Highly Sensitive Surface Plasmon Resonance Sensor on Nanoscale Bioactive Surfaces for Specific Detection of Tri-Nitro To
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0951-E07-12
Highly Sensitive Surface Plasmon Resonance Sensor on Nanoscale Bioactive Surfaces for Specific Detection of Tri-Nitro Toluene Praveen Singh1,2, Takeshi Onodera3, Kiyoshi Matsumoto4, Norio Miura5, and Kiyoshi Toko6 1 Department of Electronics, Graduate School of Information Science and Electrical Engineering, Kyushu University, 744 Motooka, Nishi-ku, R.No.459,W2, Fukuoka, 819-0395, Japan 2 Biophysics and Electron Microscopy Section, Indian Veterinary Research Institute, Izatnagar, Bareilly, 243-122, India 3 Department of Electronics, Graduate school of Information Science and Electrical Engineeting, Kyushu University, 744 Motooka, Nishi-ku, Fukuoka, 819-0395, Japan 4 Graduate School of Agriculture, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka, 812-8581, Japan 5 Art, Science and Technology Centre for Cooperative Research, Kyushu University, Kasuga-shi, Fukuoka, 816-8580, Japan 6 Department of Electronics, Graduate School of Information Science and Electrical Engineering, Kyushu University, 744 Motooka, Nishi-ku, Fukuoka, 819-0395, Japan ABSTRACT A nano-scale biosensor chip surface was fabricated using dinitro-phenylated keyhole limpet hemocyanin (DNP-KLH) protein conjugate as ligand supported by underlying 11-amino 1undecanethiol hydrochloride(AUT) self-assembled monolayer (SAM) and bis sulfosuccinimidyl suberate (BS3) as crosslinker. Bioactive thin films were fabricated over gold chip via layer-bylayer self- assembly methods. Biomolecular interaction between substrate specific anti-TNT antibody and DNP-KLH conjugate surface was monitored through surface plasmon resonance based optical sensor. The quantitation of tri-nitro toluene(TNT) on this bioactive surface was done using the solution based competitive inhibition assay. The DNP-KLH surface biosensor has shown a detection limit of 0.14 ng/ml (140 ppt) for TNT molecule. The detection limit of surface plasmon resonance (SPR) biosensor was further enhanced by using goat anti mouse antibody to the 0.002 ng/ml for TNT analyte. This TNT specific biosensor holds the promise to be one of most sensitive sensor surface under indirect competitive assay format. A short injection (12 sec) of 10 mM glycine-HCl solution was found adequate for regeneration of DNP-KLH surface for repeated use. INTRODUCTION Highly selective and sensitive detection of TNT and RDX is the major challenge to scientific community for creation of safety in air travel, cargo handling at sea port, containing the environmental pollution of sea water and land, checking the rise of global terrorism and trafficking in explosives. Millions of landmines are buried in the field and are still being laid every year, thus creating very unsafe environment for human society world over. Trinitrotoluene (TNT) is the main component of these products serves as an indicator for existence of these explosives and is a contaminating molecule. Current physical detection methods are less selective, unreliable and have poor accuracy. There are many attempts to develop reliable sensor technologies using var
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