研究目的
To develop a simple and effective method for the visual detection of denatured proteins using the plasmonic behavior of gold nanoparticles.
研究成果
The research successfully demonstrated a proof of concept for visually detecting denatured ferritin using gold nanoparticles and APTES, offering a fast, low-cost, and user-friendly method for protein degradation detection. This method has potential applications in ensuring the stability of pharmaceutical drug products.
研究不足
The study focuses on ferritin from horse spleen and may not be directly applicable to other proteins without further studies. The visual detection system's sensitivity and specificity for different proteins and under various conditions need further exploration.
1:Experimental Design and Method Selection:
The study utilized the plasmonic properties of gold nanoparticles for visual biosensing of denatured proteins.
2:Sample Selection and Data Sources:
Ferritin type-I from horse spleen was used as the target analyte.
3:List of Experimental Equipment and Materials:
Gold chloride trihydrate, trisodium citrate, APTES, ferritin, and deionized water were used.
4:Experimental Procedures and Operational Workflow:
Gold nanoparticles were synthesized and mixed with ferritin and APTES. The solutions were characterized using UV/Vis/NIR spectrophotometry, dynamic light scattering, Raman spectroscopy, and circular dichroism.
5:Data Analysis Methods:
The absorbance spectra, size distribution, and Raman spectra were analyzed to quantify protein degradation.
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JEOL JEM-2100F high resolution TEM
JEM-2100F
JEOL
Used for transmission electron microscopy imaging of gold nanoparticles and APTES samples.
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Gold chloride trihydrate
HAuCl4?3H2O
Sigma-Aldrich
Used as a precursor for gold nanoparticle synthesis.
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3-aminopropyl triethoxysilane
APTES
Sigma-Aldrich
Used to induce aggregation of gold nanoparticles for protein detection.
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LAMBDA 950 UV/Vis/NIR Spectrophotometer
LAMBDA 950
PerkinElmer
Used to obtain the absorbance spectra of the solutions.
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JASCO J-815 CD Spectrophotometer
J-815
JASCO
Used to obtain circular dichroism spectra of ferritin.
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Trisodium citrate reagent
Alfa Aesar
Used in the synthesis of gold nanoparticles.
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Ferritin, type-I from horse spleen
Sigma-Aldrich
Used as the target analyte for denaturation studies.
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Nanotrac Wave
Microtrac
Used for dynamic light scattering to determine the size and size distribution of ferritin.
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EZRaman-I series high sensitivity portable Raman analyzer
EZRaman-I
Enwave Optronics
Used for Raman spectroscopy to characterize ferritin.
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