A transgenic maize line has been genetically modified to express aryloxyalkanoate dioxygenase protein (AAD-1) providing tolerance to 2,4-dichlorophenoxyacetic acid (2,4-D) and aryloxyphenoxypropionate (fop) herbicides. An Enzyme Linked-Immunosorbent Assay (ELISA) has been developed to specifically quantitiate the AAD-1 expression in maize tissues. The resulting assay uses two specific monoclonal antibodies in a sandwich ELISA format and was validated for its sensitivity, specificity, accuracy, precision and ruggedness. The validation results demonstrated that it is specific to AAD-1 with no cross-reactivity to other transgenic engineered proteins. The AAD-1 protein extracted from various maize tissues can be quantitated from a 1 � 32 ng/mL calibration curve with acceptable accuracy and precision. The validated LOD (limit of detection) and lower LOQ (limit of quantitation) were 0.20 and 0.40 ng/mg dry weight, respectively. The results demonstrate that the assay is specific, sensitive, and accurate for AAD-1 quantitation in maize tissues.

Development and validation of a sensitive and specific immunoassay for the detection of aryloxyalkanoate dioxygenase (AAD-1) protein in maize tissues

246th National Meeting (2013)

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