Isolation and characterization of a neutralizing antibody specific to internalization domain of Clostridium botulinum neurotoxin type B.
Botulinum neurotoxins (BoNTs), the causative agents for life-threatening human disease botulism, have been recognized as biological warfare agents. In this study, a neutralizing mouse monoclonal antibody against botulinum neurotoxin serotype B (BoNT/B), named BTBH-N1, was developed from mice immunized with BoNT/B toxoid without non-toxic components, which are generally associated with the toxin. Western blot analysis, using recombinant toxin fragments containing light (L), N-terminal half of heavy (HN) and C-terminal half of heavy chains, indicated that BTBH-N1 recognizes linear epitopes located on the HN domain. An in vivo neutralization assay with mice, was conducted to characterize the neutralization capacity of the BTBH-N1. Only 10 microg of BTBH-N1 completely neutralized 20 units (1 unit = one 50% lethal dose) of BoNT/B. Even though the Mab (up to 100 microg) failed to protect mice challenged with 100 units, it significantly prolonged the time to death in a dose dependent manner. BTBH-N1, the first neutralizing antibody against BoNT/B, could be further developed as effective biological therapeutics for preventing and treating botulism, as well as other diseases caused by BoNT/B.
Yang
Gi-Hyeok
GH
Microbial Toxin Research Institute, Chungnam 336-708, South Korea.
Kim
Kyu-Sik
KS
Kim
Hak-Woo
HW
Jeong
Sung Tae
ST
Huh
Gyung Heng
GH
Kim
Ji Cheon
JC
Jung
Hyun Ho
HH
D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification* / Q000378:metabolism; D015153:Blotting, Western; D001905:Botulinum Toxins / Q000378:metabolism*; D019274:Botulinum Toxins, Type A; D001906:Botulism / Q000276:immunology / Q000517:prevention & control*; D003014:Clostridium botulinum / Q000737:chemistry*; D003429:Cross Reactions / Q000276:immunology; D004195:Disease Models, Animal; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000378:metabolism; D007114:Immunization; D051379:Mice; D009500:Neutralization Tests; D013997:Time Factors
D000911:Antibodies, Monoclonal; D000939:Epitopes; C096323:rimabotulinumtoxinB; D001905:Botulinum Toxins; D019274:Botulinum Toxins, Type A