研究目的
Investigating the use of gold nanoflowers (AuNFs) as probes for in-situ monitoring of alkaline phosphatase (ALP) activity using localized surface plasmon resonance (LSPR) technique.
研究成果
The study successfully demonstrated the use of AuNFs as LSPR probes for ultrasensitive detection of ALP activity in vitro and in living cells. The method offers a new avenue for cell studies and potential applications in discovering ALP-based drugs.
研究不足
The study is limited by the specificity of the AuNFs probe to ALP activity and the potential interference from other cellular components. The technique's sensitivity and resolution in complex biological samples need further optimization.
1:Experimental Design and Method Selection:
The study involved the synthesis of AuNFs through oxidative disruption of polydopamine-coated Au nanoparticles and subsequent growth of Au nanopetals. The LSPR technique was employed for monitoring ALP activity.
2:Sample Selection and Data Sources:
HepG2 and HEK 293 cells were used as model cell lines for ALP activity monitoring.
3:List of Experimental Equipment and Materials:
Gold nanoparticles, polydopamine, HAuCl4, polyvinylpyrrolidone (PVP), hydroxyl amine, AgNO3, L-ascorbic acid 2-phosphate sesquimagnesium salt hydrate (AAP), and ALP were used.
4:Experimental Procedures and Operational Workflow:
AuNFs were synthesized and characterized. ALP activity was monitored by the deposition of Ag on AuNFs surface, which was catalyzed by ALP. The LSPR response and scattering color changes were measured.
5:Data Analysis Methods:
The LSPR peak shifts and scattering color changes were analyzed to determine ALP activity.
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Gold nanoparticles
Au13
Used as seeds for the synthesis of larger gold nanoparticles (Au50).
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Gold nanoparticles
Au50
Core particles for the synthesis of gold nanoflowers (AuNFs).
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Polydopamine-coated Au50
PDA/Au50
Intermediate product in the synthesis of AuNFs, providing a surface for nanopetal growth.
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Gold nanoflowers
AuNFs
Probes for in-situ monitoring of ALP activity using LSPR technique.
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Silver nitrate
AgNO3
Source of Ag+ for deposition on AuNFs surface, catalyzed by ALP activity.
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L-ascorbic acid 2-phosphate sesquimagnesium salt hydrate
AAP
Substrate for ALP, hydrolyzed to produce L-ascorbic acid (AA) which reduces Ag+ to Ag.
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Alkaline phosphatase
ALP
Enzyme whose activity is monitored via Ag deposition on AuNFs.
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