研究目的
To develop a robust covalent coupling scheme for the development of FRET aptasensor based on amino-silane modified graphene oxide to overcome the limitations of physisorbed aptasensors such as poor signal reversibility and reproducibility, and nonspecific probe displacement.
研究成果
The study successfully developed a robust covalent surface functionalization technique for GO-based fluorescent aptasensors, demonstrating improved selectivity and resistance to nonspecific probe displacement compared to physisorbed aptasensors. The method is versatile and can be adapted for different analytical applications by changing the aptamer sequence.
研究不足
The study acknowledges the technical constraints of physisorbed aptasensors, such as poor signal reversibility and reproducibility, and nonspecific probe displacement. The covalent aptasensor developed addresses these limitations but may still require optimization for specific applications.
1：Experimental Design and Method Selection:
The study involved the covalent functionalization of graphene oxide (GO) with an amino-silane and glutaraldehyde to create available carbonyl groups for the covalent attachment of a fluorophore and an amino dual modified ATP binding aptamer (ABA). The methodology included surface characterization techniques like zeta potential, XPS, and FTIR to monitor each step of the GO surface functionalization.
2：Sample Selection and Data Sources:
The samples used included graphene oxide, amino-silane, glutaraldehyde, and a well-characterized ATP binding aptamer (ABA) labeled with FAM. Data sources included fluorescence measurements, zeta potential measurements, XPS, and FTIR spectra.
3：List of Experimental Equipment and Materials:
Equipment included a Malvern Zetasizer for zeta potential measurements, X-ray photoelectron spectrometer (XPS), Fourier transform infrared spectrometer (FTIR), and a microplate reader for fluorescence measurements. Materials included graphene oxide, amino-silane, glutaraldehyde, ATP binding aptamer, and various buffers.
4：Experimental Procedures and Operational Workflow:
The experimental procedure involved the modification of GO with amino-silane, followed by reaction with glutaraldehyde to create carbonyl groups for aptamer attachment. The aptamer was then covalently attached to the modified GO, and the resulting aptasensor was characterized for performance.
5：Data Analysis Methods:
Data analysis included fitting zeta potential data to the Smoluchowski model, analyzing XPS and FTIR spectra for chemical composition and bond variations, and using statistical analysis software OriginPro for statistical evaluation of experimental data.
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