Quantitative ELISA validation for GE crop expression

AGRO 53

Valerie A. Korjagin, vakorjagin@dow.com, Guomin Shan, gshan@dow.com, Shawna K. Embrey, and Angelea D. Thomas. Regulatory Laboratories, Dow AgroSciences LLC, 9330 Zionsville Rd, Indianapolis, IN 46268
Enzyme linked immunosorbent assay (ELISA) has become a very widely accepted analytical technique used in the quantitative detection of proteins of interest in genetically enhanced (GE) crops. The use of ELISA is very appealing due to the ease of use, quick turn around time, low cost per sample analysis and compatibility with high throughput automation. To ensure the integrity of the data, a validation of an ELISA is needed and defines important parameters including precision (repeatability/reproducibility), accuracy (recovery and false negative/positive rates), ruggedness, matrix effects and interfering substances. In addition to these, sensitivity, including limit of detection and quantitation and specificity of the assay are also determined. We have validated multiple ELISAs for the determination of Bt proteins such as Cry1F and Cry1Ac in various plant tissues. The validation results will be discussed. A comprehensive validation process including acceptance criteria will be discussed as well.