High-affinity monoclonal antibodies for botulinum toxin type B
Inventors
Stanker, Larry H. • Scotcher, Miles C. • Cheng, Luisa W. • Hnasko, Robert M. • McGarvey, Jeffery A.
Assignees
US Department of Agriculture USDA
Publication Number
US-9145455-B2
Publication Date
2015-09-29
Expiration Date
2031-09-23
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Abstract
High affinity antibodies for binding epitopes of BoNT/B and hybridomas that produce such antibodies are described. The antibodies may be used in a kit for detecting BoNT/B in a sample.
Core Innovation
The invention describes high affinity monoclonal antibodies (Mab's) that bind to epitopes of botulinum neurotoxin type B (BoNT/B). These antibodies specifically bind to the heavy and light chains of BoNT/B and are produced by hybridomas. A key aspect of the invention is the development of a sandwich ELISA using these Mab's, enabling detection of BoNT/B at concentrations lower than those detectable by the traditional mouse bioassay.
The problem addressed is that while botulinum neurotoxin serotype A (BoNT/A) has been the most studied and characterized, BoNT/B is a significant cause of infant botulism and notable foodborne botulism outbreaks. There has been a need for highly sensitive and specific monoclonal antibodies and assays to detect BoNT/B, similar to those developed for BoNT/A. Existing Mab's for BoNT/A show no cross-reactivity and do not detect BoNT/B, necessitating the development of new Mab's specific for BoNT/B to improve food safety and detection.
The invention also addresses the challenge of detecting BoNT/B in complex matrices like milk without extensive sample preparation. The antibodies exhibit high specificity and neutralization ability in vivo, demonstrating potential for therapeutic use. The Mab's are shown to bind to defined epitopes on the BoNT/B heavy and light chains, and specific antibody combinations allow the construction of sensitive and reproducible sandwich ELISAs suitable for detecting BoNT/B in biological, environmental, and food samples.
Claims Coverage
The patent contains two main inventive features embodied in independent claims relating to a specific monoclonal antibody and its compositions.
Isolated monoclonal antibody produced by specific hybridoma
An isolated and purified monoclonal antibody produced by the continuous hybridoma cell line deposited under accession number ATCC PTA-11872.
Composition comprising the monoclonal antibody
A composition comprising the monoclonal antibody produced by the hybridoma of accession number ATCC PTA-11872.
The independent claims cover the isolated monoclonal antibody specific to BoNT/B produced by a defined hybridoma and compositions containing this antibody, securing protection over the antibody itself and its formulated products.
Stated Advantages
The sandwich ELISA developed using these monoclonal antibodies is approximately 50-fold more sensitive than the traditional mouse bioassay for BoNT/B detection.
The mAb/mAb sandwich ELISA offers greater reagent stability and assay reproducibility compared to assays using monoclonal-polyclonal antibody pairs.
The monoclonal antibodies can detect BoNT/B in milk without the need for defatting or dilution, simplifying sample preparation.
Certain monoclonal antibodies provide complete protection against BoNT/B intoxication in mice, indicating therapeutic potential.
Documented Applications
Use of the monoclonal antibodies in a sandwich ELISA test kit for detecting BoNT/B in buffer and milk samples.
Monitoring and detection of BoNT/B contamination in biological, environmental, and food samples, including milk matrices.
In vivo treatment by administration of monoclonal antibodies to exposed or infected subjects to neutralize BoNT/B toxin effects.
Use of the antibodies as vaccines or therapeutic agents to protect individuals at risk of BoNT/B exposure or infection.
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