Nanomaterial-based electrochemical biosensors for cytochrome c using cytochrome c reductase

Pandiaraj, Manickam; Madasamy, Thangamuthu; Naidu Gollavilli, Paradesi; Balamurugan, Murugesan; Kotamraju, Srigiridhar; Kameswara Rao, Vepa; Bhargava, Kalpana; Karunakaran, Chandran

Abstract

Emerging evidences have pointed out that the release of cytochrome c (cyt c) from mitochondria into cytosol is a critical step in the activation of apoptosis. This article presents a novel approach for the detection of mitochondrial cyt c release for the first time using cytochrome c reductase (CcR) immobilized on nanoparticles decorated electrodes. Two kinds of nanomaterial-based biosensor platforms were used: (a) carbon nanotubes (CNT) incorporated polypyrrole (PPy) matrix on Pt electrode and (b) self-assembled monolayer (SAM) functionalized gold nanoparticles (GNP) in PPy-Pt. Scanning electron microscope was used to characterize the surface morphologies of the nanomaterial modified electrodes. Cyclic voltammograms of both the biosensors showed reversible redox peaks at − 0.45 and − 0.34 V vs Ag/AgCl, characteristic of CcR. In comparison, the CcR-CNT biosensor gave a detection limit of 0.5 ± 0.03 μM cyt c, which was 4-fold better than the CcR-GNP biosensor (2 ± 0.03 μM). Moreover, the CcR-CNT biosensor achieved a much larger linear range (1–1000 μM) over the CcR-GNP biosensor (5–600 μM) with 2-fold better sensitivity. The CcR-CNT-PPy-Pt biosensor was further applied to quantify the mitochondrial cyt c released in cytosol of A549 cells upon induction of apoptosis with doxorubicin, the results agreed well with standard western blot analysis.

Citation

Pandiaraj, M., Madasamy, T., Naidu Gollavilli, P., Balamurugan, M., Kotamraju, S., Kameswara Rao, V., Bhargava, K., & Karunakaran, C. (2013). Nanomaterial-based electrochemical biosensors for cytochrome c using cytochrome c reductase. Bioelectrochemistry, 91, 1-7. https://doi.org/10.1016/j.bioelechem.2012.09.004

• Journal Article Type: Article
• Acceptance Date: Sep 29, 2012
• Online Publication Date: Nov 6, 2012
• Publication Date: 2013-06
• Deposit Date: Aug 1, 2023
• Journal: Bioelectrochemistry
• Print ISSN: 1567-5394
• Publisher: Elsevier
• Peer Reviewed: Peer Reviewed
• Volume: 91
• Pages: 1-7
• DOI: https://doi.org/10.1016/j.bioelechem.2012.09.004
• Public URL: https://nottingham-repository.worktribe.com/output/23549875
• Publisher URL: https://www.sciencedirect.com/science/article/pii/S156753941200165X