研究目的
Developing a simple and efficient enrichment and detection method for metallothionein (MT) to address the time-consuming and complicated traditional processes.
研究成果
The developed method combining Fe3O4@Au core-shell nanoparticles with mass spectrometry technology offers a simple, fast, and sensitive approach for the enrichment and identification of MT, with potential applications in proteomics and related fields.
研究不足
The method's effectiveness is dependent on the concentration of MT, with a detection limit of 10 fg mL?1. The saturation of nanoparticle surfaces may limit enrichment efficiency at higher MT concentrations.
1:Experimental Design and Method Selection
Preparation of core-shell nanoparticles (Fe3O4@Au NPs) for the enrichment of MT through Au-S interaction and identification by MALDI-TOF/MS.
2:Sample Selection and Data Sources
Standard rabbit liver MT and bovine serum albumin (BSA) were used to prepare protein mixture solutions.
3:List of Experimental Equipment and Materials
Nicolet iS5 Infrared Spectrometer, JEM-2010 Transmission Electron Microscope, RW20DZM.n Electronic Constant Speed Stirrer, UV-2540 UV-visible Spectrophotometer, Bruker Daltonics MALDI-TOF/MS, Zetasizer Nano-ZS90 potentiometer, X-ray diffractor.
4:Experimental Procedures and Operational Workflow
Fe3O4 nanoparticles were synthesized and coated with gold to form core-shell nanoparticles. These nanoparticles were then used to enrich MT from protein mixtures, followed by MALDI-TOF/MS analysis.
5:Data Analysis Methods
The enriched proteins were analyzed by MALDI-TOF/MS to identify MT, with the detection limit and specificity evaluated.
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Nicolet iS5 Infrared Spectrometer
iS5
Thermo Fisher
Used for infrared spectroscopy analysis.
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UV-2540 UV-visible Spectrophotometer
UV-2540
Shimadzu Corporation
Used for UV-visible absorption spectroscopy analysis.
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Bruker Daltonics MALDI-TOF/MS
MALDI-TOF/MS
Bruker Daltonics
Used for mass spectrometry identification of proteins.
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Zetasizer Nano-ZS90 potentiometer
Nano-ZS90
Malvern
Used for Zeta potential analysis.
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X-ray diffractor
D2 Phaser
Bruker
Used for X-ray diffraction analysis.
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JEM-2010 Transmission Electron Microscope
JEM-2010
Japan Electronics
Used for transmission electron microscopy analysis.
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RW20DZM.n Electronic Constant Speed Stirrer
RW20DZM.n
IKA
Used for stirring solutions at a constant speed.
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