Highly sensitive immunosensing platform for one-step detection of genetically modified crops.

The wide cultivation of genetically modified (GM) insect-resistant crops has raised concerns on the risks to the eco-environment resulting from a release of Cry proteins. Therefore, it is vital to develop a method for the quantification of GM crops. Herein, A highly sensitive immunosensing platform has been developed for both colorimetric and chemiluminescent (CL) detection of Cry 1Ab using dual-functionalized gold nanoparticles (AuNPs) as signal amplification nanoprobes for the first time. In this work, anti-Cry 1Ab monoclonal antibody and horseradish peroxidase (HRP) are simultaneously functionalized on the surface of AuNPs with an exceptionally simple synthesis method. Combined with immunomagnetic separation, this immunosensing platform based on colorimetric method could detect Cry 1Ab in one step in a linear range from 1.0 to 40 ng mL-1 within 1.5 h, with a limit of detection of 0.50 ng mL-1. The sensitivity of fabricated nanoprobes was 15.3 times higher than that using commercial HRP-conjugated antibody. Meanwhile, the fabricated nanoprobes coupled with CL detection was successfully applied for Cry 1Ab detection with a minimum detection concentration of 0.050 ng mL-1 within a linear range of 0.10-20 ng mL-1. The proposed approach was validated with genuine GM crops, and the results showed a good correlation coefficient of 0.9906 compared to those of a commercial ELISA kit. Compared with ELISA, the developed immunosensing platform significantly simplified the assay procedure and shortened the analytical time, thus providing a new platform for the detection of genetically modified crops with high sensitivity, rapidity and simplicity.

A portable electrochemical immunosensor for highly sensitive point-of-care testing of genetically modified crops.

The wide cultivation of genetically modified (GM) crops has raised concerns on the risks to humans and the environment. 5-enolpyruvylshikimate-3-phosphate synthase isolated from Agrobacterium species strain CP4 (CP4-EPSPS) protein is most widely present in these crops. Therefore the measurement of CP4-EPSPS sensitively in a point-of-care testing (POCT) manner for the screening of transgenic plants is demanded. To date the development of quantitative POCT system has not yet been reported. In presented study, an electrochemical immunosensor towards CP4-EPSPS has been fabricated by integrating a portable bioanalytical device with a disposable screen-printed carbon electrode (SPCE) for POCT of GM crops. The dual-functionalized AuNPs were used as nanoprobes and prepared by simultaneously tagging horseradish peroxidase (HRP) and antibody on AuNPs with an exceptionally simple protocol. The sensitivity of the developed nanoprobe-based immunosensor was 62.5-fold higher than that using HRP-labeled antibody. As a result, the proposed immunosensor using SPCE could detect CP4-EPSPS down to 0.050 ng mL-1 with the linear range of 0.10-10 ng mL-1 within 65 min. In addition, the developed method has been validated with genuine GM crops and the results show a good correlation coefficient of 0.9909 compared with those of a commercial ELISA kit. Therefore, this portable electrochemical immunosensor is suitable for rapid and sensitive detection and provides a convenient and reliable platform for POCT assay.

Sensitive and Facile Electrochemiluminescent Immunoassay for Detecting Genetically Modified Rapeseed Based on Novel Carbon Nanoparticles.

A highly sensitive electrochemiluminescent (ECL) immunoassay targeting PAT/ bar protein was facilely developed for genetically modified (GM) rapeseed detection using carbon nanoparticles (CNPs) originally prepared from printer toner. In this work, CNPs linked with antibody for PAT/ bar protein were used to modify a working electrode. After an immunoreaction between the PAT/ bar protein and its antibody, the immunocomplex formed on the electrode receptor region resulted in an inhibition of electron transfer between the electrode surface and the ECL substance, thus led to a decrease of ECL response. Under the optimal conditions, the ECL responses linearly decreased as the increase of the PAT/ bar protein concentration and the GM rapeseed RF3 content in the ranges of 0.10-10 ng/mL and 0.050-1.0%, with the limits of detection of 0.050 ng/mL and 0.020% (S/N = 3). These results open a facile, sensitive, and rapid approach for the safety control of agricultural GM rape.

An ultrasensitive label-free electrochemiluminescent immunosensor for measuring Cry1Ab level and genetically modified crops content.

The development of genetically modified (GM) insect-resistant crops has aroused great public concern about the risks on the eco-environment resulting from a release of toxic Cry proteins (such as Cry1Ab) to the soil. Therefore, it is of crucial importance to measure the Cry proteins level and the GM crops content. Here, we have tested for the first time a method that uses novel carbon nanospheres (CNPs) label-free electrochemiluminescent (ECL) immunosensor for the ultrasensitive quantification of Cry1Ab and GM crops. In this work, novel CNPs were prepared from printer toner with a very facile approach, and linked with anti-Cry1Ab antibodies to modify a golden working electrode. The immunoreaction between Cry1Ab and its antibody formed an immunocomplex on the bioreceptor region of the sensor, which inhibited electron transfer between the electrode surface and the ECL substance, leading to a decrease of ECL response. Under the optimal conditions, the fabricated label-free ECL immunosensor determined Cry1Ab down to 3.0pgmL-1 within a linear range of 0.010-1.0ngmL-1, showing significant improvement of sensitivity than that of most previous reports. Meanwhile, the proposed method was successfully applied for GM rice BT63 and GM maize MON810 detections down to 0.010% and 0.020%, respectively. Due to its outstanding advantages such as high sensitivity, ideal selectivity, simple fabrication, rapid detection, and low cost, the developed method can be considered as a powerful and pioneering tool for GM crops detection. Its use can also be extended to other toxin protein sensing in foods.