Antibodies for detection of colistin-resistance
Inventors
He, Xiaohua • Mavrici, Daniela
Assignees
US Department of Agriculture USDA
Publication Number
US-11161896-B2
Publication Date
2021-11-02
Expiration Date
2039-03-14
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Abstract
Monoclonal antibodies against bacterial proteins that confer resistance to the antibiotic colistin and hybridomas that produce such monoclonal antibodies are described. The monoclonal antibodies have a strong affinity for MCR proteins and may be used in detection methods and kits to detect the presence of MCR proteins including variants thereof in a sample.
Core Innovation
The invention provides monoclonal antibodies and hybridomas that produce monoclonal antibodies with a strong affinity for MCR proteins, which are bacterial proteins conferring resistance to the antibiotic colistin. These monoclonal antibodies can detect the presence of MCR proteins and their variants in various samples, enabling reliable identification of colistin-resistant bacterial strains, including those in biological, environmental, and food samples.
The problem being solved arises from the increasing emergence of multiple drug-resistant bacterial strains, including those resistant to colistin, a last-resort antibiotic. Current detection methods such as PCR suffer from false positives and false negatives, particularly due to genetic variations among mcr gene variants and the presence of inhibitors in biological samples. There exists an urgent need for improved, reliable, and sensitive methods to identify colistin-resistant bacteria promptly to prevent transmission and ensure appropriate treatment.
To address these deficiencies, the invention offers novel monoclonal antibodies specific to MCR proteins and variants, along with methods including immunoassays like sandwich ELISA for sensitive and confident detection. The monoclonal antibodies are produced by specific hybridoma cell lines and may be labeled with various detectable markers. The invention also includes kits comprising these antibodies and instructions for their use in detecting colistin resistance in a wide range of sample types.
Claims Coverage
The patent discloses seven main inventive features derived from independent claims related to monoclonal antibodies, hybridoma cell lines, labeled antibodies, and compositions thereof.
Monoclonal antibodies specific to MCR proteins produced by hybridoma cell lines
Monoclonal antibodies that specifically react with at least one mobilized colistin resistance (MCR) protein, produced by hybridoma cell lines having ATCC Patent Deposit Designation No. PTA-125013 or PTA-125014.
Isolated and purified monoclonal antibodies or fragments thereof
The monoclonal antibodies or their fragments are isolated and purified to allow high specificity and affinity for MCR proteins.
Chimeric monoclonal antibodies
Chimeric monoclonal antibodies or fragments thereof are provided, which incorporate antibody portions from different species or antibody classes to achieve desired immunologic properties.
Labeled monoclonal antibodies with detectable markers
Monoclonal antibodies or fragments thereof further comprise labels selected from enzyme labels, radioisotopic labels, non-radioactive isotopic labels, chromogenic labels, fluorescent labels, chemiluminescent labels, or combinations thereof to facilitate detection.
Hybridoma cell lines producing the described monoclonal antibodies
Hybridoma cell lines that produce the monoclonal antibodies or fragments thereof that react specifically with MCR proteins, particularly those having ATCC Patent Deposit Designation No. PTA-125013 or PTA-125014.
Composition comprising the monoclonal antibodies
Compositions comprising the monoclonal antibody that reacts specifically with MCR proteins.
The claimed invention encompasses monoclonal antibodies specific to MCR proteins and their production via defined hybridoma cell lines, including isolated, labeled, and chimeric forms, as well as compositions comprising these antibodies. These features collectively enable sensitive and specific detection of colistin resistance.
Stated Advantages
Provides a collection of novel monoclonal antibodies to detect and confirm the presence of MCR proteins and variants with high sensitivity and confidence.
Offers novel hybridoma cell lines that produce monoclonal antibodies with specificity to MCR proteins, enabling consistent detection.
Facilitates diagnostic tests using these antibodies individually or in combinations to detect MCR proteins in various sample types such as animals, patients, environment, and food processing stages.
Provides kits for detecting MCR proteins and variants in samples, enhancing ease of use and application in different settings.
Documented Applications
Detection and confirmation of colistin-resistant bacterial strains in samples such as aqueous, biological, environmental, fresh food, food products, and processed food products.
Screening of meat products including ground beef, chicken, and pork for contamination with colistin-resistant bacteria carrying mcr genes.
Use in epidemiological surveillance programs targeting colistin-resistant bacteria from human and animal sources.
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