Validation of an Immunoassay for Detection and Quantitation of a Genetically Modified Soybean in Food and Food Fractions Using Reference Materials: Interlaboratory Study

Lipp, Markus; Anklam, Elke; Stave, James W.; Bahrs-Windsberger, J; Crespo, M.T.; Benvenuto, E; Berben, Gilbert; Brignon, P.; Busch, Ulrich; Cordes, C; Eklund, Eivor; Jaziri, Mondher El; Engel, Karl‐Heinz; Eriksen, Folmer Damsted; Eriksson, Susann; Felinger, Attila; Fogher, Corrado; Folch, I; Garrett, Steve; Hischenhuber, Claudia; Hörtner, H; Jany, K.-D.; Krska, R.; Kruse, Linda; Kuiper, H.A.; Lobre, C; Miraglia, Marina; Mhller, W; Costa, João; Oliveira, Luciana M.; Patel, Pradip D.; Pietsch, Klaus; Pöpping, Bert; Remler, P; Rentsch, Jürg; Schulze, Marcel; Sutton, Mark A.; Duijn, G. Van; Kan, F van; Wurz, A.

doi:10.1093/jaoac/83.4.919

Cited by 78 publications

(24 citation statements)

References 2 publications

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“…Other methods are based on the detection of proteins specifically expressed in GM crops. Direct double‐antibody (preferred) and indirect triple‐antibody sandwich ELISA formats are most frequently employed to detect and measure the levels of novel proteins produced in GM plant varieties (Allen, 1990; Lipp et al. , 2000).…”

Section: Discussionmentioning

confidence: 99%

A simple and reliable assay for detecting specific nucleotide sequences in plants using optical thin‐film biosensor chips

Bai

Zhong

et al. 2006

The Plant Journal

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SummaryHere we report the adaptation and optimization of an efficient, accurate and inexpensive assay that employs custom-designed silicon-based optical thin-film biosensor chips to detect unique transgenes in genetically modified (GM) crops and SNP markers in model plant genomes. Briefly, aldehyde-attached sequence-specific single-stranded oligonucleotide probes are arrayed and covalently attached to a hydrazine-derivatized biosensor chip surface. Unique DNA sequences (or genes) are detected by hybridizing biotinylated PCR amplicons of the DNA sequences to probes on the chip surface. In the SNP assay, target sequences (PCR amplicons) are hybridized in the presence of a mixture of biotinylated detector probes and a thermostable DNA ligase. Only perfect matches between the probe and target sequences, but not those with even a single nucleotide mismatch, can be covalently fixed on the chip surface. In both cases, the presence of specific target sequences is signified by a color change on the chip surface (gold to blue/purple) after brief incubation with an anti-biotin IgG horseradish peroxidase (HRP) to generate a precipitable product from an HRP substrate. Highly sensitive and accurate identification of PCR targets can be completed within 30 min. This assay is extremely robust, exhibits high sensitivity and specificity, and is flexible from low to high throughput and very economical. This technology can be customized for any nucleotide sequence-based identification assay and widely applied in crop breeding, trait mapping, and other work requiring positive detection of specific nucleotide sequences.

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Section: Discussionmentioning

confidence: 99%

A simple and reliable assay for detecting specific nucleotide sequences in plants using optical thin‐film biosensor chips

Bai

Zhong

et al. 2006

The Plant Journal

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“…These methods mainly based on detection of novel DNA or protein present in the product. DNA-based methods widely used both qualitatively and quantitatively for the detection of GMOs product in transgenic raw or unprocessed soy products (Meyer et al, 1996;Lipp et al, 2000;Lipp et al, 2001;Taverniers et al, 2001). Qualitative testing is used to identify GM or non GM material or to distinguish certified or noncertified material.…”

Section: Introductionmentioning

confidence: 99%

The effect of processing factors on detection of genetically modified soy in flour by ELISA assay

Arun¹,

Eker²,

Muratoğlu³

2018

Journal of Istanbul Veterinary Sciences

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Genetic modification (GM) techniques have been an important research area of food and feed industry since the 19th century. There is a strong consumer concern over genetically modified organisms (GMOs) because of their potential risks on health and environment. For this purpose, various countries including Turkey have released labelling regulations for products derived from GMOs. These legal enforcements brought the necessity for reliable detection methods. The aim of our study was to evaluate the effect of processing factors on the detection possibility of GMOs by using a commercial Enzyme Linked Immunosorbent (ELISA) assay. For this, flour mixtures containing 0.5%, 1%, 5%, 10%, 100% were prepared by mixing the appropriate amount of RUR-GM and non GM standard soy flour and main processing techniques most used in the food industry (baking, autoclaving and freezing) were applied. According to our results, the detection of GMOs was possible at all concentrations of autoclaved and frozen samples. In dry heated samples, GMOs could not be detected containing below 5% GMOs. ELISA method cannot be recommended as a reference method for evaluation of the compliance with the regulations, but it can serve as a practical alternative to be used as an online monitoring tool in production lines for raw and mildly processed foods.

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“…Both monoclonal antibodies with high specificity and polyclonal antibodies with high sensitivity can be used depending on the amounts needed and the specificity of the detection system. The immunoassay methods were hardly reported for detection and quantitation of proteins expressed by most biotechnology derived crops with the exception of the Bt Cry1Ab protein in MON810 corn (Stave 2002), the CP4-EPSPS protein in Roundup Ready soybeans (Rogan et al 1999;Lipp et al 2000), the neomycin phosphotransferase II enzyme (npt II) in some plant species (McKenzie et al 2000) and the pat protein in Bt maizes and MS rapes (Xu et al 2005).…”

Section: Introductionmentioning

confidence: 99%

Application of Stepwise Ammonium Sulfate Precipitation as Cleanup Tool for an Enzyme-Linked Immunosorbent Assay of Glyphosate Oxidoreductase in Genetically Modified Rape of Gt73

Xu¹,

Huang²,

Liang³

et al. 2009

Journal of Food Biochemistry

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The method of enzyme‐linked immunosorbent assay after stepwise ammonium sulfate (AS) purification (AS‐ELISA) was developed and used to detect genetically modified (GM) rape of GT73 containing glyphosate oxidoreductase (Gox). Gox protein encoded by the Gox gene from Achromobacter sp. was highly expressed as inclusion bodies in Escherichia coli BL21 (DE3) and purified to homogeneity by Ni2+affinity chromatography. A simple and efficient extraction and purification procedure of Gox protein from the seeds and leaves of GM rape was developed by means of stepwise AS precipitation. Purified polyclonal antibodies against Gox was produced and enzyme‐linked immunosorbent assay (ELISA) procedures were established further on to measure the Gox protein. AS‐ELISA allowed 5% GMOs to be detected in the seeds of GT73 and 0.5% GMOs to be detected in the leaves of GT73 rape, which makes this method an acceptable method to access Gox protein in GM rape of GT73. PRACTICAL APPLICATIONS Many GMOs containing Gox gene have been approved worldwide such as GT73 rape, 1,445 cotton and Mon832 maize. Protein based methods were more important than DNA based methods, because protein performs a specific and concrete function and is closely interconnected with crop traits. AS‐ELISA method can be used in the screening of GM plant, Gox protein expression assay and quantitative detection for GMO labeling. AS‐ELISA Gox detecting method was established in this paper and was being evaluated of Inter‐laboratory Comparison in some of Chinese GMO detection and assessment centers. With the knowledge of ELISA, ELISA method will be the national standards and international and will be a beneficial supplement for the DNA based GMO detecting methods.

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Validation of an Immunoassay for Detection and Quantitation of a Genetically Modified Soybean in Food and Food Fractions Using Reference Materials: Interlaboratory Study

Cited by 78 publications

References 2 publications

A simple and reliable assay for detecting specific nucleotide sequences in plants using optical thin‐film biosensor chips

A simple and reliable assay for detecting specific nucleotide sequences in plants using optical thin‐film biosensor chips

The effect of processing factors on detection of genetically modified soy in flour by ELISA assay

Application of Stepwise Ammonium Sulfate Precipitation as Cleanup Tool for an Enzyme-Linked Immunosorbent Assay of Glyphosate Oxidoreductase in Genetically Modified Rape of Gt73

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