Aptamer-based biosensor arrays for detection and quantification of biological macromolecules

Area of Science:

• Biotechnology
• Molecular Diagnostics
• Biosensor Technology

Background:

• Multiplex analysis of protein biomarkers is crucial for accurate disease diagnosis.
• Existing methods for protein detection can be complex and time-consuming.
• Aptamers offer a promising alternative to antibodies for molecular recognition.

Purpose of the Study:

• To develop and validate a chip-based biosensor for simultaneous detection of multiple protein analytes.
• To assess the utility of immobilized DNA and RNA aptamers for protein quantification.
• To demonstrate the biosensor's capability in detecting cancer-associated proteins in biological samples.

Main Methods:

• Development of a chip-based biosensor platform utilizing immobilized, fluorescently labeled DNA and RNA aptamers.
• Selection of aptamers against specific protein targets.
• Application of fluorescence polarization anisotropy for solid- and solution-phase binding measurements.
• Testing the biosensor with cancer-associated proteins in human serum and cellular extracts.

Main Results:

• Demonstrated that solid-phase aptamer-protein interactions correlate with solution-phase binding.
• Achieved specific detection and quantification of multiple cancer-associated proteins, including inosine monophosphate dehydrogenase II, vascular endothelial growth factor, and basic fibroblast growth factor.
• Successfully validated the biosensor's performance in complex biological matrices like human serum and cellular extracts.

Conclusions:

• The developed chip-based aptamer biosensor enables multiplexed protein analysis with high specificity.
• This technology can accurately detect and quantify cancer-associated proteins in relevant biological samples.
• The biosensor has the potential to accelerate cancer diagnosis by enabling rapid, direct protein expression analysis.