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Transcription-Type Protein Imprinted Polymers for SPR Sensing Prepared Using Target-immobilized Stamps based on Submicrometer-Sized Particles via Biotin-Avidin Linkage

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Yoshizawa S., Kuwata T., Takano E., Kitayama Y., Takeuchi T.
Molecular Imprinting
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2014
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vol. 2
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issue 1
EN
Transcribed molecularly imprinted polymers (MIPs), prepared by using the biotinylated submicrometersized particles-immobilized stamp on which the biotinconjugated protein was hierarchically immobilized via avidin, were used successfully in the recognition of cytochrome c (Cyt) as a model protein. The transcribed MIP for Cyt was prepared on the gold-coated surface plasmon resonance (SPR) sensor chip, and the binding behavior of Cyt, myoglobin, ribonuclease A, lysozyme, and avidin was evaluated to confirm the selectivity for Cyt. The imprint effect of the transcribed MIP was revealed by comparing the MIP and the corresponding non-imprinted polymer prepared using the stamp without the Cyt immobilization.
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A close analysis of metal-enhanced fluorescence of tryptophan induced by silver nanoparticles: wavelength emission dependence

100%
Caires A., Costa L., Fernandes J.
Open Chemistry
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2013
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vol. 11
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issue 1
111-115
EN
In the last few years, silver nanoparticles have been proposed as a promising alternative for the label-free detection of proteins via metal-enhanced fluorescence. Generally, the aromatic amino acid tryptophan is most frequently used in this type of studies, because the intrinsic fluorescence of proteins is usually dominated by tryptophan emissions. In the present study, we evaluated the fluorescence behavior of tryptophan in the presence of a silver colloid with nanoparticles of 100 nm in diameter. The results showed that a nanoparticles concentration of 32 mg L−1 induced maximum fluorescence enhancement. However, the metal-enhanced fluorescence was dependent on the emission wavelength of tryptophan, and this phenomenon was closely related to the metal surface reabsorption process (inner filter effect), suggesting that the plasmon resonance reabsorption effect should be taken into account in analyses involving protein studies by metal-enhanced fluorescence.
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