Development of an innovative immunoassay for CP4EPSPS and Cry1AB genetically modified protein detection and quantification

Abstract

An innovative immunoassay, called enzyme-linked immunoabsorbant assay (ELISA) Reverse, based on a new conformation of the solid phase, was developed. The solid support was expressly designed to be immersed directly in liquid samples to detect the presence of protein targets. Its application is proposed in those cases where a large number of samples have to be screened simultaneously or when the simultaneous detection of different proteins is required. As a first application, a quantitative immunoassay for Cry1AB protein in genetically modified maize was optimized. The method was tested using genetically modified organism concentrations from 0.1 to 2.0%. The limit of detection and limit of quantitation of the method were determined as 0.0056 and 0.0168 (expressed as the percentage of genetically modified organisms content), respectively. A qualitative multiplex assay to assess the presence of two genetically modified proteins simultaneously was also established for the case of the Cry1AB and the CP4EPSPS (5-enolpyruvylshikimate-3-phosphate synthase) present in genetically modified maize and soy, respectively.

Keywords:

• Genetically modified organisms (GMOs)
• immunoassay
• enzyme-linked immunoabsorbant assay (ELISA) Reverse
• Cry1AB
• CP4EPSPS

Acknowledgements

The collaboration between DG-Joint Research Centre (JRC) and Molise University was enhanced by the ENEA-Innovation Relay Centre IRIDE as part of the IRC NETWORK created by European Commission.