Abstract
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.
Highlights
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
AuNPs conjugated to horseradish peroxidase (HRP)-labeled antibodies have been applied in immunoassays[20]
We developed an exceptionally simple and sensitive immunosensing platform targeting Cry 1Ab for the verification of GM crops based on a AuNP-triggered enzyme signal amplification system and immunomagnetic separation strategy
Summary
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. Various analytical methods have been developed for GM crop detection, such as the polymerase chain reaction (PCR) assay[1,2,3,4,5], quartz crystal microbalance biosensors[6], surface plasma resonance biosensors[7,8], electrochemical biosensors[9,10] and electrochemiluminescent biosensors[11] These DNA-based techniques are reliable, accurate and highly sensitive, they require laborious sample pretreatments and expensive instrumentation. We developed an exceptionally simple and sensitive immunosensing platform targeting Cry 1Ab for the verification of GM crops based on a AuNP-triggered enzyme signal amplification system and immunomagnetic separation strategy. In this investigation, both anti-Cry 1Ab monoclonal antibody and HRP were independently combined onto AuNPs. As-prepared dual-functionalized AuNPs were employed as signal amplification probes to enhance detection sensitivity. Owing to the application of HRP as the signal probe, CL as well as colorimetric methods could be adopted to quantitatively detect GM crops with a high sensitivity
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