ABSTRACT
Rapid development and global cultivation of genetically modified (GM) crops necessitated the use of analytical tools that detect GM crops throughout the product life cycle. Antibody-based immunoassays such as enzyme-linked immunosorbent assays (ELISA) are instrumental in detecting protein expression in transgenic plants. These analytical tools are used throughout development, regulatory registration, commercialization, and stewardship of biotech products. Here we describe the Cry1F ELISA analytical method protocol, data generation and review, and troubleshooting of technical challenges.
Subject(s)
Bacterial Proteins/metabolism , Crops, Agricultural/metabolism , Plants, Genetically Modified/metabolism , Bacterial Proteins/genetics , Bacterial Proteins/immunology , Crops, Agricultural/genetics , Enzyme-Linked Immunosorbent Assay/instrumentation , Enzyme-Linked Immunosorbent Assay/methods , Plants, Genetically Modified/genetics , Transgenes/geneticsABSTRACT
The concentration of transgene products (proteins and double-stranded RNA) in genetically modified (GM) crop tissues is measured to support food, feed, and environmental risk assessments. Measurement of transgene product concentrations in breeding stacks of previously assessed and approved GM events is required by many regulatory authorities to evaluate unexpected transgene interactions that might affect expression. Research was conducted to determine how well concentrations of transgene products in single GM events predict levels in breeding stacks composed of these events. The concentrations of transgene products were compared between GM maize, soybean, and cotton breeding stacks (MON-87427 × MON-89034 × DAS-Ø15Ø7-1 × MON-87411 × DAS-59122-7 × DAS-40278-9 corn, DAS-81419-2 × DAS-44406-6 soybean, and DAS-21023-5 × DAS-24236-5 × SYN-IR102-7 × MON-88913-8 × DAS-81910-7 cotton) and their component single events (MON-87427, MON-89034, DAS-Ø15Ø7-1, MON-87411, DAS-59122-7, and DAS-40278-9 corn, DAS-81419-2, and DAS-44406-6 soybean, and DAS-21023-5, DAS-24236-5, SYN-IR102-7, MON-88913-8, and DAS-81910-7 cotton). Comparisons were made within a crop and transgene product across plant tissue types and were also made across transgene products in each breeding stack for grain/seed. Scatter plots were generated comparing expression in the stacks to their component events, and the percent of variability accounted for by the line of identity (y = x) was calculated (coefficient of identity, I2). Results support transgene concentrations in single events predicting similar concentrations in breeding stacks containing the single events. Therefore, food, feed, and environmental risk assessments based on concentrations of transgene products in single GM events are generally applicable to breeding stacks composed of these events.