen
authors.forename: M-H
authors.initials: MH
authors.lastname: Liu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000997:Antivenins / Q000235:genetics* / Q000627:therapeutic use; D001483:Base Sequence; D019295:Computational Biology / Q000379:methods*; D015195:Drug Design; D018985:Epitopes, B-Lymphocyte / Q000235:genetics*; D006470:Hemorrhage / Q000139:chemically induced / Q000517:prevention & control; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D014688:Venoms / Q000235:genetics* / Q000633:toxicity
chemical_list: D000997:Antivenins; D018985:Epitopes, B-Lymphocyte; D014688:Venoms