Compositions and methods for detection of wild type cry protein and a hybrid cry protein
AssigneeSyngenta Participations AG
InventorsSimone Cummings, Julie Smith, Magda Foege
Life of this patent
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Abstract
The invention relates generally to immunoassays, and more particularly to monoclonal antibodies and immunoassays for the differential detection and quantitation of a wild-type crystal protein, such as a wild-type-Cry1Ab, from Bacillus thuringiensis and hybrid crystal proteins, which comprise all or a significant portion of the wild-type Cry protein in complex biological samples comprising both the wild-type Cry protein and one or more of the hybrid Cry proteins.
Claims16
A composition comprising a first monoclonal MAb58 antibody produced by a hybridoma cell line deposited as ATCC Accession No. PTA-122985 and a second monoclonal antibody H04MAb70 produced by a hybridoma cell line deposited as ATCC Accession No. PTA-122984, wherein the first and second antibody that function together to specifically detect or quantitate a Cry1A.105 protein comprising SEQ ID NO:4 in an immunoassay of a biological sample comprising the Cry1A.105 protein and a Cry1Ab protein comprising SEQ ID NO:1, wherein the first antibody and the second antibody individually are capable of binding to both the Cry1Ab protein and the Cry1A.105 protein but that function together in the immunoassay to specifically detect or quantitate the Cry1A.105 protein and not the Cry1Ab protein.
A monoclonal antibody produced by a hybridoma deposited as ATCC Accession No. PTA-122985.
An immunoassay method to detect or quantitate a Cry1A.105 protein comprising SEQ ID NO:4 and a Cry1Ab protein comprising SEQ ID NO:1 in a biological sample from a transgenic corn plant comprising the Cry1Ab protein and the Cry1A.105 protein, the method comprising:
- (a) coating a first solid surface with a first antibody that is MAb58 produced by a hybridoma cell line deposited as ATCC Accession No. PTA-122985 that binds the Cry1A.105 protein but does not bind the Cry1Ab protein and coating a second solid surface with a second antibody that is 87AB1.1 that binds the Cry1Ab protein;
- (b) contacting the biological sample with the MAb58 antibody under conditions effective to allow the formation of a Cry1A.105-MAb58 antibody complex but does not allow the formation of a Cry1Ab-MAb58 antibody complex, resulting in a Cry1A.105-depleted biological sample;
- (c) removing the Cry1A.105-depleted biological sample and contacting the Cry1A.105-depleted biological sample with the 87AB1.1 antibody under conditions effective to allow the formation of a Cry1Ab protein-87AB1.1 antibody complex;
- (d) detecting or quantitating the Cry1A.105 protein complex on the first solid surface;
- and (e) detecting or quantitating the Cry1Ab protein on the second solid surface.
Dependencies are inferred from the claim text.