FRET-Based TURN-ON Aptasensor for the Sensitive Detection of CK-MB.

Area of Science:

• Biochemistry
• Analytical Chemistry
• Biotechnology

Background:

• Immunoassays typically use antibodies to detect biomarkers.
• Existing methods for quantifying cardiac marker CK-MB may lack sensitivity or require complex procedures.

Purpose of the Study:

• To develop a novel, highly sensitive fluorescence assay for quantifying CK-MB.
• To innovate an enzyme-linked aptamer assay (ELAA) by replacing antibodies with aptamers.

Main Methods:

• A sandwich assay was designed using biotin-labeled aptamers (apt. A1 and apt. A2) targeting CK-MB.
• Avidin-linked alkaline phosphatase (ALP) catalyzed substrate hydrolysis, producing ascorbic acid.
• Ascorbic acid reduced MoS2 nanosheets to nanoribbons, releasing quenched aggregation-induced emission quantum dots (AGQDs).
• Förster Resonance Energy Transfer (FRET) between nanoribbons and AGQDs reestablished fluorescence for detection.

Main Results:

• The developed assay demonstrated a working detection range of 2.5 nM to 160 nM for CK-MB.
• A low limit of detection (LOD) of 0.20 nM for CK-MB was achieved.
• The assay showed enhanced sensitivity for disease biomarker detection.

Conclusions:

• The enzyme-linked aptamer assay (ELAA) provides a sensitive and effective method for CK-MB quantification.
• Aptamer-based assays offer a promising alternative to traditional antibody-based immunoassays.
• This approach enhances disease biomarker detection sensitivity.