Protein-Templated Click Ligation Reaction Triggered by Protein-Split Aptamer Interactions.

Area of Science:

• Biochemistry
• Molecular Biology
• Biosensing Technology

Background:

• DNA-templated reactions are valuable in sensing and drug discovery.
• Current methods are limited to nucleic acid templates, restricting broader applications.
• Protein-templated reactions offer a novel approach to proximity-directed chemistry.

Purpose of the Study:

• To introduce a versatile protein-templated split aptamer click ligation reaction (PT-SpA-CLR).
• To demonstrate the translation of protein-aptamer binding into ligated DNA products.
• To develop sensitive and low-cost biosensors for protein biomarker detection.

Main Methods:

• Utilized split aptamer elements that ligate upon protein binding to a protein template.
• Developed a protein-templated split aptamer click ligation reaction (PT-SpA-CLR).
• Integrated PT-SpA-CLR with colorimetric and fluorescent reporting mechanisms for biosensing.

Main Results:

• Achieved >80% ligation yield for three model protein templates (VEGF165, PDGF-BB, SARS-CoV-2 S1).
• The ligation strategy improved binding affinity, showing a 2-fold lower dissociation constant.
• Developed biosensors with picomolar limit of detection, functional in biological samples.

Conclusions:

• PT-SpA-CLR is a versatile strategy for translating protein binding into DNA ligation.
• The developed biosensors are sensitive, specific, low-cost, and easy to use.
• This platform offers a universal approach for efficient protein biomarker detection.