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Optimized Functional Covalent Immobilization of DNA on Plasma-Activated Surfaces for Biosensing Applications
Kanako Coffi Dit Gleize1, Bradley I Harding1,2, Qianyi Zhang1,2
1School of Chemistry, The University of Sydney, Sydney, New South Wales 2006, Australia.
We developed a simple, high-yield method for immobilizing DNA probes on surfaces using plasma-activated coating and click chemistry. This improves DNA surface density and enables stable, reusable biosensors for diverse applications.
Area of Science:
- Biophysics
- Biosensing Technologies
- Nanotechnology
Background:
- Surface-immobilized DNA is crucial for biosensors, aptamers, and DNA nanostructures.
- Current methods for DNA surface immobilization are often toxic, slow, and limited by DNA's negative charge and orientation.
- Functional DNA biosensors require precise control over DNA orientation on surfaces.
Purpose of the Study:
- To develop an improved, high-yield method for DNA surface immobilization.
- To enhance DNA immobilization density and stability on surfaces.
- To demonstrate the utility of this method in creating functional biosensors.
Main Methods:
- Utilized plasma-activated coating (PAC) on surfaces.
- Modified DNA probes with a glycine-polyethylene glycol (Gly-PEG) linker via click chemistry.
- Assessed DNA immobilization yield, stability, and binding capabilities.
Main Results:
- Achieved a 3-fold increase in DNA immobilization on PAC-treated plates using Gly-PEG modification.
- Demonstrated stable immobilization allowing multiple hybridization/dehybridization cycles.
- Showcased binding of various targets, including DNA origami nanostructures.
- Confirmed improved biosensor performance due to controlled DNA orientation.
Conclusions:
- The PAC and Gly-PEG modification method offers a simple, high-yield approach for DNA surface immobilization.
- This technique enhances biosensor stability, reusability, and performance.
- Enables advanced applications in biosensing, biophysics, and complex surface patterning with DNA nanostructures.
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