NSTI BioNano 2010

Detection of aflatoxin B1 by SPR biosensor using fusion proteins as a linker

S. Ko, A.-R. Kim, C.J. Kim, D.Y. Kwon
Korea Food Research Institute, KR

Keywords: aflatoxin B1, SPR immunosensor, fusion protein

Abstract:

Because of the health risks of aflatoxin B1 (ATB1), it is essential to monitor the level of this mycotoxin in a variety of foods. HPLC and ELISA methods require skilled personnel, expensive equipments, and complex sample preparations for detection of AFB1, so simple, sensitive, and portable devices are urgently required in the fields. One of the main issues in the development of surface plasmon resonance (SPR) immunosensors is efficient immobilization of antibodies. Antibodies have been immobilized on the sensor surfaces by various techniques, but these methods are cumbersome and cause antibodies to be random oriented. In this work, a novel fusion protein was constructed by genetically fusing gold binding polypeptides (GBP) to protein A (ProA), and SPR immunosensor for detection of ATB1 was simply fabricated by using this fusion protein as a crosslinker for effective immobilization of antibodies. Consequently, a low detection limit (10 ng/ml) has been achieved for mycotoxin SPR immunosensor by using GBP-ProA fusion proteins as a crosslinker. These results indicate the GBP-ProA protein could be a valuable crosslinker for simple and oriented immobilization of antibodies onto SPR gold surfaces, and this SPR immunosensor can become a useful analytical tool for rapid and real-time detection of ATB1.
 
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