Monoclonal antibodies detection methods for enzymes that confer resistance to 2,4-dichlorophenoxyacetic acid in plants
View Patent ↗Described herein are monoclonal antibodies and methods useful for determining and quantitating the presence of an aryloxyalkanoate dioxygenase enzyme.
1. An isolated monoclonal antibody that specifically binds to aryloxyalkanoate dioxygenase-12 enzyme (AAD-12), wherein the monoclonal antibody is produced by a hybridoma having an American Type Culture Collection (ATCC) Accession Number selected from the group consisting of PTA-10919, PTA-10920, PTA-10921, PTA-10922, and PTA-10923.
2. The monoclonal antibody of claim 1 produced by the hybridoma having ATCC Accession Number PTA-10919.
3. The monoclonal antibody of claim 1 produced by the hybridoma having ATCC Accession Number PTA-10920.
4. The monoclonal antibody of claim 1 produced by the hybridoma having ATCC Accession Number PTA-10921.
5. The monoclonal antibody of claim 1 produced by the hybridoma having ATCC Accession Number PTA-10922.
6. The monoclonal antibody of claim 1 produced by the hybridoma having ATCC Accession Number PTA-10923.
7. A hybridoma cell line that produces a monoclonal antibody of claim 1 that is on deposit with the American Type Culture Collection (ATCC) under Accession Numbers selected from the group consisting of PTA-10919, PTA-10920, PTA-10921, PTA-10922, and PTA-10923.
8. The hybridoma cell line of claim 7 deposited under ATCC Accession Number PTA-10919.
9. The hybridoma cell line of claim 7 deposited under ATCC Accession Number PTA-10920.
10. The hybridoma cell line of claim 7 deposited under ATCC Accession Number PTA-10921.
11. The hybridoma cell line of claim 7 deposited under ATCC Accession Number PTA-10922.
12. The hybridoma cell line of claim 7 deposited under ATCC Accession Number PTA-10923.
13. A method for identifying the presence of AAD-12 enzyme comprising:
a) immobilizing a first monoclonal antibody of claim 1 onto an assay surface, and then washing said assay surface;
b) contacting said assay surface with a liquid suspected of containing AAD-12 for a period of time sufficient to allow binding, and then washing said assay surface;
c) contacting said assay surface with a different second antibody of claim 1 conjugated to a reporting group for a period of time sufficient to allow binding of said second conjugated monoclonal antibody, and then washing said assay surface; and,
d) detecting the presence or absence of said reporting group, wherein the presence of said reporting group indicates the presence of AAD-12 in the liquid.
14. A method for the quantitative determination of AAD-12 enzyme comprising:
a) immobilizing an AAD-12 antiserum onto an assay surface;
b) contacting said assay surface with a liquid suspected of containing AAD-12 for a period of time sufficient to allow binding then washing said assay surface;
c) contacting said assay surface with a monoclonal antibody of claim 1 conjugated to a reporting group for a period of time sufficient to allow binding of said second conjugated monoclonal antibody, and then washing said assay surface; and,
d) quantitating the presence of said reporting group by comparison to a calibration curve, wherein the presence of said reporting group indicates the presence of AAD-12 in the liquid.
15. The method of claim 14 wherein the conjugated monoclonal antibody is produced by the hybridoma having ATCC Accession Number PTA-10920.