title: In-Vivo Neutralization of Botulinum Neurotoxin Serotype E Using Rabbit Polyclonal Antibody Developed against BoNT/E Light Chain.
abstract: BACKGROUND
  Clostridium botulinum is an obligate anaerobic, Gram positive bacterium that secretes extremely toxic substances known as botulinum neurotoxins (BoNTs) that cause serious paralytic illness called botulism. Based upon the serological properties, these neurotoxin have been classified into seven serotypes designated from A to G. Due to extreme toxicity of BoNTs, these neurotoxins have been designated as category A biowarfare agents. There is no commercial neutralizing antibody available for the treatment of botulism. Hence there is an urgent need to develop therapeutic intervention for prevention and cure of botulism within short period. BoNT antiserum injection is still the effective treatment.
  
  
  METHODS
  In the present study, the recombinant light chain of BoNT/E was successfully purified in soluble form. The purified rBoNT/E LC was used for the generation of polyclonal antibody in rabbit. In order to find out the neutralizing capacity of generated antisera, rabbit antiserum was incubated with 20 LD50 of botulinum neurotoxin type E for 1 hour at 37°C and then injected intraperitoneally (IP) into mice. Further in another set of experiments antiserum was administered in different ways that included administration of - antiserum and BoNT/E toxin simultaneously without preincubation, one after another at the same and different time points for its therapeutic ability. To find out cross neutralization capacity, rBoNT/E LC antiserum was pre-incubated with 5 LD50 of BoNT/A, BoNT/B, BoNT/F and then injected (IP) into mice. In all the cases mice were observed continuously for 96 hours.
  
  
  RESULTS
  The results clearly indicate that developed polyclonal rabbit antiserum showed serotype specific neutralization of BoNT/E toxin only but not of BoNT/A, BoNT/B and BoNT/F.
  
  
  CONCLUSIONS
  The developed antibodies will be used for preventive and therapeutic intervention of type 'E' botulism.
authors.affiliation: Biotechnology Division, Defence Research Development & Establishment, Jhansi Road, Gwalior- 474 002, M.P., India.
authors.forename: Sarita
authors.initials: S
authors.lastname: Rani
authors.affiliation: Biotechnology Division, Defence Research Development & Establishment, Jhansi Road, Gwalior- 474 002, M.P., India.
authors.forename: S
authors.initials: S
authors.lastname: Ponmariappan
authors.affiliation: Biotechnology Division, Defence Research Development & Establishment, Jhansi Road, Gwalior- 474 002, M.P., India.
authors.forename: Arti
authors.initials: A
authors.lastname: Sharma
authors.affiliation: Biotechnology Division, Defence Research Development & Establishment, Jhansi Road, Gwalior- 474 002, M.P., India.
authors.forename: D V
authors.initials: DV
authors.lastname: Kamboj
authors.affiliation: 2SOS in Zoology, Jiwaji University, Gwalior - 474 002, M.P., India.
authors.forename: A K
authors.initials: AK
authors.lastname: Jain
mesh_terms: D000818:Animals; D057134:Antibodies, Neutralizing / Q000276:immunology* / Q000627:therapeutic use; D001905:Botulinum Toxins / Q000037:antagonists & inhibitors / Q000276:immunology*; D001906:Botulism / Q000188:drug therapy / Q000276:immunology* / Q000382:microbiology; D003014:Clostridium botulinum / Q000276:immunology / Q000472:pathogenicity; D051379:Mice; D011817:Rabbits; D065288:Serogroup; D057127:Single-Chain Antibodies / Q000276:immunology
chemical_list: D057134:Antibodies, Neutralizing; D057127:Single-Chain Antibodies; D001905:Botulinum Toxins; C096326:botulinum toxin type E
keywords: Clostridium botulinum; biowarfare agent; botulinum neurotoxin; botulism; neutralizing antibody; therapeutic
  intervention