title: Characterization of the monoclonal antibody response to botulinum neurotoxin type A in the complexed and uncomplexed forms.
abstract: Clostridium botulinum produces large complex toxins, which include botulinum  neurotoxin (BoNT) and auxiliary non-toxic proteins. We prepared monoclonal antibodies (mAbs) from  mice that were immunized several times with BoNT/A after basal immunization with toxoid. We then  examined the reactivities of these mAbs to BoNT and toxoid and showed that some mAbs reacted to  only BoNT. This result indicates that the antigenicity of BoNT/A partially disappeared with  formalin treatment. Some mAbs that specifically recognized either BoNT/A1 or BoNT/A2 were  considered useful as detection antibodies specific for the BoNT/A subtype. Results of a  neutralizing test with mAbs against either BoNT/A1 or BoNT/A2 showed that neutralizing antibody  recognition sites were present in the light chain, heavy chain (N-terminal half), and heavy chain  (C-terminal half) domains. Investigation of the different binding capabilities of the mAbs to BoNT  and the complex toxin by immunoprecipitation suggested that the light chain of BoNT is exposed at  the molecular surface of the complex toxin since there was no difference in the binding of light  chain-specific mAb to BoNT and the complex toxin. The heavy chain is related to BoNT binding to  non-toxic components, because the reactivity of the heavy chain to some mAbs was influenced by  non-toxic components.
authors.affiliation: Department of Veterinary Science, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Osaka 598-8531, Japan.
authors.forename: Haiyang
authors.initials: H
authors.lastname: Zhao
authors.forename: Keiji
authors.initials: K
authors.lastname: Nakamura
authors.forename: Tomoko
authors.initials: T
authors.lastname: Kohda
authors.forename: Masafumi
authors.initials: M
authors.lastname: Mukamoto
authors.forename: Shunji
authors.initials: S
authors.lastname: Kozaki
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology* / Q000302:isolation & purification; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D057134:Antibodies, Neutralizing / Q000276:immunology* / Q000302:isolation & purification; D000992:Antitoxins / Q000276:immunology* / Q000302:isolation & purification; D019274:Botulinum Toxins, Type A / Q000276:immunology*; D000939:Epitopes / Q000276:immunology; D047468:Immunoprecipitation; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D016415:Sequence Alignment
chemical_list: D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D000992:Antitoxins; D000939:Epitopes; D019274:Botulinum Toxins, Type A