title: Production of a neutralizing mouse-human chimeric antibody against botulinum neurotoxin serotype E.
abstract: A mouse-human chimeric antibody that can neutralize botulinum neurotoxin serotype E (BoNT/E) was developed. Variable regions of heavy and light chains obtained using a mouse hybridoma clone (E9-4) cDNA, which was selected on the basis of neutralizing activity against BoNT/E, were fused with the upstream regions of the constant counterparts of human kappa light and gamma 1 heavy chain genes, respectively. CHO-DG44 cells were transfected with these plasmids and a mouse-human chimeric antibody (EC94) was purified to examine binding and neutralizing activity against BoNT/E. EC94 exhibited the same levels of binding activities against BoNT/E as those of a parent mouse monoclonal antibody and neutralized more than 4,000 LD(50)/mg antibody. This chimeric antibody seems to be a useful candidate for infant botulism in which the use of passive immunotherapy is not planned so as to avoid serious events such as anaphylactic shock. We designed shuffling chimeric antibodies with replacement of V(H) or V(L) of EC94 with that of a chimeric antibody (AC24) that possessed neutralizing activity against BoNT/A. These shuffling antibodies did not exhibit neutralizing activity against either BoNT/E or BoNT/A.
authors: Department of Veterinary Science, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Osaka 598-8531, Japan. mukamoto@vet.osakafu-u.ac.jp
authors: Masafumi
authors: M
authors: Mukamoto
authors: Hiroaki
authors: H
authors: Maeda
authors: Tomoko
authors: T
authors: Kohda
authors: Chikateru
authors: C
authors: Nozaki
authors: Motohide
authors: M
authors: Takahashi
authors: Shunji
authors: S
authors: Kozaki
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D057134:Antibodies, Neutralizing / Q000096:biosynthesis* / Q000276:immunology; D001905:Botulinum Toxins / Q000276:immunology*; D001906:Botulism / Q000276:immunology / Q000517:prevention & control*; D002478:Cells, Cultured; D002678:Chimera; D003014:Clostridium botulinum / Q000276:immunology / Q000472:pathogenicity; D047970:Clostridium botulinum type E / Q000276:immunology*; D046972:Clostridium butyricum / Q000276:immunology / Q000472:pathogenicity; D006801:Humans; D006825:Hybridomas; D007142:Immunoglobulin gamma-Chains / Q000235:genetics / Q000276:immunology; D007145:Immunoglobulin kappa-Chains / Q000235:genetics / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009498:Neurotoxins / Q000276:immunology; D009500:Neutralization Tests; D011993:Recombinant Fusion Proteins
chemical_list: D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D007142:Immunoglobulin gamma-Chains; D007145:Immunoglobulin kappa-Chains; D009498:Neurotoxins; D011993:Recombinant Fusion Proteins; D001905:Botulinum Toxins; C096326:botulinum toxin type E