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Related Experiment Video

Updated: Oct 22, 2025

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Aptamer-based cell-free detection system to detect target protein.

Junhong Chen1,2, Xiaoyan Zhuang1,2, Jiyang Zheng1,2

  • 1XMU-China Team, Xiamen University, Xiamen, 361005, PR China.

Synthetic and Systems Biotechnology
|September 1, 2021
PubMed
Summary

This study introduces an Aptamer-Based Cell-free Detection (ABCD) system for sensitive protein biomarker detection. The system utilizes aptamers and CRISPR-Cas12a for enhanced accuracy and a low limit of detection, paving the way for portable diagnostic devices.

Keywords:
AptamerBiomarkerCRISPR-CasProtein detection

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Biotechnology

Background:

  • Protein biomarkers are crucial for disease diagnosis and prognosis.
  • Antibody-based assays face limitations due to thermal instability and potential false positives in complex samples.
  • Aptamers offer a stable, versatile alternative to antibodies for molecular detection.

Purpose of the Study:

  • To develop a novel Aptamer-Based Cell-free Detection (ABCD) system for sensitive protein biomarker detection.
  • To leverage aptamer specificity and CRISPR-Cas12a signal amplification for improved assay performance.
  • To demonstrate the system's efficacy using epithelial cell adhesion molecule (EpCAM) as a target protein.

Main Methods:

  • Selection of aptamers using Systematic Evolution of Ligands by Exponential Enrichment (SELEX).
  • Integration of aptamers with CRISPR-Cas12a's trans-cleavage activity in a cell-free system.
  • Detection of epithelial cell adhesion molecule (EpCAM) using the developed ABCD system.

Main Results:

  • The ABCD system demonstrated robust detection of target proteins.
  • A low limit of detection (50-100 nM) was achieved for EpCAM.
  • The system showed superiority in detecting low-abundance proteins.

Conclusions:

  • The ABCD system offers a stable and sensitive platform for protein biomarker detection.
  • This technology provides a foundation for developing portable and advanced diagnostic devices.
  • The study highlights the potential of combining aptamers and CRISPR technology for future biosensing applications.