Proximity-dependent protein detection based on enzyme-assisted fluorescence signal amplification

Area of Science:

• Biochemistry
• Analytical Chemistry
• Molecular Biology

Background:

• Accurate and sensitive protein detection is crucial in diagnostics and research.
• Existing methods often face limitations in sensitivity or complexity.

Purpose of the Study:

• To develop a novel, highly sensitive fluorescence-based method for protein detection.
• To demonstrate the method's efficacy using specific protein targets.

Main Methods:

• Designing proximity probes with integrated recognition elements (aptamers or antibodies).
• Utilizing the proximity effect to convert protein binding events into DNA signals.
• Employing enzyme-assisted strand displacement for signal amplification.

Main Results:

• Successfully detected human α-thrombin and human IgG.
• Achieved a limit of detection (LOD) as low as 1 pM for human α-thrombin.
• Achieved a limit of detection (LOD) as low as 6 pM for human IgG.

Conclusions:

• The developed method offers high sensitivity and potential for broad protein detection.
• This fluorescence-based approach provides a sensitive platform for biomarker discovery and diagnostics.