Journal
BIOELECTROCHEMISTRY
Volume 145, Issue -, Pages -Publisher
ELSEVIER SCIENCE SA
DOI: 10.1016/j.bioelechem.2022.108090
Keywords
Caspase-3; Apoptosis; Electrochemical phage sensor; Chemical modification
Funding
- National Research Foundation of Korea (NRF) - Korean government (MSIT) [2019R1A2C2084065, 2021R1A4A1022206]
- National Research Foundation of Korea [2021R1A4A1022206, 2019R1A2C2084065] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
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In this study, a phage-based electrochemical biosensor was developed for the evaluation of cell apoptosis by detecting caspase-3. The sensor showed good binding affinity and detection limit for caspase-3 detection, and could be used for the diagnosis and prognosis of apoptosis-related diseases.
Caspase-3, a cysteine-dependent protease, is considered a reliable molecular biomarker for the diagnosis and prognosis of apoptosis-related diseases. In this study, we demonstrated a phage-based electrochemical biosensor for the evaluation of cell apoptosis by the sensitive and specific detection of caspase-3. Specifically, for screening of affinity peptide-displayed phages, phage display was performed using M13 phage libraries (cyclic forms of peptides), and we identified potential affinity peptide-displayed phage clones with the sequence CPTTMWRYC. After characterization of its binding affinity using enzyme-linked immunosorbent assay, whole phage particles were covalently attached to a gold surface using coupling chemistry (MUA-EDC/NHS). The developed phage sensor was characterized by X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), scanning electron microscopy (SEM), electrochemical analysis using cyclic voltammetry (CV), and square wave voltammetry (SWV). Under optimal conditions, the affinity peptide-displayed phage sensor showed a good binding affinity (K-d = 0.13 +/- 0.56 mu M) and limit of detection (0.39 mu M) for caspase-3 detection. Furthermore, developed phage sensor could be monitored the response of apoptotic HeLa cells by detecting caspase-3 activity. This work should stimulate the development of efficient alternative caspase-3 detection methods for the diagnosis and prognosis of apoptosis-related diseases.
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