Development and validation of a sensitive enzyme immunoassay for surveillance of Cry1Ab toxin in bovine blood plasma of cows fed Bt-maize (MON810) |
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Authors: | Paul Vijay Steinke Kerstin Meyer Heinrich H D |
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Institution: | a Physiology Weihenstephan, Technical University Munich, Weihenstephaner Berg 3, 85350 Freising, Germany b Bavarian State Research Center for Agriculture, Institute for Animal Nutrition and Feed Management, Prof.-Duerrwaechter-Platz 3, 85586 Poing, Germany |
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Abstract: | The increasing global adoption of genetically modified (GM) plant derivatives in animal feed has provoked a strong demand for an appropriate detection method to evaluate the existence of transgenic protein in animal tissues and animal by-products derived from GM plant fed animals. A highly specific and sensitive sandwich enzyme immunoassay for the surveillance of transgenic Cry1Ab protein from Bt-maize in the blood plasma of cows fed on Bt-maize was developed and validated according to the criteria of EU-Decision 2002/657/EC. The sandwich assay is based on immuno-affinity purified polyclonal antibody raised against Cry1Ab protein in rabbits. Native and biotinylated forms of this antibody served as capture antibody and detection antibody for the ELISA, respectively. Streptavidin-horseradish peroxidase conjugate and TMB substrate provided the means for enzymatic colour development.The immunoassay allowed Cry1Ab protein determination in bovine blood plasma in an analytical range of 0.4-100 ng mL−1 with a decision limit (CCα) of 1.5 ng mL−1 and detection capability (CCβ) of 2.3 ng mL−1. Recoveries ranged from 89 to 106% (mean value of 98%) in spiked plasma.In total, 20 plasma samples from cows (n = 7) fed non-transgenic maize and 24 samples from cows (n = 8) fed transgenic maize (collected before and, after 1 and 2 months of feeding) were investigated for the presence of the Cry1Ab protein. There was no difference amongst both groups (all the samples were below 1.5 ng mL−1; CCα). No plasma sample was positive for the presence of the Cry1Ab protein at CCα and CCβ of the assay. |
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Keywords: | Cry1Ab toxin Enzyme immunoassay Bovine blood plasma Bt-maize (MON810) |
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