Reduced graphene oxide decorated with gold nanoparticle as signal amplification element on ultra-sensitive electrochemiluminescence determination of caspase-3 activity and apoptosis using peptide based biosensor

还原氧化石墨烯修饰金纳米粒子作为信号放大元件,用于肽基生物传感器超灵敏电化学发光测定 caspase-3 活性和细胞凋亡

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作者:Balal Khalilzadeh, Nasrin Shadjou, Hadi Afsharan, Morteza Eskandani, Hojjatollah Nozad Charoudeh, Mohammad-Reza Rashidi

Conclusion

The prepared peptide based biosensor could be considered as an excellent candidate for early detection of apoptosis, cell turnover, and cancer related diseases.

Methods

The ECL intensity of the luminol was improved by using the streptavidin coated magnetic beads and HRP in the presence of hydrogen peroxide. The cleavage behavior of caspase-3 was characterized by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) techniques using biotinylated peptide (DEVD containing peptide) which was coated on reduced graphene oxide decorated with gold nanoparticle. The surface modification of graphene oxide was successfully confirmed by FTIR, UV-vis and x-ray spectroscopy.

Results

ECL based biosensor showed that the linear dynamic range (LDR) and the lower limit of quantification (LLOQ) were 0.5-100 and 0.5 femtomolar (fM), respectively. Finally, the performance of the engineered peptide based biosensor was validated in the A549 cell line as real samples.

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