Detection of Early Stage Apoptotic Cells Based on Label-Free Cytochrome c Assay Using Bioconjugated Metal Nanoclusters as Fluorescent Probes

Analytical Chemistry
Mojtaba ShamsipurFereshteh Rahmati

Abstract

Cytochrome c (Cyt c) is an important biomarker in cell lysates for the early stage of apoptosis or anticancer agents. Here, two novel label-free fluorescence assays based on hemoglobin-stabilized gold nanoclusters (Hb/AuNCs) and aptamer-stabilized silver nanoclusters (DNA/AgNCs) for analysis of Cyt c are presented. The heme group of the protein induces sensitive sensing platforms accompanied by the decreased fluorescence of both metal nanoclusters. The quenching processes observed found to be based on the fluorescence resonance energy transfer mechanism from Hb/AuNCs to Cyt c and photoinduced electron transfer from DNA/AgNCs to the aptamer-Cyt c complex. The linear range for Cyt c was found to be 0-10 μM for Hb/AuNCs and from 0 to 1 μM for DNA/AgNCs, with limits of detection of ∼15 nM. On the basis of strong binding affinity of DNA aptamers for their target proteins, the DNA/AgNCs probe was successfully applied to the quantitative determination of Cyt c in cell lysates, which opens a new avenue to early diagnostics and drug screening with high sensitivity. Compared to the conventional Western blot method, the presented assays are low cost, easy to prepare the fluorescent probes, and sensitive, while overall time for the detecti...Continue Reading

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Citations

Sep 14, 2016·Biosensors & Bioelectronics·Pandiaraj ManickamShekhar Bhansali
Sep 16, 2016·Journal of Biomolecular Structure & Dynamics·Vida Jafari AzadMojtaba Falahati
Nov 30, 2016·Biosensors & Bioelectronics·Mojtaba ShamsipurSaman Hosseinkhani
Oct 21, 2017·Journal of Materials Chemistry. B, Materials for Biology and Medicine·Sebastián A DíazEunkeu Oh
Aug 5, 2017·Materials·Marina AlloisioDavide Comoretto
Jul 21, 2018·Journal of Materials Chemistry. B, Materials for Biology and Medicine·Mingzhu ZhangYupeng Tian
Oct 24, 2020·Mikrochimica Acta·Mohsen AlipourSaman Hosseinkhani
Apr 17, 2020·Analytical Chemistry·Mei LiJian-Feng Li
Oct 14, 2017·Chemical Reviews·Damián Alvarez-PaggiDaniel H Murgida

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Apoptosis

Apoptosis is a specific process that leads to programmed cell death through the activation of an evolutionary conserved intracellular pathway leading to pathognomic cellular changes distinct from cellular necrosis