iSECRETE: Integrating Microfluidics and DNA Proximity Amplification for Synchronous Single-Cell Activation and IFN-γ...

Area of Science:

• Immunology
• Biotechnology
• Cellular Biology

Background:

• Cytokine dysregulation significantly impacts immune modulation and disease progression.
• Current cytokine profiling methods are often slow, labor-intensive, and fail to capture dynamic immune responses.

Purpose of the Study:

• To develop an integrated platform, iSECRETE, for rapid, single-cell cytokine secretion analysis.
• To enable synchronous cell activation and target-responsive protein detection within a short timeframe.

Main Methods:

• Utilized a multifunctional microfluidic system incorporating a DNA proximity assay (DPA).
• Achieved one-pot homogenous cytokine signal amplification for sensitive detection.
• Demonstrated wash-free, target-responsive protein detection at room temperature.

Main Results:

• iSECRETE provides single-cell IFN-γ cytokine secretion analysis within 30 minutes.
• Achieved a limit of detection of approximately 50 secreted molecules per cell.
• Successfully detected single-cell IFN-γ secretion from CAR T cells upon cancer target exposure within 6 hours.

Conclusions:

• iSECRETE is the fastest single-cell sample-to-result cytokine secretion assay to date.
• The platform offers a powerful tool for understanding biological phenotypes, functions, and pathways under in vivo-like conditions.
• Enables robust analysis across various sample types.