High affinity monoclonal antibodies for detection of Shiga toxin 2 (STX2)
Inventors
He, Xiaohua • Stanker, Larry H. • Cheng, Luisa W.
Assignees
US Department of Agriculture USDA
Publication Number
US-9310368-B1
Publication Date
2016-04-12
Expiration Date
2033-09-27
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Abstract
High affinity monoclonal antibodies against Stx2 and hybridomas that produce such antibodies are described. The antibodies may be used in a kit for detecting Stx2 and variants thereof in a sample as well as neutralization of Shiga toxin in vivo.
Core Innovation
The invention describes the production and characterization of high affinity monoclonal antibodies (mAbs) specific to Shiga toxin type 2 (Stx2) and its variants. These mAbs are generated using a genetic toxoid of Stx2a as an immunogen, which preserves the native holotoxin structure and minimizes residual toxicity compared to chemically inactivated toxoids. The invention also provides hybridomas producing these mAbs, including those specific to the shiga toxin 2f variant.
The problem addressed by this invention arises from the challenge in detecting Stx2 in food and biological samples due to low toxin concentrations and complex sample matrices. Existing detection methods such as the Vero cell cytotoxicity assay are time-consuming, require specialized equipment, and need confirmation steps, limiting rapid and sensitive diagnosis. Additionally, commercially available antibodies for Stx2 detection often lack sufficient affinity and specificity, impeding the development of sensitive immunoassays.
This invention solves these problems by providing high affinity mAbs that enable the development of rapid, sensitive, and specific assays such as sandwich ELISAs and lateral flow devices for detecting Stx2 in patient, environmental, and food samples with minimal sample preparation. Furthermore, the mAbs can be used for in vivo neutralization of Stx2 toxicity, offering potential therapeutic or prophylactic applications to protect against infection or exposure.
Claims Coverage
The patent includes three independent claims that cover monoclonal antibodies and their applications related to Shiga toxin 2 detection and complex formation.
Monoclonal antibodies produced by specific hybridoma cell lines
The invention claims monoclonal antibodies produced by continuous hybridoma cell lines deposited under accession numbers PTA-13614 and PTA-13615.
Method for detecting Shiga toxin 2 using monoclonal antibodies
A method comprising incubating a sample with the monoclonal antibody from the specified hybridomas and detecting the antibody-Shiga toxin 2 complex to indicate the presence or absence of the toxin in the sample.
Kit for detecting Shiga toxin 2 in samples
A detection kit comprising a container with the monoclonal antibody produced by the specific hybridomas and instructions for binding and detecting the immunological complex formed with Shiga toxin 2, correlating with its presence or absence in the sample.
The independent claims cover the specific monoclonal antibodies produced by deposited hybridomas, methods to detect Shiga toxin 2 by forming and detecting antibody-toxin complexes, and kits comprising these antibodies with instructions for use in detection, covering key aspects of antibody production, application in detection methods, and provision of detection kits.
Stated Advantages
High affinity monoclonal antibodies improve sensitivity and specificity of immunoassays for Stx2 detection.
Rapid and simple immunoassays (e.g., sandwich ELISA) enable early recognition of STEC contamination in food and clinical samples.
Monoclonal antibodies allow in vivo neutralization of Stx2 toxin, providing protective and therapeutic benefits.
Use of genetically derived toxoid as immunogen preserves holotoxin structure and avoids residual toxicity, enhancing antibody effectiveness.
Monoclonal antibodies accelerate clearance of Stx2 from the bloodstream, reducing disease severity.
Documented Applications
Use of monoclonal antibodies in sandwich ELISA and lateral flow immunoassays for rapid detection of Stx2 in food matrices such as milk, as well as in patient and environmental samples.
Use of monoclonal antibodies for in vivo neutralization and treatment of Stx2 exposure or infection in animal models, with potential therapeutic and prophylactic applications.
Development of kits comprising monoclonal antibodies for detection of Shiga toxin 2 in various samples.
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