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Test kits for traceability of genetically modified crops expressing Cry 1Ab protein

Identification

Key words Traceability, Bacillus Thuringiensis, Cry 1Ab, immunodiagnostic, transgenic, GMO, maize
Latest version 2012/12/07
Completed by INRA - IATE

How does it work?

Primary objective Detection of the presence of transgenic maize (through new immunodetection tools for of the Cry 1Ab protein of Bacillus thuringiensis expressed in some transgenic maize, e.g. Monsanto 810).
Working principle The Cry 1Ab protein is produced only by transgenic varieties of maize. Consequently, identifying this protein is a 100% sure sign for the presence of genetically modified maize in a food (seeds, leaves, flour). The detection method uses monoclonal antibodies with different specificities directed against the purified Cry 1Ab protein from the Bacillus thuringiensis. 5 monoclonal antibodies (Mab#40, 73, 95, 115, 120) having different specificities for the native or denatured Cry 1Ab protein have been purified. They can be used according to various combinations to develop the most appropriate immunological detection test(s).

So far, one sandwich-ELISA test kit has been designed with two selected antibodies; this ELISA test is highly specific and sensitive to the native Cry1Ab protein. As a result, the test kit allows to detect the presence of the Cry 1Ab protein in a sample with a sensitivity of 0.1% (e.g. 1 GM seed in a 1000 seed lot).

Images
Additional effects Other test kits (e.g. ELISA, LFDs, etc.) can be designed with more monoclonal antibodies, in order to identify any vegetable material coming from genetically modified crops that express the Cry 1Ab protein. The manufacturer has the possibility to implement various immunodetection tests based on these monoclonal antibodies: ELISA, FLDs, etc.

Monoclonal antibodies directed against the Cry 1Ab protein from the B. thuringiensis were produced: 5 of them were selected, having different specificities for the native or denatured Cry 1Ab protein.

Important process parameters - preparation of the sample (grinding, milling)

- composition of the extraction buffer

- incubation

- preservation of extraction product

Important product parameters Product: detection tests can be performed on maize seeds, leaves, flour

biological materials: monoclonal antibodies

What can it be used for?

Products Any product can be tested, but it is worthless testing a product you don’t suspect as potentially containing GMO.

Maize: seeds, leaves, flour To be tested: processed flour-based products (e.g.: bread)

Operations Other: quality control, authentication
Solutions for short comings GMO control

What can it NOT be used for?

Products None (see “product” section on previous tab)
Operations The test has to be assessed for cooked food.
Other limitations Only the Cry 1Ab protein is identified; it can therefore only be applied to samples where this specific protein is suspected. If another genetically modified crop, not expressing the Cry 1Ab protein, has contaminated the lot, this test is useless.
Risks or hazards No, standard immunodiagnostics

Implementation

Maturity The 5 monoclonal antibodies are owned and available for transfer through Inra Transfert. No license has been exploited so far (December 2012).
Modularity /Implementation This technology should improve the existing tests (MON 810 detection tests already exists) by Detection and quantification of the Cry 1Ab protein of Bacillus thuringiensis expressed in transgenic plants.

It has been determined that this ELISA is highly specific and sensitive to the native Cry1Ab, with a limit of quantification below 1 seed of Monsanto 810 GM maize admixed in more than 1,000 non-GM maize seeds.

The development of new kits could allow an easy, rapid, specific and very sensitive detection (about 0.1%) of the Cry 1Ab protein, and traceability of transgenic crops expressing this Bt toxin, whatever the technology implemented and the sample (ELISA, LFDs, etc.).

Consumer aspects Interest for non-GMO food/feed
Legal aspects The launch on the market of GMOs is strongly regulated in Europe. A list of the commercially available or currently field-tested GMOs can be obtained with the responsible authorities.

Please check local legislation.

Environmental aspects The environmental debate about GMOs is more focused on earlier stages of the food production chain (field)

Further Information

Institutes CEA - iBiTec - S
Companies INRA Transfert
References European Integrated Project Co-Extra ("GM and Not-GM food supply chains: to their

CO-EXistence and TRAceability"; http://www.coextra.eu/)

- preparation of the sample (grinding, milling)

- composition of the extraction buffer

- incubation

- preservation of extraction productwarning.png"- preparation of the sample (grinding, milling)

- composition of the extraction buffer

- incubation

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Created by Hte inra on 7 December 2012, at 19:36