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Dhesingh Ravi Shankaran Toshikazu Kawaguchi Sook Jin Kim Kiyoshi Matsumoto Kiyoshi Toko Norio Miura 《International journal of environmental analytical chemistry》2013,93(10-11):771-781
Recent concern on international terrorism and weapons of mass destruction demands the development of novel analytical methods for identification and quantification of explosive molecules. In this article, we describe the development of high-performance immunosensors for detection of 2,4,6-trinitrotoluene (TNT), a prime component of the landmines and bombs used by terrorist and military forces. The immunosensors were constructed by physical adsorption and self-assembly methods, and their binding interactions with a monoclonal anti-TNT antibody were evaluated for TNT detection using the surface plasmon resonance technique. A home-made 2,4,6-trinitrophenyl-keyhole limpet hemocyanine conjugate was used for physical adsorption. A poly(ethylene glycol) hydrazine hydrochloride thiolate was used in the construction of self-assembled monolayer surface and was immobilized with trinitrophenyl-β-alanine by the amide coupling method. The immunosensors were highly selective, regenerable, rapid, and exhibited remarkable sensitivity down to the parts-per-trillion level for TNT by the indirect competitive inhibition principle. 相似文献
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Dhesingh Ravi Shankaran 《Analytica chimica acta》2003,478(2):321-327
A new silver dispersed composite electrode made of sol-gel derived ceramic-graphite has been demonstrated for the determination of dopamine. Silver has been immobilized within the porous and rigid silicate network in the composite by the specific interaction with (3-mercaptopropyl)trimethoxysilane (MPS). The modified composite electrode coated with Nafion exhibited good catalytic activity for the oxidation of dopamine at a reduced potential of 0.35 V with good sensitivity and selectivity. The sensor showed a linear response to dopamine in the concentration range from 6.6×10−6 to 1.2×10−3 M with a correlation coefficient of 0.9987. The electrode surface can be easily renewed by a simple mechanical polishing and has the advantages of good reproducibility, rapid response and remarkable stability. 相似文献
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Shankaran DR Kawaguchi T Kim SJ Matsumoto K Toko K Miura N 《Analytical and bioanalytical chemistry》2006,386(5):1313-1320
Detection of TNT is an important environmental and security concern all over the world. We herein report the performance and
comparison of four immunoassays for rapid and label-free detection of 2,4,6-trinitrotoluene (TNT) based on surface plasmon
resonance (SPR). The immunosensor surface was constructed by immobilization of a home-made 2,4,6-trinitrophenyl–keyhole limpet
hemocyanin (TNPh–KLH) conjugate onto an SPR gold surface by simple physical adsorption within 10 min. The immunoreaction of
the TNPh–KLH conjugate with four different antibodies, namely, monoclonal anti-TNT antibody (M-TNT Ab), monoclonal anti-trinitrophenol
antibody (M-TNP Ab), polyclonal anti-trinitrophenyl antibody (P-TNPh Ab), and polyclonal anti-TNP antibody (P-TNP Ab), was
studied by SPR. The principle of indirect competitive immunoreaction was employed for quantification of TNT. Among the four
antibodies, the P-TNPh Ab prepared by our group showed highest sensitivity with a detection limit of 0.002 ng/mL (2 ppt) TNT.
The lowest detection limits observed with other commercial antibodies were 0.008 ng/mL (8 ppt), 0.25 ng/mL (250 ppt), and
40 ng/mL (ppb) for M-TNT Ab, P-TNP Ab, and M-TNP Ab, respectively, in the similar assay format. The concentration of the conjugate
and the antibodies were optimized for use in the immunoassay. The response time for an immunoreaction was 36 s and a single
immunocycle could be done within 2 min, including the sensor surface regeneration using pepsin solution. In addition to the
quantification of TNT, all immunoassays were evaluated for robustness and cross-reactivity towards several TNT analogs.
相似文献
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Toshikazu Kawaguchi Dhesingh Ravi Shankaran S.J. Kim Kiyoshi Matsumoto Norio Miura 《Talanta》2007,72(2):554-560
We have developed a new immunosensor based on self-assembly chemistry for highly sensitive and label-free detection of 2,4,6-trinitrotoluene (TNT) using surface plasmon resonance (SPR). A monolayer of amine terminated poly(ethylene glycol) hydrazinehydrochloride (PEG-NH2) thiolate was constructed on an activated gold surface and immobilized with trinitrophenyl-β-alanine (TNPh-β-alanine) by amide coupling method. The binding interaction of a monoclonal anti-TNT Ab (M-TNT Ab) with TNPh-β-alanine immobilized thiolate monolayer surface was monitored and evaluated for detection of TNT based on the principle of indirect competitive immunoreaction. Here, the competition between the self-assembled TNT derivative and the TNT in solution for binding with antibody yields in the response signal that is inversely proportional to the concentration of TNT in the linear detection range. With the present immunoassay format, TNT could be detected in the concentration range from 0.008 ng/ml (8 ppt) to 30 ng/ml (30 ppb). The response time for an immunoreaction was 2 min and one immunocycle could be done with in 4 min including surface regeneration. Bound antibodies could be easily eluted from the self-assembled immunosurface at high recoveries (more than 100 cycles) using pepsin solution without any damage to the TNT derivatives immobilized on the surface. The compact self-assembled monolayer was highly stable and prevented the non-specific adsorption of proteins on the surface favoring error free measurement. 相似文献
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