Discover a new picodroplet-based approach for the functional validation of CAR-T cells.

Explore the revolutionary method for studying cell-mediated cytotoxicity using picodroplet technology.

This application note introduces an easy-to-use, robust technique for the functional validation of CAR-T cells, co-encapsulating engineered T cells and target cells in picolitre-volume aqueous droplets (picodroplets), along with granzyme B substrate.

By capturing granzyme B release, a readout of the T cell’s ability to recognize and kill cancer cells, this method provides rapid cell product efficacy readouts, streamlining the QC release of cell products.

Introduction to Picodroplet Technology

In recent years, cell therapy has gained significant traction as a method to harness the immune system in combating tumors. One key technology involves reprogramming T cells from patients to create personalized medicines, restoring their ability to attack cancer cells.

However, accurate assays to qualify the potency of these engineered T-cells are critical for developing effective therapeutics. Our new picodroplet-based approach integrates high-throughput picodroplet technology with a granzyme B assay, offering a superior method for studying cell-mediated cytotoxicity.

How It Works

Engineered T cells and target cells are co-encapsulated in picolitre-volume aqueous droplets (picodroplets) within an oil emulsion, along with granzyme B substrate. The release of granzyme B by the T cell, indicative of its ability to recognize and kill cancer cells, is captured, determining the level of potential anti-cancer activity. This process enables rapid efficacy readouts, making it an invaluable tool for quality control in cell product release.

Benefits of Picodroplet Technology

Picodroplets provide a unique microenvironment for single-cell functional analysis, offering several advantages over traditional methods:

• High Throughput: Allows for rapid analysis of large cell populations.
• Increased Sensitivity: Miniaturized volumes reduce sample dilution and increase detection sensitivity.
• Efficiency: Reduces sample requirements and reaction times, enhancing overall efficiency.

Detailed Workflow and Granzyme B Detection

The granzyme B detection assay utilizes a commercially available kit adapted for use in picodroplets. This assay is based on the cleavage of a granzyme B substrate peptide, labeled with a 5-FAM fluorophore and a QXL®-520 quencher. When granzyme B cleaves the substrate, the quenching molecule is released, emitting a fluorescent signal that indicates cytotoxic activity.

Case Study: CAR-T and Target Cells

In our study, we co-encapsulated CAR-T cells expressing a CAR against Prostate-Specific Membrane Antigen (PSMA) with target cells expressing PSMA. Fluorescent signals were monitored over time, demonstrating the efficacy of granzyme B release in detecting CAR-T cell activity. This method provides a rapid, accurate assessment of CAR-T cell potency, crucial for therapeutic development.

Cyto-Mine® Integration

Our Cyto-Mine® Single Cell Analysis System enhances this process by automating detection and sorting of granzyme B positive picodroplets. This integration further streamlines the workflow, allowing for high-throughput, precise analysis and sorting of cell populations.

Conclusion

This picodroplet-based approach for CAR-T cell function verification offers a high-throughput, sensitive, and efficient method for cytotoxicity analysis. By providing rapid efficacy readouts, it supports the development of potent, personalized cell therapies. Explore how our innovative technology can transform your lab workflow and accelerate your research in drug discovery and cell therapy development.

Download the Full Application Note

Unlock the full potential of picodroplet technology in CAR-T cell function verification. Download our comprehensive application note to learn more about this groundbreaking approach and how it can enhance your research and therapeutic development processes: