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http://dx.doi.org/10.5762/KAIS.2017.18.1.295

Production and Characterization of a Recombinant Antibody Neutralizing Botulinum Neurotoxin A  

Park, Hong-Gyu (R&D Department, ATGC Co., LTD.)
Choi, Mieyoung (Department of BT-Convergent Pharmaceutical Engineering, Sun Moon University)
Publication Information
Journal of the Korea Academia-Industrial cooperation Society / v.18, no.1, 2017 , pp. 295-301 More about this Journal
Abstract
Botulinum neurotoxin (BoNT/A) is a neurotoxin that selectively attacks the peripheral cholinergic nerve endings. It is produced by Gram -positive, endospore-forming strict anaerobic bacteria, Clostridium botulinum. Since BoNT/A could be a biothreat agent, as well as a contaminator of food and water supplies, the development of sensitive assays for toxin detection and potent antitoxin for the treatment of intoxication is necessary. In this study, for the purpose of producing monoclonal antibodies (mAbs) that are capable of neutralizing Botulinum neurotoxin type A (BoNT/A), scFv (single-chain variable domain fragment) libraries from the rabbit antisera against BoNT/A was fused to a human IgG. The resulting recombinant scFvIgG antibody protein was expressed in stable cell lines and was purified using a protein A affinity chromatography. The efficacy of scFvIgG mAb was confirmed by ELISA and was evaluated for the neutralization of BoNT/A in vivo. Such an in vivo toxin neutralization assay was performed using mice. Although scFvIgG antibody proteins (10 ug) failed to fully protect the mice challenged with BoNT/A (100,000 $LD_{50}$), it significantly prolonged the survival time. These results suggest that scFvIgG mAb may be capable of neutralizing BoNT/A single-chain variable domain fragment.
Keywords
Botulinum neurotoxin type A(BoNT/A); ELISA(enzyme-linked immunosorbent assay); in vivo toxin neutralization assay; monoclonal antibody(mAb); scFv(single-chain variable domain fragment);
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