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Immobilization coupling with aptamer assisted dual cycle amplification for sensitive sEVs isolation and analysis
Yu He1, Ying Ren2, Jiawen Tang2
1Department of Pathology, The Affiliated People's Hospital of Ningbo University, No. 251 Baizhang Road, Yinzhou District, Ningbo, 315040, Zhejiang Province, China. lili15036@sina.com.
Biotechnology Letters
|September 12, 2024
Summary
This study presents a novel aptamer-assisted dual cycle amplification technique for sensitive isolation and analysis of small extracellular vesicles (sEVs). The method enables precise quantification and tracing of sEVs, crucial for disease diagnosis and personalized medicine.
Area of Science:
- Biotechnology
- Nanotechnology
- Biomedical Engineering
Background:
- Precise identification and quantification of small extracellular vesicles (sEVs) are critical for advancing disease diagnosis and therapeutic strategies, particularly in conditions like bladder cancer.
- Current methods for sEV isolation and analysis face challenges in achieving high sensitivity and simultaneous quantification.
- Developing robust techniques for sEV characterization is essential for their clinical translation.
Purpose of the Study:
- To introduce a novel technique combining aptamer-assisted dual cycle amplification with immobilization for highly sensitive isolation and analysis of sEVs.
- To enable simultaneous quantification and tracing of sEVs through dual biomarker analysis.
- To establish a stable and accurate method for sEV detection with a broad dynamic range and low limit of detection.
Main Methods:
- Immobilization of CD9 protein antibody on a plate surface for initial sEV capture.
- Aptamer probe-based detection of the exosomal surface protein CD63.
- Integration of target recognition-initiated signal recycling and rolling circle amplification (RCA) for signal amplification.
- Implementation of "AND" logic for dual biomarker analysis.
Main Results:
- The developed method demonstrates high sensitivity in isolating and analyzing sEVs.
- Achieved a broad detection range and a low limit of detection for sEVs.
- Exhibited good stability with a low coefficient of variation in sEV detection.
- Successfully enabled simultaneous quantification and tracing of sEVs.
Conclusions:
- The proposed aptamer-assisted dual cycle amplification method offers a significant advancement in sEV isolation and identification.
- This technique provides accurate quantification and tracing capabilities, suitable for various applications in biological science and personalized medicine.
- The method's sensitivity, stability, and broad detection range make it a promising tool for clinical diagnostics and research.
