FRET-Based Aptasensor for the Selective and Sensitive Detection of Lysozyme

Area of Science:

• Biochemistry
• Molecular Biology
• Biotechnology

Background:

• Lysozyme is a key antimicrobial enzyme involved in immune modulation.
• Elevated lysozyme levels in body fluids can indicate diseases like Alzheimer's, Crohn's, and breast cancer.
• Existing lysozyme detection methods lack sensitivity, specificity, and quantitative capabilities.

Purpose of the Study:

• To develop a sensitive, selective, and quantitative detection method for lysozyme.
• To utilize aptamers for lysozyme detection via a single-molecule fluorescence resonance energy transfer (smFRET) approach.
• To create a cost-effective and regenerable aptasensor for protein biomarker detection.

Main Methods:

• Development of an aptasensor utilizing aptamers specific to lysozyme.
• Implementation of single-molecule fluorescence resonance energy transfer (smFRET) to detect lysozyme binding.
• Employing toehold mediated strand displacement (TMSD) for aptasensor regeneration.

Main Results:

• The aptasensor demonstrated high sensitivity, detecting lysozyme down to 2.3 picomoles (30 nM) with a dynamic range up to ~2 µM.
• The method showed minimal interference from similar biomolecules, ensuring specificity.
• The smFRET approach required significantly less aptasensor material compared to bulk methods and was cost-effective.

Conclusions:

• The developed smFRET-based aptasensor provides a sensitive, selective, and quantitative method for lysozyme detection.
• This aptasensing strategy is cost-effective, requires minimal reagents, and is regenerable.
• The platform is adaptable for detecting other protein biomarkers by simply changing the protein-specific aptamer.