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Bacterial Detection & Identification Using Electrochemical Sensors
Published on: April 23, 2013
Detecting multiple cell-secreted cytokines from the same aptamer-functionalized electrode
Ying Liu1, Ying Liu1, Zimple Matharu1
1Department of Biomedical Engineering, University of California, Davis, Davis, CA 95616, USA.
Insights
This study developed an electrochemical aptasensor for simultaneous detection of interferon gamma (IFN-γ) and tumor necrosis factor alpha (TNF-α). The sensor enables dynamic monitoring of cytokine release from immune cells for disease diagnosis.
Area of Science:
- Biomedical Engineering
- Analytical Chemistry
- Immunology
Background:
- Inflammatory cytokines like interferon gamma (IFN-γ) and tumor necrosis factor alpha (TNF-α) are crucial biomarkers for disease diagnosis.
- Current methods for cytokine quantification can be limited in parallel detection capabilities.
Purpose of the Study:
- To develop an electrochemical aptasensor for the simultaneous detection of IFN-γ and TNF-α.
- To enable multiplexed cytokine analysis for illuminating inflammatory pathways.
Main Methods:
- Development of an electrochemical aptasensor using aptamers labeled with unique redox reporters (anthraquinone and methylene blue).
- Random immobilization of aptamers on a gold electrode for distinct redox peak detection.
- Integration of the aptasensor into microfluidic devices for dynamic monitoring of cytokine release from immune cells.
Main Results:
- The aptasensor demonstrated sensitive and specific detection of IFN-γ and TNF-α through changes in redox signals.
- Simultaneous monitoring of IFN-γ and TNF-α release from primary human CD4 T-cells and U937 monocytic cells was achieved over 2 hours.
- The strategy allows for multiplexed protein biomarker detection from a single electrode.
Conclusions:
- The developed electrochemical aptasensor offers a sensitive and specific platform for simultaneous quantification of multiple inflammatory cytokines.
- This technology has potential applications in disease diagnosis by analyzing inflammatory pathways and monitoring immune cell responses.
Abstract:
Inflammatory cytokines are secreted by immune cells in response to infection or injury. Quantification of multiple cytokines in parallel may help with disease diagnosis by illuminating inflammatory pathways related to disease onset and progression. This paper describes development of an electrochemical aptasensor for simultaneous detection of two important inflammatory cytokines, interferon gamma (IFN-γ) and tumor necrosis factor alpha (TNF-α). To enable multiplexing, IFN-γ and TNF-α aptamers were labeled with anthraquinone (AQ) and methylene blue (MB) redox reporters respectively. Random immobilization of two aptamer on gold exhibited redox peaks at -0.37 V (AQ) and -0.15 V (MB) vs. Ag/AgCl reference. When challenged with either IFN-γ or TNF-α, redox signal of the appropriate reporter changed in concentration dependent manner. To demonstrate one possible application of this sensing approach, electrodes were integrated into microfluidic devices and used to dynamically monitor cytokine release from immune cells. Two cell types, primary human CD4 T-cells and U937 monocytic cells, were used to compare differences in cytokine secretions upon stimulation. These cells were infused into the microfluidic devices and stimulated to commence cytokine production. Release of IFN-γ and TNF-α was monitored concurrently from the same small group of cells over the course of 2h. The strategy of encoding specific aptamer types with unique redox reporters allows sensitive and specific detection of multiple protein biomarkers from the same electrode.
