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Universal aptameric system for highly sensitive detection of protein based on structure-switching-triggered rolling
Zai-Sheng Wu1, Songbai Zhang, Hui Zhou
1State Key Laboratory for Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, P. R. China.
Analytical Chemistry
|February 16, 2010
Summary
This study introduces a universal aptamer assay for protein detection using structure-switching triggered ligation-rolling circle amplification (L-RCA). This novel method offers high sensitivity and specificity for detecting various proteins.
Area of Science:
- Biotechnology
- Molecular Biology
- Analytical Chemistry
Background:
- Aptamers offer specific molecular recognition for biosensing.
- Rolling circle amplification (RCA) provides a sensitive signal amplification method.
- Existing aptamer assays can lack universality and require specific optimization.
Purpose of the Study:
- To develop a universal aptamer-based assay system for protein detection.
- To leverage structure-switching aptamers and ligation-rolling circle amplification (L-RCA) for enhanced sensitivity.
- To create a versatile platform adaptable for detecting various protein targets.
Main Methods:
- Aptamer probe design for target-specific structure switching.
- Competition assay between target protein and complementary DNA (cDNA) for aptamer binding.
- Ligation-rolling circle amplification (L-RCA) triggered by target-free aptamer-cDNA complex.
- Signal detection via hybridization of RCA products with molecular beacons.
Main Results:
- Achieved attomolar detection limits for proteins.
- Demonstrated a linear dynamic range exceeding 3 orders of magnitude.
- Exhibited high specificity and sensitivity in protein detection.
Conclusions:
- The developed aptamer-L-RCA assay system is universal and adaptable for detecting diverse proteins.
- The assay offers significant advantages in sensitivity and specificity over existing methods.
- This platform provides a robust and versatile tool for protein analysis.
