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-8900824-B1
Publication Date
2014-12-02
Expiration Date
2031-09-23
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
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 specifically bind to epitopes of botulinum neurotoxin type B (BoNT/B), including both the heavy and light chains of the toxin. These monoclonal antibodies are IgG1 subclass with kappa light chains, produced by hybridomas, and characterized for their binding specificity and application in rapid immunoassay formats. The development includes methods for detecting BoNT/B using these antibodies in sensitive sandwich ELISA assays with lower detection limits than established mouse bioassays.
The invention addresses the challenge of detecting BoNT/B in various samples, resolving the need for a specific and sensitive assay for BoNT/B, unlike prior development focused primarily on BoNT/A. While BoNT/B is less frequently associated with foodborne botulism cases, it remains a significant threat, especially given its role in infant botulism and historic outbreaks. Existing assays for BoNT/A did not cross-react with BoNT/B, highlighting the need for novel monoclonal antibodies that can specifically detect BoNT/B with high sensitivity and facilitate rapid immunological detection approaches.
The invention further encompasses the application of these monoclonal antibodies in sandwich ELISA kits that detect BoNT/B in buffer and complex food matrices such as milk, with minimal sample preparation. The antibodies can capture and detect BoNT/B at concentrations undetectable by traditional bioassays and can neutralize BoNT/B toxicity in vivo, providing potential therapeutic or prophylactic uses. The invention also includes kits and methods for detecting BoNT/B in environmental, biological, or food samples using these antibodies.
Claims Coverage
The patent contains several independent claims focusing on isolated monoclonal antibodies, compositions comprising these antibodies, methods of detecting BoNT/B, sandwich immunoassays, kits, and methods of capturing BoNT/B.
Isolated monoclonal antibody produced by continuous hybridoma cell line
An isolated and purified monoclonal antibody produced by the continuous hybridoma cell line having deposit accession number ATCC PTA-11871.
Composition comprising the monoclonal antibody
A composition comprising the monoclonal antibody produced by the hybridoma cell line ATCC PTA-11871.
Method for detecting BoNT/B using monoclonal antibody
A method for detecting BoNT/B by incubating a sample with the monoclonal antibody from cell line ATCC PTA-11871 to form an antibody-BoNT/B complex and detecting the presence or absence of the complex to indicate BoNT/B presence.
Sandwich immunoassay for BoNT/B detection
The detection method consists essentially of a sandwich immunoassay format using the monoclonal antibody.
Kit comprising monoclonal antibody and instructions for BoNT/B detection
A kit for detecting BoNT/B including a container with the monoclonal antibody from hybridoma ATCC PTA-11871 and instructions for using the antibody to bind BoNT/B and detect the immunological complex.
Method for detecting BoNT/B in various sample types
A method according to claim 3 where the sample is aqueous, biological, environmental, or a food product.
Method for capturing BoNT/B from a sample
A method for capturing BoNT/B by contacting the sample with the monoclonal antibody from claim 1 and isolating the complex formed between BoNT/B and the antibody.
These claims cover the production of a specific monoclonal antibody against BoNT/B, compositions containing it, methods and kits for detection of BoNT/B using immunoassays including sandwich ELISA formats, and the capture of BoNT/B from samples using the antibody, emphasizing the specificity and practical applications of the monoclonal antibody derived from deposited hybridoma cell lines.
Stated Advantages
The antibodies exhibit high affinity and specificity to BoNT/B, allowing detection at concentrations lower than those detectable by mouse bioassays.
The sandwich ELISA developed using these monoclonal antibodies enables rapid, sensitive, and reproducible detection of BoNT/B in complex matrices including various types of milk without need for sample defatting or dilution.
Monoclonal antibodies, particularly MCS6-27 and MCS92-32-10-1, provide potential for in vivo treatment or prophylaxis against BoNT/B exposure, demonstrating protective efficacy in a mouse model.
Use of monoclonal-monoclonal antibody pairs in assays ensures unlimited reagent availability and greater assay stability and reproducibility compared to monoclonal-polyclonal pairs.
Documented Applications
Use in kits for detecting BoNT/B in samples including biological, environmental, and food samples.
Application of sandwich ELISA assays to detect BoNT/B in buffer and food matrices such as skim, 2% fat, and whole milk with minimal sample preparation.
Use of monoclonal antibodies for in vivo neutralization and therapeutic treatment of Clostridium botulinum neurotoxin type B intoxication.
Interested in licensing this patent?