研究目的
To describe the first preparation of neoglycoprotein functionalized fluorescent gold nanoclusters and demonstrate their utility for specific sensing of plant lectins and in vitro imaging of dendritic cells.
研究成果
The study presents the first synthesis of neoglycoprotein-protected gold nanoclusters and evaluates their physical and optical properties. The utility of G0-OVA-AuNCs for the analysis of carbohydrate-protein interactions and in vitro imaging of dendritic cells was demonstrated.
研究不足
Further experiments are needed to better understand the role of glycan in the uptake of nanoclusters by dendritic cells and to study their bio- and cytocompatibility.
1：Experimental Design and Method Selection:
The synthesis of neoglycoprotein functionalized fluorescent gold nanoclusters (G0-OVA-AuNCs) was optimized based on previous reports. The protein scaffold acts as both reductive and stabilizing reagent.
2：Sample Selection and Data Sources:
Ovalbumin (OVA) was used as a carrier/model antigen, and a biantennary N-glycan G0 as a targeting molecule.
3：List of Experimental Equipment and Materials:
Gold (III) chloride trihydrate, sodium hydroxide, and triethylamine were purchased from Sigma Aldrich. LPS-free ovalbumin was purchased from Hyglos.
4：Experimental Procedures and Operational Workflow:
The synthesis involved the addition of gold tetrachloroauric (III) acid to a protein solution, followed by 1 M aqueous solution of sodium hydroxide. The mixture was then incubated at 100 °C for 6 min in a microwave reactor.
5：Data Analysis Methods:
The formation of OVA-AuNCs was confirmed by UV-Vis spectroscopy, fluorescence emission spectra, TEM, and agarose gel electrophoresis.
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Disuccinimidyl suberate ester
DSS
Thermo Fisher Scientific
Crosslinking reagent
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Slide-Z-lyser TM dialysis cassettes
10 K MWCO
Thermo Fisher Scientific
Dialysis of AuNCs
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Gold (III) chloride trihydrate
Sigma Aldrich
Used in the synthesis of gold nanoclusters
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LPS-free ovalbumin
Hyglos
Used as a carrier/model antigen
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Nunc? 96-Well Polystyrene Black plates
Thermo Scientific
Measurement of fluorescence emission spectra
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Varioskan Flashmicroplate reader
Thermo Scientific
Measurement of fluorescence emission spectra
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JOEL JEM-2100F field emission TEM
JOEL
TEM imaging
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Copper TEM grids
Ted Pella
TEM imaging
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ImageJ
Quantification of the average diameter of the gold nanocluster
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Biotage? Initiator microwave reactor
Biotage
Incubation of the mixture at 100 °C for 6 min
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Zeiss LSM 510 laser scanning confocal microscope
Carl Zeiss
Confocal microscopy
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