Functionalized DNA-spider silk nanohydrogels for controlled protein binding and release.

Area of Science:

• Biomaterials Science
• Biotechnology
• Biochemistry

Background:

• Hydrogels offer protective enzyme scaffolds but lack selective immobilization methods for bioanalytical devices.
• Current techniques struggle with substrate attachment and precise biocatalyst anchoring.
• Recombinant spider silk proteins (rssp) present a novel material for nanohydrogel formation.

Purpose of the Study:

• To engineer spider silk nanohydrogels with bioselective protein-binding capabilities.
• To create a functionalized scaffold for sensitive enzyme immobilization and controlled release.
• To demonstrate the utility of aptamer-functionalized nanohydrogels in bioanalytical applications.

Main Methods:

• Fabrication of nanofilm coatings using recombinant spider silk protein (rssp) for self-assembly into fibril-based nanohydrogels.
• Chemical conjugation of antithrombin aptamers to the rssp nanohydrogel network for selective protein binding.
• Utilizing human thrombin as a model target enzyme, assessing its activity and release.
• Employing complementary DNA sequences to trigger aptamer structural changes and induce enzyme release.

Main Results:

• Successfully created fibril-based nanohydrogels from rssp with integrated aptamer functionality.
• Demonstrated selective and cooperative embedding of human thrombin into the nanohydrogels via aptamer binding.
• Confirmed the release of active thrombin upon structural alteration of the aptamers using DNA sequences.
• Validated the addressable functionalization of spider silk nanohydrogels for controlled enzyme release.

Conclusions:

• Spider silk nanohydrogels can be functionalized with aptamers for selective enzyme capture and release.
• This approach overcomes limitations of traditional hydrogel immobilization for bioanalytical devices.
• The developed system offers a promising platform for sensitive enzyme-based detection and assays.