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Journal Article | Research Support, U.S. Gov't, Non-P.H.S.

Antibody mapping to domains of botulinum neurotoxin serotype A in the complexed and uncomplexed forms.

F Chen, G M Kuziemko, P Amersdorfer, C Wong, J D Marks, R C Stevens
F Chen
Graduate Group in Biophysics, University of California, Berkeley 94720, USA.
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G M Kuziemko
Graduate Group in Biophysics, University of California, Berkeley 94720, USA.
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P Amersdorfer
Graduate Group in Biophysics, University of California, Berkeley 94720, USA.
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C Wong
Graduate Group in Biophysics, University of California, Berkeley 94720, USA.
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J D Marks
Graduate Group in Biophysics, University of California, Berkeley 94720, USA.
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R C Stevens
Graduate Group in Biophysics, University of California, Berkeley 94720, USA.
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ABSTRACT

The domain organization of the botulinum neurotoxin serotype A was studied by using antibody mapping of 44 monoclonal single-chain variable fragments. The analysis was carried out on (i) the individual domains of botulinum neurotoxin holotoxin (binding, translocation, and catalytic), (ii) botulinum neurotoxin holotoxin, (iii) the botulinum neurotoxin holotoxin in complex with the nontoxic portion, and (iv) botulinum neurotoxin holotoxin and nontoxic portion of the complex recombined in vitro. All 44 antibodies mapped to individual domains of botulinum neurotoxin. Forty of the 44 single-chain variable fragments bound the botulinum neurotoxin holotoxin relative to the isolated domains, suggesting that 4 epitopes are covered when the individual domains are in the holotoxin form. Only 20 of the antibodies showed a positive reaction to the toxin while in complex with the nontoxic portion. All of the covered epitopes were mapped to the binding domain of botulinum neurotoxin, which suggested that the binding domain is in direct contact with the nontoxic portion in the complex. Based on the antibody mapping to the different domains of the botulinum neurotoxin holotoxin and the entire complex, a model of the botulinum neurotoxin complex is proposed.

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Antibody mapping to domains of botulinum neurotoxin serotype A in the complexed and uncomplexed forms.
F Chen, G M Kuziemko, P Amersdorfer, C Wong, J D Marks, R C Stevens
Infection and Immunity May 1997, 65 (5) 1626-1630; DOI:

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Antibody mapping to domains of botulinum neurotoxin serotype A in the complexed and uncomplexed forms.
F Chen, G M Kuziemko, P Amersdorfer, C Wong, J D Marks, R C Stevens
Infection and Immunity May 1997, 65 (5) 1626-1630; DOI:
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