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Synthesis of Functionalized Silica Particles for Label‐free Detection of PTP1B Using FRET
Author(s) -
Durgannavar Trishaladevi,
Ahn Dohee,
Hwang Ji Young,
Yoon SunYoung,
Kang Hyo Jin,
Chung Sang J.
Publication year - 2019
Publication title -
bulletin of the korean chemical society
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.237
H-Index - 59
ISSN - 1229-5949
DOI - 10.1002/bkcs.11888
Subject(s) - förster resonance energy transfer , fluorescence , detection limit , chemistry , acceptor , tryptophan , nanoparticle , lysis , fluorescence microscope , photochemistry , biophysics , nanotechnology , materials science , chromatography , biochemistry , physics , amino acid , quantum mechanics , biology , condensed matter physics
Silica nanoparticles (SiNPs, 30 and 70 nm) and microparticles (SiMPs, 10 μm) were functionalized with a PTP1B specific probe for label‐free fluorescence detection of PTP1B. Intrinsic tryptophan residues of PTP1B serve as the Förster resonance energy transfer (FRET) donor and the functionalized silica particles as the FRET acceptor. When a mixture of the functionalized SiNP and PTP1B is excited at tryptophan excitation maximum (280 nm), significant sensitized fluorescence was detected at 450 nm as a result of FRET. The limit of detection of PTP1B was found to be 29 and 70 nM for the functionalized 30 and 70 nm SiNP, respectively. Using this approach, PTP1B could be detected in E. coli cell lysate in which PTP1B was induced. The functionalized SiNP probe (SiNP/NA) was selective toward PTP1B over BSA, and the nanoparticles are able to penetrate into cells. Furthermore, the binding of PTP1B to the SiMPs could be visualized under a deep UV‐microscope.