Sensitive and specific detection of proteins based on target-responsive DNA polymerase activity

Area of Science:

• Biotechnology
• Molecular Biology
• Analytical Chemistry

Background:

• Protein detection is crucial in diagnostics and research.
• Existing methods may have limitations in sensitivity or complexity.
• Developing novel, sensitive, and efficient protein detection systems is essential.

Purpose of the Study:

• To develop a novel protein detection method utilizing target-responsive DNA polymerase activity.
• To engineer a detection probe combining two DNA aptamers for target protein sensing.
• To achieve highly sensitive and specific detection of target proteins.

Main Methods:

• Designed a sensor probe with two DNA aptamers: one for target protein binding, one for DNA polymerase binding.
• Utilized target protein presence to destabilize the probe, releasing inhibition of DNA polymerase.
• Employed DNA polymerase-catalyzed primer extension and TaqMan probe for signal amplification.

Main Results:

• Achieved sensitive detection of a model target protein, lysozyme, with a limit of detection as low as 0.80 nM.
• Demonstrated the practical applicability of the system by successfully determining lysozyme in human serum.
• The assay showed target-responsive DNA polymerase activation and signal amplification.

Conclusions:

• The developed method offers a novel and sensitive approach for protein detection.
• The target-responsive DNA polymerase system shows promise for diagnostic applications.
• This strategy provides a robust platform for detecting various target proteins.