TITLE

Development and validation of a sensitive and fast chemiluminescent enzyme immunoassay for the detection of genetically modified maize

AUTHOR(S)
Roda, A.; Mirasoli, M.; Guardigli, M.; Michelini, E.; Simoni, P.; Magliulo, M.
PUB. DATE
March 2006
SOURCE
Analytical & Bioanalytical Chemistry;Mar2006, Vol. 384 Issue 6, p1269
SOURCE TYPE
Academic Journal
DOC. TYPE
Article
ABSTRACT
Proteins from the Cry 1 family, in particular Cry 1Ab, are commonly expressed in genetically modified Bt maize in order to control chewing insect pests. A sensitive chemiluminescent sandwich enzyme immunoassay for the detection of Cry1Ab protein from genetically modified Bt maize has been developed and validated. A Cry1Ab protein-specific antibody was immobilized on 96- or 384-well microtiter plates in order to capture the Cry1Ab toxin in the sample; the bound toxin was then detected by employing a second anti-Cry1Ab antibody and a horseradish peroxidase-labeled anti-antibody, followed by measurement of the enzyme activity with an enhanced chemiluminescent system. The chemiluminescent assay fulfilled all the requirements of accuracy and precision and exhibited limits of detection of a few pg mL-1 Cry1Ab (3 or 5 pg mL-1, depending on the assay format), which are significantly lower than that achievable using conventional colorimetric detection of peroxidase activity and also represent an improvement compared to previously developed Cry1Ab immunoassays. High-throughput analysis can be performed using the 384-well microtiter plate format immunoassay, which also allows one to reduce the consumption of samples and reagents. Validation of the assay, performed by analyzing certified reference materials, proved that the immunoassay is able to detect the presence of the Cry1Ab protein in certified reference samples containing as low as 0.1% of MON 810 genetically modified Bt maize. This value is below the threshold requiring mandatory labeling of foods containing genetically modified material according to the actual EU regulation.
ACCESSION #
21909060

 

Related Articles

  • Detection of Cry1Ab toxin in the leaves of MON 810 transgenic maize. Székács, András; Lauber, Éva; Takács, Eszter; Darvas, Béla // Analytical & Bioanalytical Chemistry;Mar2010, Vol. 396 Issue 6, p2203 

    The distribution of Cry1Ab toxin was detected in the leaves of genetically modified maize of genetic event MON 810 by enzyme-linked immunosorbent assay. Cry1Ab toxin contents in the leaves at reproductive (milk, R3) phenological stage were measured to be between 3,878 and 11,148 ng Cry1Ab...

  • Synthesis and expression of recombinant interferon alpha-5 gene in tobacco chloroplasts, a non-edible plant. Khan, Muhammad; Nurjis, Farwa // Molecular Biology Reports;Apr2012, Vol. 39 Issue 4, p4391 

    The production of interferon alpha from microbial to mammalian expression system, have certain precincts in terms of cost, scalability, safety and authenticity. Modern biotechnology exploits transgenic crops to get large quantities of complex proteins in a cost-effective way. In order to...

  • High-Throughput Screening for mTORC1/mTORC2 Kinase Inhibitors Using a Chemiluminescence-Based ELISA Assay. Shripad V. Bhagwat; Jennifer Kahler; Yan Yao; Paul Maresca; Maureen Brooks; Andy Crew; Michael Boisclair; Jonathan A. Pachter // Assay & Drug Development Technologies;Oct2009, Vol. 7 Issue 5, p471 

    A high-throughput chemiluminescence and ELISA-based biochemical assay to identify mTORC1/mTORC2 kinase inhibitors is described. These mTOR complexes were isolated from HeLa whole cell lysate using mTOR antibodies and in-well immunoprecipitation. The integrity and purity of the mTORC1 and mTORC2...

  • Serum HER2 levels determined by two methods in patients with metastatic breast cancer. Hayashi, Naoki; Nakamura, Seigo; Tokuda, Yasuharu; Yagata, Hiroshi; Yoshida, Atsushi; Ota, Hidekazu; Hortobagyi, Gabriel; Cristofanilli, Massimo; Ueno, Naoto // International Journal of Clinical Oncology;Feb2012, Vol. 17 Issue 1, p55 

    Background: The role and the optimal measurement method of serum HER2 levels are not defined in patients with metastatic breast cancer (MBC). We prospectively assessed the prognostic value of serum HER2 levels in MBC using two methods, enzyme immunoassay (EIA) and chemiluminescence immunoassay...

  • Accumulation of functional recombinant human coagulation factor IX in transgenic soybean seeds. Cunha, Nicolau B.; Murad, André M.; Ramos, Gustavo L.; Maranhão, Andréia Q.; Brígido, Marcelo M.; Araújo, Ana Cláudia G.; Lacorte, Cristiano; Aragão, Francisco J. L.; Covas, Dimas T.; Fontes, Aparecida M.; Souza, Gustavo H. M. F.; Vianna, Giovanni R.; Rech, Elíbio L. // Transgenic Research;Aug2011, Vol. 20 Issue 4, p841 

    The seed-based production of recombinant proteins is an efficient strategy to achieve the accumulation, correct folding, and increased stability of these recombinant proteins. Among potential plant molecular farming systems, soybean [ Glycine max (L.) Merrill] is a viable option for the...

  • Comparative proteomics of Bt-transgenic and non-transgenic cotton leaves. Limin Wang; Xuchu Wang; Xiang Jin; Ruizong Jia; Qixing Huang; Yanhua Tan; Anping Guo // Proteome Science;9/4/2015, Vol. 13 Issue 1, p1 

    Background: As the rapid growth of the commercialized acreage in genetically modified (GM) crops, the unintended effects of GM crops' biosafety assessment have been given much attention. To investigate whether transgenic events cause unintended effects, comparative proteomics of cotton leaves...

  • P19-dependent and P19-independent reversion of F1-V gene silencing in tomato. M. Alvarez; Heidi Pinyerd; Emel Topal; Guy Cardineau // Plant Molecular Biology;Sep2008, Vol. 68 Issue 1/2, p61 

    Abstract  As a part of a project to develop a plant-made plague vaccine, we expressed the Yersinia pestis F1-V antigen fusion protein in tomato. We discovered that in some of these plants the expression of the f1-v gene was undetectable in leaves and fruit by ELISA, even though they had...

  • Molecular analysis of transgenic melon plants showing virus resistance conferred by direct repeat of movement gene of Cucumber green mottle mosaic virus. Emran, Ali; Tabei, Yutaka; Kobayashi, Kappei; Yamaoka, Naoto; Nishiguchi, Masamichi // Plant Cell Reports;Aug2012, Vol. 31 Issue 8, p1371 

    Cucumber green mottle mosaic virus (CGMMV) is a major limiting factor in the production of melon plants worldwide. For effective control of this virus using the transgenic approach, the direct repeat of the movement protein gene of CGMMV was used for transforming melon plants by Agrobacterium...

  • Genetically modified soybeans: false-positive detection in fermented natural soybean (tempe). Prakoso, Budi; Nitisinprasert, Sunee; Stevens, Willem F. // Biotechnology Letters;Sep2003, Vol. 25 Issue 18, p1485 

    Tempe was prepared using mixtures of natural soybean and genetically modified Roundup Ready (RUR) soybean fermented with natural Rhizopus sp. The amount of RUR soybean was quantified using an ELISA plate test. The RUR signal decreased during fermentation. In the control experiments on...

Share

Read the Article

Courtesy of THE LIBRARY OF VIRGINIA

Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics