Hybridomas producing highly specific monoclonal antibodies to detect mycobacterium avium subspecies paratuberculosis
Inventors
Assignees
US Department of Agriculture USDA
Publication Number
US-8658386-B1
Publication Date
2014-02-25
Expiration Date
2028-12-17
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Abstract
Hybridoma cell lines which produce and secrete monoclonal antibodies which selectively bind to Mycobacterium avium subspecies paratuberculosis have been produced. Cells of M. avium subspecies paratuberculosis in biological samples may be detected and quantified by contacting the sample with the antibodies to form a M. avium subspecies paratuberculosis/antibody immunocomplex when M. avium subspecies paratuberculosis is present, which immunocomplex may then be detected. The monoclonal antibodies also may be incorporated into kits for the detection and quantification of M. avium subspecies paratuberculosis.
Core Innovation
The invention relates to hybridoma cell lines that produce monoclonal antibodies which selectively bind to Mycobacterium avium subspecies paratuberculosis (MAP). These monoclonal antibodies enable the detection and quantification of MAP in biological samples by forming immunocomplexes when MAP is present, which can then be detected through immunoassays. The antibodies may also be incorporated into diagnostic kits for MAP detection and quantification.
The problem addressed is the lack of specific, antigen-based detection reagents for MAP despite the significant economic impact of Johne's disease caused by MAP, especially in the dairy industry. Prior art presents very few reports on MAP-specific monoclonal antibodies, and existing detection methods have low sensitivity and specificity. There is a vital need for improved diagnostic reagents such as monoclonal antibodies for MAP to enhance research and diagnostic capabilities related to Johne's disease.
Claims Coverage
The patent claims cover one independent claim directed to a hybridoma cell line and one independent claim directed to a method for detecting Mycobacterium avium subspecies paratuberculosis.
Hybridoma cell line producing monoclonal antibodies selectively binding to Mycobacterium avium subspecies paratuberculosis
The hybridoma cell line designated 12C9 (ATCC PTA-10623) produces and secretes monoclonal antibodies that selectively bind to Mycobacterium avium subspecies paratuberculosis.
Method for detecting Mycobacterium avium subspecies paratuberculosis in a biological sample using monoclonal antibodies from hybridoma 12C9
A method comprising providing a biological sample, subjecting it to an immunosorbent assay using monoclonal antibodies produced by hybridoma cell line 12C9 that selectively bind MAP, forming an immunocomplex if MAP is present, and detecting this immunocomplex, whereby detection indicates the presence of MAP in the sample. The sample can be animal tissue or fluids.
The claimed inventions include a specific hybridoma cell line producing MAP-selective monoclonal antibodies and methods employing these antibodies for detecting MAP in biological samples through immunoassays, covering both the biological material and detection process.
Stated Advantages
Provides hybridoma cell lines that produce high affinity monoclonal antibodies which selectively bind to Mycobacterium avium subspecies paratuberculosis.
Enables rapid and accurate detection and quantification of Mycobacterium avium subspecies paratuberculosis in biological samples.
Facilitates purification and concentration of Mycobacterium avium subspecies paratuberculosis from environmental or biological samples.
Allows incorporation of monoclonal antibodies into diagnostic kits for specific detection and quantification of the microorganism.
Documented Applications
Use in immunoassay methods such as RIA, FIA, ELISA, competitive inhibition ELISA, and double-antibody sandwich ELISA for detecting and quantifying Mycobacterium avium subspecies paratuberculosis in animal tissue and fluids.
Purification and concentration of Mycobacterium avium subspecies paratuberculosis from environmental or biological samples using monoclonal antibodies bound to a solid support or matrix.
Incorporation into diagnostic kits comprising antibodies, detection means, and solid phase supports for MAP antigens.
Use in histopathological examination of infected tissues, particularly for detecting MAP in intestinal lamina propria.
Application in research involving cell biology, pathogenesis, macrophage-pathogen interactions, and immunofluorescence microscopy.
Potential incorporation into sensors such as solid phase electronic devices for MAP detection in samples.
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