研究目的
To develop a fluorescent turn-on sensor for the detection of lectins and Escherichia coli using self-assembled glycobisacrylamide-stabilized gold nanoparticles.
研究成果
The self-assembled glucose based bisacrylamide (Glc-bis) can be utilized for synthesizing stable AuNPs having shelf life of more than 6 months at 25 °C. The fluorescence quenched system, Glc-bis@AuNPs, derived by the quenching of intrinsic emission of Glc-bis aggregates by AuNPs, could be exploited in the development of a fluorescent turn-on sensor for the detection of lectins and Escherichia coli. The turn-on of the quenched emission of Glc-bis@AuNPs upon interaction with Con A was studied by steady state as well as fluorescence lifetime measurements. The binding constant of Glc-bis@AuNPs with Con A was 9.8 × 104 M-1 and the LOD value was found to be 1.6 nM. Glc-bis@AuNPs can detect Con A with high sensitivity and remarkable selectivity in the presence of other proteins and lectins.
研究不足
The study is preliminary and requires further validation with real samples of E.coli/lectins to ascertain the applicability of this method. Converting this method of detection into a strip/solid state based sensor for protein/bacterial detection is still in progress.
1:Experimental Design and Method Selection:
Synthesis of Glc-bis stabilized AuNPs (Glc-bis@AuNPs) by in-situ reduction and stabilization using a D-glucose based bisacrylamide solution.
2:Sample Selection and Data Sources:
Use of Concanavalin A (Con A) and Escherichia coli (E.coli) as model proteins and bacteria, respectively.
3:List of Experimental Equipment and Materials:
TEM, UV-vis spectrophotometer, fluorimeter, FTIR spectrophotometer, zeta potential measurements, dynamic light scattering measurements, fluorescent microscope.
4:Experimental Procedures and Operational Workflow:
Preparation of Glc-bis@AuNPs, interaction with Con A and E.coli, measurement of fluorescence and absorption spectra.
5:Data Analysis Methods:
Calculation of binding constants, limit of detection (LOD), and analysis of fluorescence lifetime measurements.
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TEM
Libra-120
Zeiss-Carl
Transmission Electron Microscope
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FTIR spectrophotometer
Affinity-1
Shimadzu
FTIR measurements
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Fluorescent microscope
BX53F
Olympus
fluorescent imaging
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Jasco FP 8500 fluorimeter
FP 8500
Jasco
steady state fluorescence measurements
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Jasco V-650 spectrophotometer
V-650
Jasco
UV-visible spectroscopy
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FEG-TEM
Technai G2, F30
FEI
Field Emission Gun - Transmission Electron Microscope
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Zetasizer
Malvern
zeta potential measurements
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VASCO γ-particle size analyzer
dynamic light scattering measurements
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Time correlated single photon counting (TCSPC) spectrometer
IBH (UK)
time resolved studies
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