Proteome Fishing for CRISPR/Cas12a-Based Orthogonal Multiplex Aptamer Sensing

Area of Science:

• Biotechnology
• Molecular Diagnostics
• Cancer Research

Background:

• Serum protein biomarker detection is complex due to serum's intricate nature.
• Existing methods face challenges in multiplexed protein amplification and aptamer stability in serum.

Purpose of the Study:

• To develop a novel platform for efficient and accurate detection of non-small cell lung cancer (NSCLC) biomarkers in serum.
• To establish a cost-effective and high-throughput diagnostic method for early cancer screening.

Main Methods:

• Utilized magnetic nanoparticle-protein corona for proteome fishing from serum.
• Developed a CRISPR/Cas12a-based orthogonal multiplex aptamer sensing (COMPASS) platform.
• Identified a four out of nine (FOON) panel (HE4, NSE, AFP, VEGF165) for NSCLC diagnosis.

Main Results:

• The FOON panel achieved high diagnostic accuracy for NSCLC: 95.56% (ROC-AUC = 99.40%) in internal cohorts and 89.58% (ROC-AUC = 95.41%) in external cohorts.
• The COMPASS technology successfully addressed challenges in multiplexed serum protein amplification and aptamer degradation.
• Demonstrated the feasibility of COMPASS for sensitive and specific biomarker detection.

Conclusions:

• The COMPASS platform provides a facile, cost-effective, and intelligent diagnostic solution for NSCLC.
• This technology has the potential for large-cohort cancer screening and early diagnosis.
• The developed method overcomes limitations of traditional serum biomarker detection techniques.