From Gold to Polymer: Latex Bead-Assisted CRISPR-Cas12a Platform for Next-Generation Protein Diagnostics

Area of Science:

• Biomedical Engineering
• Molecular Diagnostics
• Biotechnology

Background:

• Sensitive detection of protein biomarkers like interleukin-6 (IL-6) is vital for disease diagnosis and treatment monitoring.
• Existing gold nanoparticle (AuNP)-assisted CRISPR-Cas12a biosensors face challenges including high cost, aggregation, and nonspecific adsorption.
• There is a need for improved biosensing platforms for clinical applications.

Purpose of the Study:

• To develop a CRISPR-Cas12a-based fluorescence biosensor for detecting the inflammatory biomarker IL-6.
• To replace gold nanoparticles with latex beads as transducers and signal amplifiers in the biosensor.
• To evaluate the sensitivity, selectivity, and practical applicability of the novel biosensor.

Main Methods:

• Utilized a CRISPR-Cas12a system with dual-functionalized latex beads carrying IL-6 detection antibodies and dsDNA strands.
• Employed IL-6 antibody-functionalized magnetic beads for sample preparation and enrichment.
• Integrated latex beads as signal amplifiers for fluorescence detection.

Main Results:

• Achieved a highly sensitive limit of detection (LOD) for IL-6 as low as 0.76 pg/mL.
• Demonstrated excellent selectivity, with no interference from other proteins like HSA, BSA, CRP, PCT, and IL-2β.
• Successfully validated the biosensor's performance in analyzing simulated serum samples.

Conclusions:

• Latex beads are a viable and advantageous alternative to gold nanoparticles in CRISPR-based protein diagnostics.
• The developed biosensor offers improved stability and specificity while maintaining high sensitivity for IL-6 detection.
• This approach holds significant potential for early disease diagnosis and monitoring through enhanced protein biomarker analysis.