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Localized Visualization and Autonomous Detection of Cell Surface Receptor Clusters Using DNA Proximity Circuit
Yan Shan Ang1, Jia'En Jasmine Li1, Pei-Jou Chua2
1Department of Chemical and Biomolecular Engineering , National University of Singapore , Singapore 117585 , Singapore.
Analytical Chemistry
|April 3, 2018
Summary
We developed a split proximity circuit (SPC) for detecting cell surface receptor clusters. This enzyme-free method visualizes human epidermal growth factor receptor (HER) family dimers in cells and lysates.
Area of Science:
- Biotechnology and Molecular Biology
- Cellular Signaling and Receptor Biology
Background:
- Cell surface receptors are crucial for cell communication, disease biomarker identification, and therapeutic targeting.
- Accurate detection and visualization of cell surface receptor clusters are essential for understanding cellular processes and disease states.
Purpose of the Study:
- To introduce a novel one-pot molecular toolbox, the split proximity circuit (SPC).
- To enable autonomous detection and visualization of cell surface receptor clusters.
- To apply the SPC for studying the human epidermal growth factor receptor (HER) family.
Main Methods:
- Development of a split proximity circuit (SPC) utilizing antibody recognition and DNA hybridization for signal amplification.
- Enzyme-free, localized signal amplification strategy for enhanced detection sensitivity.
- Application of SPC for detecting HER2:HER2 homodimers and HER2:HER3 heterodimers in cell lysate and fixed whole cells.
- Detection and imaging using standard microplate scans and confocal microscopy.
Main Results:
- The SPC system demonstrated specific detection with minimal background leakage.
- Successful visualization and profiling of HER family receptor expression in three distinct cell types.
- The method operated efficiently in a one-pot format, eliminating the need for intermediate washing steps.
Conclusions:
- The split proximity circuit (SPC) offers a robust and efficient platform for the autonomous detection and visualization of cell surface receptor clusters.
- The SPC system is suitable for analyzing receptor expression profiles in various biological contexts, including cell lysates and intact cells.
- This technology holds promise for advancing research in cell communication, biomarker discovery, and targeted therapies.
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