In-Vivo Neutralization of Botulinum Neurotoxin Serotype E Using Rabbit Polyclonal Antibody Developed against BoNT/E Light Chain. 2017

Sarita Rani, and S Ponmariappan, and Arti Sharma, and D V Kamboj, and A K Jain
Biotechnology Division, Defence Research Development & Establishment, Jhansi Road, Gwalior- 474 002, M.P., India.

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.

UI MeSH Term Description Entries
D011817 Rabbits A burrowing plant-eating mammal with hind limbs that are longer than its fore limbs. It belongs to the family Leporidae of the order Lagomorpha, and in contrast to hares, possesses 22 instead of 24 pairs of chromosomes. Belgian Hare,New Zealand Rabbit,New Zealand Rabbits,New Zealand White Rabbit,Rabbit,Rabbit, Domestic,Chinchilla Rabbits,NZW Rabbits,New Zealand White Rabbits,Oryctolagus cuniculus,Chinchilla Rabbit,Domestic Rabbit,Domestic Rabbits,Hare, Belgian,NZW Rabbit,Rabbit, Chinchilla,Rabbit, NZW,Rabbit, New Zealand,Rabbits, Chinchilla,Rabbits, Domestic,Rabbits, NZW,Rabbits, New Zealand,Zealand Rabbit, New,Zealand Rabbits, New,cuniculus, Oryctolagus
D001905 Botulinum Toxins Toxic proteins produced from the species CLOSTRIDIUM BOTULINUM. The toxins are synthesized as a single peptide chain which is processed into a mature protein consisting of a heavy chain and light chain joined via a disulfide bond. The botulinum toxin light chain is a zinc-dependent protease which is released from the heavy chain upon ENDOCYTOSIS into PRESYNAPTIC NERVE ENDINGS. Once inside the cell the botulinum toxin light chain cleaves specific SNARE proteins which are essential for secretion of ACETYLCHOLINE by SYNAPTIC VESICLES. This inhibition of acetylcholine release results in muscular PARALYSIS. Botulin,Botulinum Neurotoxin,Botulinum Neurotoxins,Clostridium botulinum Toxins,Botulinum Toxin,Neurotoxin, Botulinum,Neurotoxins, Botulinum,Toxin, Botulinum,Toxins, Botulinum,Toxins, Clostridium botulinum
D001906 Botulism A disease caused by potent protein NEUROTOXINS produced by CLOSTRIDIUM BOTULINUM which interfere with the presynaptic release of ACETYLCHOLINE at the NEUROMUSCULAR JUNCTION. Clinical features include abdominal pain, vomiting, acute PARALYSIS (including respiratory paralysis), blurred vision, and DIPLOPIA. Botulism may be classified into several subtypes (e.g., food-borne, infant, wound, and others). (From Adams et al., Principles of Neurology, 6th ed, p1208) Botulism, Infantile,Botulism, Toxico-Infectious,Clostridium botulinum Infection,Foodborne Botulism,Infant Botulism,Toxico-Infectious Botulism,Wound Botulism,Botulism, Foodborne,Botulism, Infant,Botulism, Toxico Infectious,Botulism, Wound,Clostridium botulinum Infections,Foodborne Botulisms,Infant Botulisms,Infantile Botulism,Infection, Clostridium botulinum,Toxico Infectious Botulism,Wound Botulisms
D003014 Clostridium botulinum A species of anaerobic, gram-positive, rod-shaped bacteria in the family Clostridiaceae that produces proteins with characteristic neurotoxicity. It is the etiologic agent of BOTULISM in humans, wild fowl, HORSES; and CATTLE. Seven subtypes (sometimes called antigenic types, or strains) exist, each producing a different botulinum toxin (BOTULINUM TOXINS). The organism and its spores are widely distributed in nature.
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D051379 Mice The common name for the genus Mus. Mice, House,Mus,Mus musculus,Mice, Laboratory,Mouse,Mouse, House,Mouse, Laboratory,Mouse, Swiss,Mus domesticus,Mus musculus domesticus,Swiss Mice,House Mice,House Mouse,Laboratory Mice,Laboratory Mouse,Mice, Swiss,Swiss Mouse,domesticus, Mus musculus
D057127 Single-Chain Antibodies A form of antibodies consisting only of the variable regions of the heavy and light chains (FV FRAGMENTS), connected by a small linker peptide. They are less immunogenic than complete immunoglobulin and thus have potential therapeutic use. Fv Antibody Fragments, Single-Chain,ScFv Antibodies,Single-Chain Fv,Single-Chain Fv Antibody,Single-Chain Fv Antibody Fragments,Single-Chain Variable Fragment,Single-Chain Variable Fragment Antibodies,Single-Chain Variable Fragment Antibody,Single-Chain Variable Fragments,Antibodies, ScFv,Antibodies, Single-Chain,Antibody, Single-Chain Fv,Fragment, Single-Chain Variable,Fragments, Single-Chain Variable,Fv Antibody Fragments, Single Chain,Fv Antibody, Single-Chain,Fv, Single-Chain,Single Chain Antibodies,Single Chain Fv,Single Chain Fv Antibody,Single Chain Fv Antibody Fragments,Single Chain Variable Fragment,Single Chain Variable Fragment Antibodies,Single Chain Variable Fragment Antibody,Single Chain Variable Fragments,Variable Fragment, Single-Chain,Variable Fragments, Single-Chain
D057134 Antibodies, Neutralizing Antibodies that reduce or abolish some biological activity of a soluble antigen or infectious agent, usually a virus. Neutralizing Antibodies,Antibody, Neutralizing,Neutralizing Antibody
D065288 Serogroup A set of variants within a species of microorganisms that are antigenically, closely related. With bacteria, a serogroup refers to a group that shares a common antigen. Serotype,Serovar,Serogroups,Serotypes,Serovars

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