Monoclonal antibodies detection methods for enzymes that confer resistance to 2,4-dichlorophenoxyacetic acid in plants

• US 8,460,891 B2
• Filed: 06/02/2011
• Issued: 06/11/2013
• Est. Priority Date: 06/04/2010
• Status: Active Grant

• Current Assignee: Corteva Agriscience LLC (Corteva, Inc.)
• Original Assignee: Dow Agrosciences LLC (Corteva, Inc.)
• Inventors: Shan, Guomin, Lin, Gaofeng, Sosa, Marcelo, Smith Drake, Joelene K.
• Primary Examiner(s): FOSTER, CHRISTINE E
• Application Number:
  US13/151,389
• Publication Number:
  US 20110300556A1
• Time in Patent Office: 740 Days
• Field of Search: None
• US Class Current: 435/7.4
• CPC Class Codes: C07K 16/40 against enzymes
• CPC Class Codes: G01N 33/573 for enzymes or isoenzymes

Abstract

Described herein are monoclonal antibodies and methods useful for determining and quantitating the presence of an aryloxyalkanoate dioxygenase enzyme.

Claims

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.