Epitope characterization of sero-specific monoclonal antibody to Clostridium botulinum neurotoxin type A.

Authors

Cindi R Corbett, National Microbiology Laboratory, 1015 Arlington Street, Winnipeg, MB; Department of Medical Microbiology and Infectious Diseases, University of Manitoba
Erin Ballegeer, National Microbiology Laboratory, 1015 Arlington Street, Winnipeg, MB
Kelly A Weedmark, National Microbiology Laboratory, 1015 Arlington Street, Winnipeg, MB
M D Elias, Center for Research on Bioterrorism and Biodefense, Department of Medicine, Jefferson Medical CollegeFollow
Fetweh H Al-Saleem, Center for Research on Bioterrorism and Biodefense, Department of Medicine, Jefferson Medical CollegeFollow
Denise M Ancharski, Center for Research on Bioterrorism and Biodefense, Department of Medicine, Jefferson Medical CollegeFollow
Lance L Simpson, Center for Research on Bioterrorism and Biodefense, Department of Medicine, Jefferson Medical CollegeFollow
Jody D Berry, National Microbiology Laboratory, 1015 Arlington Street, Winnipeg, MB; Department of Medical Microbiology and Infectious Diseases, University of Manitoba; Department of Immunology, Faculty of Medicine, University of Manitoba

Document Type

Article

Publication Date

12-1-2011

Comments

This article has been peer reviewed and is published in Hybridoma December 2011, 30(6): 503-510. The published version is available at DOI: doi:10.1089/hyb.2011.0032. ©Mary Ann Liebert, Inc

Abstract

Botulinum neurotoxins (BoNTs) are extremely potent toxins that can contaminate foods and are a public health concern. Anti-BoNT antibodies have been described that are capable of detecting BoNTs; however there still exists a need for accurate and sensitive detection capabilities for BoNTs. Herein, we describe the characterization of a panel of eight monoclonal antibodies (MAbs) generated to the non-toxic receptor-binding domain of BoNT/A (H(C)50/A) developed using a high-throughput screening approach. In two independent hybridoma fusions, two groups of four IgG MAbs were developed against recombinant H(C)50/A. Of these eight, only a single MAb, F90G5-3, bound to the whole BoNT/A protein and was characterized further. The F90G5-3 MAb slightly prolonged time to death in an in vivo mouse bioassay and was mapped by pepscan to a peptide epitope in the N-terminal subdomain of H(C)50/A (H(CN)25/A) comprising amino acid residues (985)WTLQDTQEIKQRVVF(999), an epitope that is highly immunoreactive in humans. Furthermore, we demonstrate that F90G5-3 binds BoNT/A with nanomolar efficiency. Together, our results indicate that F90G5-3 is of potential value as a diagnostic immunoreagent for BoNT/A capture assay development and bio-forensic analysis.

Recommended Citation

Corbett, Cindi R; Ballegeer, Erin; Weedmark, Kelly A; Elias, M D; Al-Saleem, Fetweh H; Ancharski, Denise M; Simpson, Lance L; and Berry, Jody D, "Epitope characterization of sero-specific monoclonal antibody to Clostridium botulinum neurotoxin type A." (2011). Department of Medicine Faculty Papers. Paper 73.
https://jdc.jefferson.edu/medfp/73

PubMed ID

22149274