Production of a Neutralizing Mouse-Human Chimeric Antibody against Botulinum Neurotoxin Serotype E

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  • Mukamoto Masafumi
    Department of Veterinary Science, Graduate School of Life and Environmental Sciences, Osaka Prefecture University
  • Maeda Hiroaki
    The Chemo-Sero-Therapeutic Research Institute (KAKETSUKEN), Kikuchi Research Center
  • Kohda Tomoko
    Department of Veterinary Science, Graduate School of Life and Environmental Sciences, Osaka Prefecture University
  • Nozaki Chikateru
    The Chemo-Sero-Therapeutic Research Institute (KAKETSUKEN), Kikuchi Research Center
  • Takahashi Motohide
    Department of Bacteriology II, National Institute of Infectious Diseases
  • Kozaki Shunji
    Department of Veterinary Science, Graduate School of Life and Environmental Sciences, Osaka Prefecture University

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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 LD50/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 VH or VL 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.

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